糖尿病复合营养酸奶降糖效应的实验研究
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摘要
目的:
糖尿病(DM)是一种全球范围内严重影响人类健康的常见内分泌代谢性疾病,其发病率和死亡率近年来呈逐步上升趋势。目前全世界已有糖尿病人近1.5亿,我国实际患病人数已达4000万人以上。DM主要是由于胰岛B细胞减少或B细胞功能减退引起的糖、脂肪、蛋白质、水及电解质代谢紊乱,并可由此产生各种并发症,如由大血管或微血管病变所引起的冠心病、脑血管疾病、视网膜病变、肾血管病变等,其中糖尿病患者有50%以上死于冠心病。因此,有效的改善糖尿病的代谢紊乱对预防糖尿病并发症、促进疾病康复和提高病人生活质量都具有十分重要的意义,而努力探索和积极寻找治疗和控制糖尿病的理想方法是医学界关注的重要课题。本研究以生物发酵作为切入点,通过观察我们所研制的糖尿病复合营养酸奶对保护糖尿病大鼠的胰腺功能、改善糖代谢、降低血糖血脂及对胰腺组织形态学、脂质过氧化状态和胰岛B细胞凋亡的影响,为糖尿病患者功效食品的研制提供实验依据。
方法:
清洁级成年Wister大鼠(雌雄各半)120只,体重190±10克,随机分为正常对照组、糖尿病对照组、治疗组、预防治疗组四组,每组30只。在常规饲料自由进食基础上,正常对照组给予生理盐水灌喂,糖尿病对照组和治疗组给予普通酸奶灌喂,预防治疗组给予糖尿病复合营养酸奶灌喂,隔日一次,每次2毫升,喂养至第15天按四氧嘧啶(ALX)50mg/k.w尾静脉注射制作糖尿病模型,正常对照组注射生理盐水。5天后测其空腹4小时后血糖,将血糖值>13.8mmol/L的大鼠做为糖尿病模型大鼠。糖尿病模型制作成功后除治疗组动物灌喂液由原来普通酸奶改为糖尿病复合营养酸奶外,其他组动物喂养方式不变,继续分组喂养,并于注射ALX后第30天、第90天再次进行空腹4小时血糖测定,注射ALX后第6天、第32天、第92天进行糖耐量测定,注射ALX后第33天、第93天分别从各组大鼠中随机取10只大鼠进行腹主动脉取血、处死大鼠取材进行以下指标测定:血清中的胰岛素(INS)、C-肽(C-P)、丙二醛(MDA)、超氧化物歧化酶(SOD)、肌酐(Cr)、尿素氮(BUN)、甘油三酯(TG)、总胆固醇(TC)和低密度脂蛋白(LDL)的含量,血浆中胰高血糖素(GLU)和糖化血红蛋白(GHb)含量,光镜下观察病理结构改变,TUNEL法测定胰岛B细胞凋亡,免疫组化法观察INS和GLU分泌的表达。
结果:
1.大鼠空腹血糖、糖耐量及糖化血红蛋白测定结果:
1.1 FBS水平(mmol/L):
1.1.1注射ALX后第5天,糖尿病对照组、治疗组、预防治疗组FBS非常显著高于正常对照组(P<0.01),预防治疗组非常显著低于糖尿病对照组和治疗组(P<0.01)。
1.1.2注射ALX后第30天和90天,糖尿病对照组、治疗组、预防治疗组非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组非常显著低于糖尿病对照组(P<0.01)。
1.2糖耐量测定结果(mmol/L):注射ALX后第6天、第32天、第92天糖尿病对照组、治疗组、预防治疗组动物0.5h、1.0h、1.5h、2.0h血糖测定值显著或非常显著高于正常对照组(P<0.05或p<0.01),但治疗组、预防治疗组第32天、92天上述各时相点血糖测定值非常显著低于糖尿病对照组(P<0.01),并且预防治疗组第6天上述各时相点血糖测定值非常显著低于治疗组(P<0.01)。
1.3糖化血红蛋白测定结果(%):注射ALX后第93天,糖尿病对照组、治疗组、预防治疗组糖化血红蛋白含量非常显著高于正常对照组(P<0.01),但治疗组和预防治疗组显著低于糖尿病对照组(P<0.01)。
2.大鼠血清INS、C-P、GLU测定结果:
2.1血清INS含量(μIU/ml):注射ALX后第33天、第93天,糖尿病对照组、治疗组、预防治疗组INS含量非常显著低于正常对照组(P<0.01),但治疗组、预防治疗组显著或非常显著高于糖尿病对照组(P<0.05或p<0.01)。
2.2血清C-P含量(ng/ml):注射ALX后第33天和93天,正常组、治疗组、预防治疗组C-P含量显著或非常显著高于糖尿病对照组(P<0.01、P<0.05和P<0.01),而治疗组、预防治疗组与正常对照组没有显著差异。
2.3血浆GLU含量(pg/ml):注射ALX后第93天,糖尿病对照组、治疗组血浆GLU显著或非常显著高于正常对照组(P<0.01、P<0.05),预防治疗组与正常对照组之间无显著差异(p>0.05),但预防治疗组显著低于糖尿病对照组(P<0.05)。
3.大鼠血清SOD、MDA含量测定结果:注射ALX后第33天和93天,糖尿病对照组、治疗组、预防治疗组SOD含量非常显著低于正常对照组(P<0.01),MDA含量非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组SOD含量非常显著高于糖尿病对照组(p<0.01),MDA含量显著或非常显著低于糖尿病对照组(P<0.05或p<0.01)。
4.大鼠血清TG、TC、LDL测定结果:注射ALLOX后第93天,治疗组、预防治疗组血清TG、TC、LDL含量显著或非常显著低于糖尿病对照组(P<0.05或P<0.01)。
5.大鼠血清Cr和BUN测定结果:注射ALLOX后第93天,糖尿病对照组、治疗组、预防治疗组Cr和BUN含量非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组显著或非常显著低于糖尿病对照组(p<0.05或p<0.01)。
6.大鼠胰岛病理结构变化:光镜下观察,正常对照组大鼠胰岛呈着色淡浅的细胞团,边界清楚,形态规则,胰岛内细胞排列整齐,胰岛数及岛内细胞数较多,大小一致分布均匀,细胞核呈紫蓝色,大而圆,核清晰,核仁明显,胞浆丰富呈淡粉色;糖尿病对照组大鼠胰岛细胞结构紊乱,胰岛数及岛内细胞数减少,细胞形态不规则,多数胞核变形,固缩,大小不等,出现空泡变性;治疗组胰岛数及岛内细胞数较糖尿病对照组为多,形态较为规则,少部分核固缩;预防治疗组大鼠胰岛细胞形态接近正常组,边界较清楚,细胞排列较整齐,胰岛数及岛内细胞数多于治疗组,细胞分布均匀,核大小基本相等,无核固缩。
7.大鼠胰岛组织细胞胞浆胰岛素(INS)和胰高血糖素(GLU)免疫组化结果:胞浆镜下观察,正常对照组胰岛B细胞分泌INS最多,遍布胰岛中央,分泌颗粒丰富,胞浆着色深黄;糖尿病对照组胰岛B细胞分泌INS颗粒最少,胞浆着色较浅;治疗组胰岛B细胞分泌INS颗粒数量较糖尿病对照组微多,胞浆着色浅黄;预防治疗组B细胞分泌INS颗粒数量接近正常对照组,胞浆着色较深。正常对照组胰岛A细胞分泌GLU颗粒数量少均匀分布胰岛周围,着色较浅;糖尿病对照组胰岛A细胞分泌GLU颗粒丰富,大部分已经进入到胰岛中间区,胞浆着色深黄;治疗组A细胞分泌GLU颗粒数量较糖尿病对照组少,一部分进入到胰岛中间区,胞浆着色淡黄;预防治疗组A细胞分泌GLU颗粒大部分分布在胰岛周围,分泌颗粒较多,胞浆着色浅黄。
8.大鼠胰腺组织胰岛B细胞凋亡:正常对照组可见少数凋亡细胞;糖尿病对照组有较多凋亡细胞;治疗组和预防治疗组也可见一些凋亡细胞,但明显少于糖尿病对照组;预防治疗组B细胞凋亡数也较治疗组少。
结论:
1.大鼠空腹血糖、糖耐量、糖化血红蛋白测定结果表明,该糖尿病复合营养酸奶具有明显改善糖代谢和降血糖效应;
2.胰岛病理结构观察、胰岛组织细胞胞浆免疫组化和胰岛B细胞凋亡实验结果表明,糖尿病复合营养酸奶对ALX所致糖尿病大鼠胰岛结构和功能的损伤具有保护作用;
3.血清MDA和SOD含量的测定表明,糖尿病复合营养酸奶能明显减轻糖尿病大鼠机体过氧化损伤,此作用可能是糖尿病复合营养酸奶对胰岛结构和功能具有保护作用的机制之一;
4.血清TG、TC、LDL测定结果表明,糖尿病复合营养酸奶具有改善糖尿病大鼠脂代谢紊乱,此作用对防治糖尿病血清疾病并发症是有益的作用;
5.研究结果还表明,该糖尿病复合营养酸奶具有减轻ALX所致糖尿病大鼠肾脏损伤、预防感染和降低死亡率的作用;
6.由于预防治疗组动物在注射ALX建立糖尿病模型前即以摄入糖尿病复合营养酸奶,动物实验的各项研究指标显示预防治疗组较治疗组具有更明显的效果,表明该糖尿病复合营养酸奶对预防糖尿病具有潜在的应用前景;
7.该糖尿病复合营养酸奶对糖尿病大鼠具有的上述作用可能是配方组成中含有的多种营养功效成分综合作用的结果;
OBJECTIVE:
Diabetes mellitus(DM) was a endocrine metabolic disease and severe harm the health of body. The attack rate and death rate of DM increased more and more quickly. At present there were 1.5 hundred million diabetic in the world .It was more than 40000 thousand diabetic in china. Because of the decrease of islet cell and action of islet cell ,it caused the disorder of fat ,protein, water and electrolyte and produced severe complication ,for example, coronary heart disease、cerebrovascular disease、peripheral vascular disease、retinopathy and renal vascular disease ect. More than fifty percent of diabetic who succumbed to coronary heart disease. Therefore, it was very important significance for us to improve the health status of diabetic and enhence the quality of life of diabetic and prevent the complication of diabetic and promote the rehabilitation of diabetic. So the ideal method of searching for treating and controlling diabetic was very important topic of medical science boundary. So in this study we observed and researched the protective effects of Compound acidophilous milk for diabetic on Alloxan induced pancreatic gland damage and glycometabolism in rats and cut down the influence of pancreatic tissue histomorphology and lipid peroxidation and pancreatic island B apoptotic .We also Attempted to provid some significant experimental based for developing and exploiting efficiency food.
METHODS:
120 Adult Wistar rats(190±10) were randomly divided into Control group ,DM group ,therapy group and protective therapy group ,30 in each group. Each group were fed a basic diet and Control group were intragastric administration with Sodium Chloride, DM group and therapy group were intragastric administration with ordinary acidophilous milk, protective therapy group were intragastric administration with compound acidophilous milk of diabetic .Each guoup was intragastric administration every other day. After 15 days, the rats of DM group, therapy group and protective therapy group have been given a single shot of alloxan (50mg per 1 kg), while Control group have been given a single shot of Sodium Chloride (50mg per 1 kg) . After 5 days , determining fasting 4 hours blood sugar,then the rats of blood sugar more than 13.8 mmol/L were diabetic model rats. After the diabetic model have been induced by alloxan , the rats of therapy group were intragastric administration with compound acidophilous milk of diabetic ,and other groups have not been changed . In 30 th days and 90th days after every group were given a single shot of alloxa determines fasting 4 hours blood sugar and In 6th days、32 th days and 92th days after every group were given a single shot of alloxa carbohydrate tolerance test .In 33th days and 93th days after every group were given a single shot of alloxa, randomly drawing out 10 rats in every groups that were killed and the pancreatic gland samples and blood serum were collected .Tentative index include INS ,C-P ,MDA ,SOD ,Cr ,BUN ,TG ,TC , LDL in blood serum and GLU, glycosylated hemoglobin in blood plasm and so on.The pancreatic gland morphological structure was also obserced under lingt microscope .The degree of apoptosis was assessed with the terminal deoxynucleotidy1 transferase (TdT)-mediated d UTP nick end labeling (TUNEL) by using an in situ Apoptosis Detection Kit.The expression of INS in the cytoplasm of B cell and GLU in the cytoplasm of A cell was detected by immunohistochemistry.
RESULTS:
1.Effect of compound acidophilous milk for diabetic on blood-fasting sugar and sugar tolerance and glycosylated hemoglobin
1.1 evel of FBS(mmol/L):
1.1.1 th days after every group were given alloxa, the level of FBS in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of FBS in the protective therapy group is lower than the DM group,the therapy group(P<0.01);
1.1.2 0th days and 90th days after every group were given alloxa, the level of FBS in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of FBS in the therapy group and the protective therapy group is lower than the DM group (P<0.01);
1.2 el of sugar tolerance(mmol/L): on 6th day、32th day、92th day , compare with Control group, the difference of sugar tolerancein 0.5h、1.0h、1.5h、2.0h of DM group , therapy group, protective therapy group is very significantly (P<0.05) while compare with DM group ,the difference of sugar tolerance in 0.5h、1.0h、1.5h、2.0h of therapy group, protective therapy group is very significantly (P<0.01). sugar tolerance of 0.5h、1.0h、1.5h、2.0h of 6th and sugar tolerance of 1.0h、1.5h of 32th of protective therapy group were different from DM group’and therapy group’,and the difference were very significantly(P<0.05)(p<0.01).
1.3 evel of glycosylated hemoglobin(%):93th days after every group were given alloxa, the level of glycosylated hemoglobin in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of glycosylated hemoglobin in the therapy group and the protective therapy group is lower than the DM group (P<0.01);
2.Effect of compound acidophilous milk for diabetic on pancreatic function
2.1 evel of serum INS (μIU/ml):33 th days and 93th days after every group were given alloxa, the level of INS in the DM group ,the therapy group and the protective therapy group is more lower than control group(P<0.01), but the level of INS in the therapy group and the protective therapy group is very significantly higher than the DM group(P<0.05 and P<0.01);
2.2 evel of serum C-P(ng/ml): 33th days and 93th days after every group were given alloxa, the level of C-P in the control group, the therapy group and the protective therapy group is very significantly higher than the DM group(P<0.01、P<0.05 and P<0.01);furthermore, the level of C-P in the therapy group and the protective therapy group doses not have significant difference.
2.3 evel of plasm GLU(pg/ml): 93th days after every group were given alloxa , the Level of plasm GLU in the control group and the protective therapy group is very significantly lower than the DM group(P<0.01 and P<0.05);the Level of plasm GLU in the therapy group is higher control group ,but also significantly lower than the DM group(P<0.05).
3.Effect of compound acidophilous milk for diabetic on serum SOD、MDA 33th days and 93 days after every group were given alloxa,the level of SOD in DM group, the therapy group and the protective therapy group is very significantly lower than control group(P<0.01),the level of MDA in the DM group,the therapy group and the protective therapy group is more higher than control group(P<0.01),but the level of SOD in the therapy group and the protective therap is higher than DM group(P<0.01), the level of MDA in the therapy group and the protective therapy group is very significantly lower than the DM group (p<0.05 or P<0.01);
4.Effect of compound acidophilous milk for diabetic on sersum TG、TC and LDL 93 days after every group were given alloxa,the level of TG、TC and LDL in the therapy group and the protective therapy group is very significantly lower than DM group(P<0.05 and P<0.01).
5.Effect of compound acidophilous milk for diabetic on sersum Cr and BUN(μmol/L) 93 days after every group were given alloxa, the level of serum Cr and BUN in the DM group ,the therapy group , the protective therapy group is very significantly higher than control group(P<0.01),but the level of serum Cr and BUN in the therapy group , the protective therapy group is very significantly lower than DM group(P<0.05,P<0.01)
6.Effent of compound acidophilous milk of diabetic on the pancreatic gland morpholological structure
Examination of histological structure under a light microscope indicated that pancreatic islets of control group were colored to pale colour and bouncary distinctness, morphous regulation, lineing up in order.The number of pancreatic islet and isle cells in control group is more than DM group .Cell nucleus of control group were blue and limpid. Chromatospherite is conspicuous. Pancreatic islet cellular structure of DM group disordered and morphous is inordinate. Lots of cell nucleus of DM group were deformed, pycnotic and emerged to vacuolar degeneration. The number of pancreatic islet and isle cells in the therapy group was more than DM group but lower than control group. A little of cell nucleus were deformed and pycnotic. Islet cells of the protective therapy group were closed to control group and bouncary distinctness, morphous regulation, lineing up in order.
7.Express of the INS and GLU in the endochylema of pancreatic Islet
Examination of immunohistochemistry under a light microscope indicated that Islet B cells of control group secretory INS were more than other groups and secreted lots of grains .Endochylema of cells were colored to deep color. Islet B cells of DM group secreted a little of grains. Endochylema of cells were colored to tint. The number and color of grains in the therapy group was more than DM group , furthermore ,the number and color of grains in the protective therapy group was closed to control group.
Islet A cells of control group secretory GLU were less than other groups and the GLU grains situated In the fringe of pancreatic islet .Endochylema of cells were colored to tint color. Islet A cells of DM group secreted lots of grains that a part of have intruded into the middle of Islet. The number and color of grains in the therapy group was less than DM group , furthermore ,the number and color of grains in the protective therapy group was closed to control group.
8.The alloxan-induced TUNEL-positive B cells in pancreatic islet in the DM group were particularly more than in the therapy group and protective therapy group. Further,there was only a few TUNEL-positive B cells in the control group.
CONCLUSIONS:
1.The result of FBS, OTGC ,HGb of the rats indicated compound acidophilous milk of diabetic could obviously improve glycometabolism and decease effect of blood sugar ;
2.The observation of pancreatic island patho-structure, immunity class of pancreatic island histiocyte endochylema and the B apoptosis of pancreatic island indicated compound acidophilous milk of diabetic have protection for the structure and function of pancreatic island;
3.The result of sersum MDA and SOD indicated that compound acidophilous milk of diabetic could bviously lessen oxidative damage of organism in DM rats. It may be that compound acidophilous milk of diabetic had protection for the structure and function of pancreatic island;
4.The result of serumtTG, TC and LDL indicated that compound acidophilous milk of diabetic could have action of improving disorder of lipid metabolism in DM rats;
5.The finding indicated compound acidophilous milk of diabetic could have the protection of lessening kidney damage of the DM, infection prevention and cutting down death rate rats;
6.Beause of protective therapy group were intragastric administration with compound acidophilous milk of diabetic before founding DM model ,every index of animal experiment indicated protective therapy group had best effect .This indicated that compound acidophilous milk of diabetic had potential application prospect for preventing diabetes;
7.Compound acidophilous milk of diabetic had above-mentioned effect that because of there were lots of nutrition efftective ingredient in prescription of compound acidophilous milk of diabetic;
糖尿病(DM)是一种全球范围内严重影响人类健康的常见内分泌代谢性疾病,其发病率和死亡率近年来呈逐步上升趋势。目前全世界已有糖尿病人近1.5亿,我国实际患病人数已达4000万人以上。DM主要是由于胰岛B细胞减少或B细胞功能减退引起的糖、脂肪、蛋白质、水及电解质代谢紊乱,并可由此产生各种并发症,如由大血管或微血管病变所引起的冠心病、脑血管疾病、视网膜病变、肾血管病变等,其中糖尿病患者有50%以上死于冠心病。因此,有效的改善糖尿病的代谢紊乱对预防糖尿病并发症、促进疾病康复和提高病人生活质量都具有十分重要的意义,而努力探索和积极寻找治疗和控制糖尿病的理想方法是医学界关注的重要课题。本研究以生物发酵作为切入点,通过观察我们所研制的糖尿病复合营养酸奶对保护糖尿病大鼠的胰腺功能、改善糖代谢、降低血糖血脂及对胰腺组织形态学、脂质过氧化状态和胰岛B细胞凋亡的影响,为糖尿病患者功效食品的研制提供实验依据。
方法:
清洁级成年Wister大鼠(雌雄各半)120只,体重190±10克,随机分为正常对照组、糖尿病对照组、治疗组、预防治疗组四组,每组30只。在常规饲料自由进食基础上,正常对照组给予生理盐水灌喂,糖尿病对照组和治疗组给予普通酸奶灌喂,预防治疗组给予糖尿病复合营养酸奶灌喂,隔日一次,每次2毫升,喂养至第15天按四氧嘧啶(ALX)50mg/k.w尾静脉注射制作糖尿病模型,正常对照组注射生理盐水。5天后测其空腹4小时后血糖,将血糖值>13.8mmol/L的大鼠做为糖尿病模型大鼠。糖尿病模型制作成功后除治疗组动物灌喂液由原来普通酸奶改为糖尿病复合营养酸奶外,其他组动物喂养方式不变,继续分组喂养,并于注射ALX后第30天、第90天再次进行空腹4小时血糖测定,注射ALX后第6天、第32天、第92天进行糖耐量测定,注射ALX后第33天、第93天分别从各组大鼠中随机取10只大鼠进行腹主动脉取血、处死大鼠取材进行以下指标测定:血清中的胰岛素(INS)、C-肽(C-P)、丙二醛(MDA)、超氧化物歧化酶(SOD)、肌酐(Cr)、尿素氮(BUN)、甘油三酯(TG)、总胆固醇(TC)和低密度脂蛋白(LDL)的含量,血浆中胰高血糖素(GLU)和糖化血红蛋白(GHb)含量,光镜下观察病理结构改变,TUNEL法测定胰岛B细胞凋亡,免疫组化法观察INS和GLU分泌的表达。
结果:
1.大鼠空腹血糖、糖耐量及糖化血红蛋白测定结果:
1.1 FBS水平(mmol/L):
1.1.1注射ALX后第5天,糖尿病对照组、治疗组、预防治疗组FBS非常显著高于正常对照组(P<0.01),预防治疗组非常显著低于糖尿病对照组和治疗组(P<0.01)。
1.1.2注射ALX后第30天和90天,糖尿病对照组、治疗组、预防治疗组非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组非常显著低于糖尿病对照组(P<0.01)。
1.2糖耐量测定结果(mmol/L):注射ALX后第6天、第32天、第92天糖尿病对照组、治疗组、预防治疗组动物0.5h、1.0h、1.5h、2.0h血糖测定值显著或非常显著高于正常对照组(P<0.05或p<0.01),但治疗组、预防治疗组第32天、92天上述各时相点血糖测定值非常显著低于糖尿病对照组(P<0.01),并且预防治疗组第6天上述各时相点血糖测定值非常显著低于治疗组(P<0.01)。
1.3糖化血红蛋白测定结果(%):注射ALX后第93天,糖尿病对照组、治疗组、预防治疗组糖化血红蛋白含量非常显著高于正常对照组(P<0.01),但治疗组和预防治疗组显著低于糖尿病对照组(P<0.01)。
2.大鼠血清INS、C-P、GLU测定结果:
2.1血清INS含量(μIU/ml):注射ALX后第33天、第93天,糖尿病对照组、治疗组、预防治疗组INS含量非常显著低于正常对照组(P<0.01),但治疗组、预防治疗组显著或非常显著高于糖尿病对照组(P<0.05或p<0.01)。
2.2血清C-P含量(ng/ml):注射ALX后第33天和93天,正常组、治疗组、预防治疗组C-P含量显著或非常显著高于糖尿病对照组(P<0.01、P<0.05和P<0.01),而治疗组、预防治疗组与正常对照组没有显著差异。
2.3血浆GLU含量(pg/ml):注射ALX后第93天,糖尿病对照组、治疗组血浆GLU显著或非常显著高于正常对照组(P<0.01、P<0.05),预防治疗组与正常对照组之间无显著差异(p>0.05),但预防治疗组显著低于糖尿病对照组(P<0.05)。
3.大鼠血清SOD、MDA含量测定结果:注射ALX后第33天和93天,糖尿病对照组、治疗组、预防治疗组SOD含量非常显著低于正常对照组(P<0.01),MDA含量非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组SOD含量非常显著高于糖尿病对照组(p<0.01),MDA含量显著或非常显著低于糖尿病对照组(P<0.05或p<0.01)。
4.大鼠血清TG、TC、LDL测定结果:注射ALLOX后第93天,治疗组、预防治疗组血清TG、TC、LDL含量显著或非常显著低于糖尿病对照组(P<0.05或P<0.01)。
5.大鼠血清Cr和BUN测定结果:注射ALLOX后第93天,糖尿病对照组、治疗组、预防治疗组Cr和BUN含量非常显著高于正常对照组(P<0.01),但治疗组、预防治疗组显著或非常显著低于糖尿病对照组(p<0.05或p<0.01)。
6.大鼠胰岛病理结构变化:光镜下观察,正常对照组大鼠胰岛呈着色淡浅的细胞团,边界清楚,形态规则,胰岛内细胞排列整齐,胰岛数及岛内细胞数较多,大小一致分布均匀,细胞核呈紫蓝色,大而圆,核清晰,核仁明显,胞浆丰富呈淡粉色;糖尿病对照组大鼠胰岛细胞结构紊乱,胰岛数及岛内细胞数减少,细胞形态不规则,多数胞核变形,固缩,大小不等,出现空泡变性;治疗组胰岛数及岛内细胞数较糖尿病对照组为多,形态较为规则,少部分核固缩;预防治疗组大鼠胰岛细胞形态接近正常组,边界较清楚,细胞排列较整齐,胰岛数及岛内细胞数多于治疗组,细胞分布均匀,核大小基本相等,无核固缩。
7.大鼠胰岛组织细胞胞浆胰岛素(INS)和胰高血糖素(GLU)免疫组化结果:胞浆镜下观察,正常对照组胰岛B细胞分泌INS最多,遍布胰岛中央,分泌颗粒丰富,胞浆着色深黄;糖尿病对照组胰岛B细胞分泌INS颗粒最少,胞浆着色较浅;治疗组胰岛B细胞分泌INS颗粒数量较糖尿病对照组微多,胞浆着色浅黄;预防治疗组B细胞分泌INS颗粒数量接近正常对照组,胞浆着色较深。正常对照组胰岛A细胞分泌GLU颗粒数量少均匀分布胰岛周围,着色较浅;糖尿病对照组胰岛A细胞分泌GLU颗粒丰富,大部分已经进入到胰岛中间区,胞浆着色深黄;治疗组A细胞分泌GLU颗粒数量较糖尿病对照组少,一部分进入到胰岛中间区,胞浆着色淡黄;预防治疗组A细胞分泌GLU颗粒大部分分布在胰岛周围,分泌颗粒较多,胞浆着色浅黄。
8.大鼠胰腺组织胰岛B细胞凋亡:正常对照组可见少数凋亡细胞;糖尿病对照组有较多凋亡细胞;治疗组和预防治疗组也可见一些凋亡细胞,但明显少于糖尿病对照组;预防治疗组B细胞凋亡数也较治疗组少。
结论:
1.大鼠空腹血糖、糖耐量、糖化血红蛋白测定结果表明,该糖尿病复合营养酸奶具有明显改善糖代谢和降血糖效应;
2.胰岛病理结构观察、胰岛组织细胞胞浆免疫组化和胰岛B细胞凋亡实验结果表明,糖尿病复合营养酸奶对ALX所致糖尿病大鼠胰岛结构和功能的损伤具有保护作用;
3.血清MDA和SOD含量的测定表明,糖尿病复合营养酸奶能明显减轻糖尿病大鼠机体过氧化损伤,此作用可能是糖尿病复合营养酸奶对胰岛结构和功能具有保护作用的机制之一;
4.血清TG、TC、LDL测定结果表明,糖尿病复合营养酸奶具有改善糖尿病大鼠脂代谢紊乱,此作用对防治糖尿病血清疾病并发症是有益的作用;
5.研究结果还表明,该糖尿病复合营养酸奶具有减轻ALX所致糖尿病大鼠肾脏损伤、预防感染和降低死亡率的作用;
6.由于预防治疗组动物在注射ALX建立糖尿病模型前即以摄入糖尿病复合营养酸奶,动物实验的各项研究指标显示预防治疗组较治疗组具有更明显的效果,表明该糖尿病复合营养酸奶对预防糖尿病具有潜在的应用前景;
7.该糖尿病复合营养酸奶对糖尿病大鼠具有的上述作用可能是配方组成中含有的多种营养功效成分综合作用的结果;
OBJECTIVE:
Diabetes mellitus(DM) was a endocrine metabolic disease and severe harm the health of body. The attack rate and death rate of DM increased more and more quickly. At present there were 1.5 hundred million diabetic in the world .It was more than 40000 thousand diabetic in china. Because of the decrease of islet cell and action of islet cell ,it caused the disorder of fat ,protein, water and electrolyte and produced severe complication ,for example, coronary heart disease、cerebrovascular disease、peripheral vascular disease、retinopathy and renal vascular disease ect. More than fifty percent of diabetic who succumbed to coronary heart disease. Therefore, it was very important significance for us to improve the health status of diabetic and enhence the quality of life of diabetic and prevent the complication of diabetic and promote the rehabilitation of diabetic. So the ideal method of searching for treating and controlling diabetic was very important topic of medical science boundary. So in this study we observed and researched the protective effects of Compound acidophilous milk for diabetic on Alloxan induced pancreatic gland damage and glycometabolism in rats and cut down the influence of pancreatic tissue histomorphology and lipid peroxidation and pancreatic island B apoptotic .We also Attempted to provid some significant experimental based for developing and exploiting efficiency food.
METHODS:
120 Adult Wistar rats(190±10) were randomly divided into Control group ,DM group ,therapy group and protective therapy group ,30 in each group. Each group were fed a basic diet and Control group were intragastric administration with Sodium Chloride, DM group and therapy group were intragastric administration with ordinary acidophilous milk, protective therapy group were intragastric administration with compound acidophilous milk of diabetic .Each guoup was intragastric administration every other day. After 15 days, the rats of DM group, therapy group and protective therapy group have been given a single shot of alloxan (50mg per 1 kg), while Control group have been given a single shot of Sodium Chloride (50mg per 1 kg) . After 5 days , determining fasting 4 hours blood sugar,then the rats of blood sugar more than 13.8 mmol/L were diabetic model rats. After the diabetic model have been induced by alloxan , the rats of therapy group were intragastric administration with compound acidophilous milk of diabetic ,and other groups have not been changed . In 30 th days and 90th days after every group were given a single shot of alloxa determines fasting 4 hours blood sugar and In 6th days、32 th days and 92th days after every group were given a single shot of alloxa carbohydrate tolerance test .In 33th days and 93th days after every group were given a single shot of alloxa, randomly drawing out 10 rats in every groups that were killed and the pancreatic gland samples and blood serum were collected .Tentative index include INS ,C-P ,MDA ,SOD ,Cr ,BUN ,TG ,TC , LDL in blood serum and GLU, glycosylated hemoglobin in blood plasm and so on.The pancreatic gland morphological structure was also obserced under lingt microscope .The degree of apoptosis was assessed with the terminal deoxynucleotidy1 transferase (TdT)-mediated d UTP nick end labeling (TUNEL) by using an in situ Apoptosis Detection Kit.The expression of INS in the cytoplasm of B cell and GLU in the cytoplasm of A cell was detected by immunohistochemistry.
RESULTS:
1.Effect of compound acidophilous milk for diabetic on blood-fasting sugar and sugar tolerance and glycosylated hemoglobin
1.1 evel of FBS(mmol/L):
1.1.1 th days after every group were given alloxa, the level of FBS in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of FBS in the protective therapy group is lower than the DM group,the therapy group(P<0.01);
1.1.2 0th days and 90th days after every group were given alloxa, the level of FBS in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of FBS in the therapy group and the protective therapy group is lower than the DM group (P<0.01);
1.2 el of sugar tolerance(mmol/L): on 6th day、32th day、92th day , compare with Control group, the difference of sugar tolerancein 0.5h、1.0h、1.5h、2.0h of DM group , therapy group, protective therapy group is very significantly (P<0.05) while compare with DM group ,the difference of sugar tolerance in 0.5h、1.0h、1.5h、2.0h of therapy group, protective therapy group is very significantly (P<0.01). sugar tolerance of 0.5h、1.0h、1.5h、2.0h of 6th and sugar tolerance of 1.0h、1.5h of 32th of protective therapy group were different from DM group’and therapy group’,and the difference were very significantly(P<0.05)(p<0.01).
1.3 evel of glycosylated hemoglobin(%):93th days after every group were given alloxa, the level of glycosylated hemoglobin in the DM group,the therapy group and the protective therapy group increased very significantly more than control group(P<0.01), but the level of glycosylated hemoglobin in the therapy group and the protective therapy group is lower than the DM group (P<0.01);
2.Effect of compound acidophilous milk for diabetic on pancreatic function
2.1 evel of serum INS (μIU/ml):33 th days and 93th days after every group were given alloxa, the level of INS in the DM group ,the therapy group and the protective therapy group is more lower than control group(P<0.01), but the level of INS in the therapy group and the protective therapy group is very significantly higher than the DM group(P<0.05 and P<0.01);
2.2 evel of serum C-P(ng/ml): 33th days and 93th days after every group were given alloxa, the level of C-P in the control group, the therapy group and the protective therapy group is very significantly higher than the DM group(P<0.01、P<0.05 and P<0.01);furthermore, the level of C-P in the therapy group and the protective therapy group doses not have significant difference.
2.3 evel of plasm GLU(pg/ml): 93th days after every group were given alloxa , the Level of plasm GLU in the control group and the protective therapy group is very significantly lower than the DM group(P<0.01 and P<0.05);the Level of plasm GLU in the therapy group is higher control group ,but also significantly lower than the DM group(P<0.05).
3.Effect of compound acidophilous milk for diabetic on serum SOD、MDA 33th days and 93 days after every group were given alloxa,the level of SOD in DM group, the therapy group and the protective therapy group is very significantly lower than control group(P<0.01),the level of MDA in the DM group,the therapy group and the protective therapy group is more higher than control group(P<0.01),but the level of SOD in the therapy group and the protective therap is higher than DM group(P<0.01), the level of MDA in the therapy group and the protective therapy group is very significantly lower than the DM group (p<0.05 or P<0.01);
4.Effect of compound acidophilous milk for diabetic on sersum TG、TC and LDL 93 days after every group were given alloxa,the level of TG、TC and LDL in the therapy group and the protective therapy group is very significantly lower than DM group(P<0.05 and P<0.01).
5.Effect of compound acidophilous milk for diabetic on sersum Cr and BUN(μmol/L) 93 days after every group were given alloxa, the level of serum Cr and BUN in the DM group ,the therapy group , the protective therapy group is very significantly higher than control group(P<0.01),but the level of serum Cr and BUN in the therapy group , the protective therapy group is very significantly lower than DM group(P<0.05,P<0.01)
6.Effent of compound acidophilous milk of diabetic on the pancreatic gland morpholological structure
Examination of histological structure under a light microscope indicated that pancreatic islets of control group were colored to pale colour and bouncary distinctness, morphous regulation, lineing up in order.The number of pancreatic islet and isle cells in control group is more than DM group .Cell nucleus of control group were blue and limpid. Chromatospherite is conspicuous. Pancreatic islet cellular structure of DM group disordered and morphous is inordinate. Lots of cell nucleus of DM group were deformed, pycnotic and emerged to vacuolar degeneration. The number of pancreatic islet and isle cells in the therapy group was more than DM group but lower than control group. A little of cell nucleus were deformed and pycnotic. Islet cells of the protective therapy group were closed to control group and bouncary distinctness, morphous regulation, lineing up in order.
7.Express of the INS and GLU in the endochylema of pancreatic Islet
Examination of immunohistochemistry under a light microscope indicated that Islet B cells of control group secretory INS were more than other groups and secreted lots of grains .Endochylema of cells were colored to deep color. Islet B cells of DM group secreted a little of grains. Endochylema of cells were colored to tint. The number and color of grains in the therapy group was more than DM group , furthermore ,the number and color of grains in the protective therapy group was closed to control group.
Islet A cells of control group secretory GLU were less than other groups and the GLU grains situated In the fringe of pancreatic islet .Endochylema of cells were colored to tint color. Islet A cells of DM group secreted lots of grains that a part of have intruded into the middle of Islet. The number and color of grains in the therapy group was less than DM group , furthermore ,the number and color of grains in the protective therapy group was closed to control group.
8.The alloxan-induced TUNEL-positive B cells in pancreatic islet in the DM group were particularly more than in the therapy group and protective therapy group. Further,there was only a few TUNEL-positive B cells in the control group.
CONCLUSIONS:
1.The result of FBS, OTGC ,HGb of the rats indicated compound acidophilous milk of diabetic could obviously improve glycometabolism and decease effect of blood sugar ;
2.The observation of pancreatic island patho-structure, immunity class of pancreatic island histiocyte endochylema and the B apoptosis of pancreatic island indicated compound acidophilous milk of diabetic have protection for the structure and function of pancreatic island;
3.The result of sersum MDA and SOD indicated that compound acidophilous milk of diabetic could bviously lessen oxidative damage of organism in DM rats. It may be that compound acidophilous milk of diabetic had protection for the structure and function of pancreatic island;
4.The result of serumtTG, TC and LDL indicated that compound acidophilous milk of diabetic could have action of improving disorder of lipid metabolism in DM rats;
5.The finding indicated compound acidophilous milk of diabetic could have the protection of lessening kidney damage of the DM, infection prevention and cutting down death rate rats;
6.Beause of protective therapy group were intragastric administration with compound acidophilous milk of diabetic before founding DM model ,every index of animal experiment indicated protective therapy group had best effect .This indicated that compound acidophilous milk of diabetic had potential application prospect for preventing diabetes;
7.Compound acidophilous milk of diabetic had above-mentioned effect that because of there were lots of nutrition efftective ingredient in prescription of compound acidophilous milk of diabetic;
引文
1. Battell ML, Yuen VG, Verma S and McNeill JH. Other models of type I diabetes. In: H. John McNeill, Editor, Experimental Models of Diabetes, CRC, USA ,1999:219–229.
2. Tiedge M, Richter T and Lenzen S. Importance of cysteine residues for the stability and catalytic activity of human pancreatic beta cell glucokinase[J].Arch Biochem Biophys, 2000, 375:251–260.
3. Bloch KO, Zemel R, Bloch OV, Grief H and Vardi P. Streptozotocin and alloxan-based selection improves toxin resistance of insulin-producing RINm cells[J]. Int J Exp Diabetes Res, 2000, 1 : 211–219.
4. Gai W, Schott-Ohly P, Gleichmann H, et al. Differential target molecules for toxicity induced by streptozotocin and alloxan in pancreatic islets of mice in vitro[J]. Exp Clin Endocrinol Diabetes ,2004, 112 : 29–37
5. Okamoto H. New aspects to the functioning and regeneration of pancreatic β-cells[J]. Adv Exp Med. Biol ,1997, 426: 301–311.
6. Sentman ML, Jonsson LM and Marklund SL. Enhanced alloxan-induced beta-cell damage and delayed recovery from hyperglycemia in mice lacking extracellular - superoxide dismutase[J]. Free Radic Biol Med, 1999, 27:790–796.
7. 张晓波,姚海涛,杨子安,等.有机铬对实验性糖尿病大鼠胰腺保护作用的观察 [J]. 黑龙江医药科学,2004 ,27(5):1-2
8. 胡晓斐,张永奎,孙文均,等 .鱼提取物(鱼酱油)对大鼠血糖及体重影响的实验研究[J].中华航海医学与高气压医学杂志,2006,13(1):23-25
9. DeFronzo RA. Diabetes ,1988, 37:667–687.
10. Weir GC, Laybutt DR, Kaneto etc. Diabetes, 2001 ,50 (Suppl.1): 154–159.
11. Rossetti L, Giaccari A and De Fronzo RA. Diabetes Care ,1990, 13: 610–630.
12. Donath MY, Gross DJ, Cerasi E and Kaiser N. Diabetes ,1999, 48: 738–744.
13. McGarry JD and Dobbins RL .Diabetologia, 1999, 42: 128–138.
14. Lorenzo A, Razzaboni B, Weir GC and Yankner BA. Nature, 1994, 368:756–760.
15. Polonsky KS, Sturis J and Bell GI. Engl N .Med, 1996, 334: 777–783.
16. Brownlee M. Nature ,2001 ,414: 813–820.
17. Robertson RP. Biol Chem, 2004, 279:42351–42354.
18. Thornalley P, Wolff S, Crabbe J and Stern A. Biochim Biophys Acta ,1984, 797:276–287.
19. 南征、高彦彬、钱秋海主编. 糖尿病中西医综合治疗, 北京:人民卫生出版社, 2002: 453
20. Clark A , Wells CA , Buley ID , et al . Islet amyloid , increased A - cells ,reduced B - cells and exocrine fibrosis : quantitative changes in the pancreas in type 2 diabetes. Diabetes Res , 1988 ,9 :151
21. Butler AE , Janson J , Ritzel R ,et al . Accelerated apoptosis overcomes in creased replication to causeβ- cell loss in diabetes in mice transgenic for h - IAPP. Diabetes , 2002 ,51 (Suppl 2) :A7
22. Butler AE , Janson J , Walter C , et al . Increased B - cell apoptosis pre2vents adaptive increase in B - cell mass in mouse model of type 2 dia2betes : evidence for role of islet amyloid formation rather than direct action of amyloid. Diabetes , 2003 , 52 :2304
23. Butler AE , Janson J , Bonner - Weir S ,et al . Beta - cell deficit and in 2 creased beta - cell apoptosis in humans with type 2 diabetes. Diabetes ,2003 , 52 :102
24. Biarnes M, Montolio M, Nacher V , et al .β- cell death and mass in syn geneically transplanted islets exposed to short - and long - term hyper2 glycemia. Diabetes , 2002 , 51 :66
25. Federici M, Hribal M, Perego L , et al . High glucose causes apoptosis in cultured human pancreatic islets of langerhans. Diabetes , 2001 , 50 :1290
26. Kaneto H, Fujii J and Myint T etal. Biochem, 1996, 320 (Pt. 3):855–863.
27. 张娅婕, 陈秋丽, 甘振威, 等 1 Cr3 + 对糖尿病小鼠降血糖作用的研究[ J ]吉林大学学报: 医学版, 2004 , 30 ( 3) :379-380
28. 吴蕴棠,车素萍,孙忠,等. 锌、硒、铬对糖尿病大鼠糖、脂代谢调节作用[J ] . 中国公共卫生,2004 ,20 (9) :1083 - 1085.
29. Monnier VM. Gerontol ,1990, 45: B105–B111.
30. Tremblay F , Dubois MJ , Marette A. Regulationof GLUT4 traffic and function by insulin and contraction in skeletal muscle[J] . Front Biosci,2003,8:1072―1084.
31. Evans JL, Goldfine ID, Maddux BA and Grodsky GM. Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes[J]. Endocr Rev ,2002, 23: 599–622.
32. Zelko IN, Mariani TJ and Folz JR. Superoxide dismutase multigene family: a comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression[J]. Free Radic Biol Med, 2002, 33:337–349.
33. Fattman CL, Schaefer LM and Oury TD. Extracellular superoxide dismutase in biology and medicine[J]. Free Radic Biol Med, 2003, 35:236–256.
34. Adachi T, Inoue M, Hara H, Maehata E and Suzuki S. Relationship of plasma extracellular-superoxide dismutase level with insulin resistance in type 2 diabetic patients[J]. Endocrinol, 2004, 181: 413–417
35. Kurien BT, Scofield RH. Free radical mediated peroxidative damage in systemic lupus erythematosu[J].Life Sci,2003,73(13):1655-1666.
36. Malarkodi KP ,Balachandar AV, Sivaprasad R,et al. Prophylactic effect of lipoic acid against adriamycin-induced peroxidative damages in rat kidney[J]. Ren Fail, 2003, 25(3):367-377.
37. Hail ED, Andrus PK, Oostveen JA, et al. Relationship of oxygen radical-induced lipid peroxidative damage to disease onset and progression in a transgenic model of familial ALS[J]. JNeurosci Res, 1998, 53(1):66-77.
38. Isono M, Mogyorosi A ,Han DC , et al . Stimulation of TGF2beta type Ⅱreceptor by high glucose in mouse mesangial cells and in diabetic kidney [J] . Am J Physiol Renal Physiol , 2000, 278 :F830 - 838
39. Tanji N , Markowitz GS , Fu C , et al . Expression of advanced glycation end products and their cellular receptor RAGE in dia2betic nephropathy and nondiabetic renal disease [J ] . J Am Soc Nephrol ,2000 ,11 :1656 - 1666
40. Yamamoto Y, Kato I , Doi T, et al . Development and preven2 tion of advanced diabetic nephropathy in RAGE2overexpressing mice[J]. J Clin Invest ,2001, 108 :261 -268
41. 谭正怀、沈映军.高血糖与糖尿病肾病.中国临床药理学与治疗学,2002,7(6):565-569
42. 南征,高彦彬,钱秋海.糖尿病合并感染. 糖尿病中西医综合治疗,2002,9:351-353
43. Yoshimoto S, Sakamoto K, Wakabayashi I and Masui H. Effect of chromium administration on glucose tolerance in stroke-prone spontaneously hypertensive rats with streptozotocin-induced diabetes[J]. Metabolism ,1992 Jun,41(6) : 636–642.
44. Davis CM and Vincent JB. Chromium oligopeptide activates insulin receptor tyrosinekinase activity[J]. Biochemistry ,1997,36 (15): 4382–4385.
45. Guan X ,Matte JJ ,Ku PK .High chromium yeast supplementation improves glucose tolerance in pigs by decreasing hepatic extacction of insulin [J]. Nutr,2000,130(5):1274
46. Evans GW ,Roginksi EE ,Mertz W. Interaction of the glucose tolerance factor(GTF) with insulin[J] .Biochem Biophys Res Commun,1973,50(3):718
47. Sievenpiper JL , Enkins AL ,Whithan DL , et al . Insulin resistance : concepts , controversies , and the role of nutrition[J ] . Can J Diet Pract Res ,2002 ,63 (1) :20 - 32.
48. Katsilambros N L. Nutrition in diabetes mellitus [ J ] . Exp Clin Endocrinol Diabetes ,2001 ,109 (Suppl 2) :S250 - 258.
49. Kalkwart HJ , Bell RC, Khoury JC . Dietary fiber intakes and insulin requirement in pregnant women with type 1 diabetes[J]. Am Diet Assoc, 2001, 101 (3) : 305- 3101
50. Anonymous. American diabetes association clinical practice recommendations [J]. Diabetes Care, 2001 Jan, 24 Suppl 1: S1-133
51. ChandaliaM, Garg A, Lutjohann D, et al. Beneficial effects of high dietary fiber intake in patients with Type 2 diabetes mellitus[J]. N-Engl- J- Med, 2000 May 11, 342(19): 1392-1398.
1. 李桂芝,于道兰,刘广业.生素 E、维生素 C 对糖尿病患者心血管保护作用的研究[J]. 山东医药维,2003,43(5) :5-7
2. Stohs SJ ,Bagchi D ,Hassoun EA and Bagchi M . Oxidative mechanisms in the toxicity of chromium and cadmium ions[J]. Environ. Pathol. Toxicol. Oncol,2001, 20: 77–88.
3. Kawanishi S, Hiraku Y, Murata M and Oikawa S. The role of metals in site-specific DNA damage with reference to carcinogenesis[J]. Free Radic. Biol. Med, 2002, 32: 822–832.
4. Barceloux DG , Chromium[J]. Toxicol. Clin. Toxicol, 1999, 37: 173–194.
5. Stearns DM,Silveira SM,Wolf KK and Luke AM. Chromium(III) tris(picolinate) is mutagenic at the hypoxanthine (guanine) phosphoribosyl transferase locus in Chinese hamster ovary cells[J]. Mut. Res, 2002, 513:135–142
6. Tremblay F, Dubois MJ, Marette A. Regulation of GLUT4 traffic and function by insulin and contraction in skeletal muscle[J] . Fron tBiosci, 2003, 8:1072-1084.
7. 张娅婕, 陈秋丽, 甘振威等.Cr3 + 对糖尿病小鼠降血糖作用的研究[ J ] 吉林大学学报: 医学版, 2004 , 30 ( 3) :379-380
8. 吴蕴棠,车素萍,孙忠,等. 锌、硒、铬对糖尿病大鼠糖、脂代谢调节作用[J ] . 中国公共卫生,2004 ,20 (9) :1083 - 1085.
9. 王永芳.铬与健康研究进展[J ] 中国食品卫生杂志 ,2001 ,13(1) :452-471
10. Anderson RA ,Cheng N ,Bryden NA ,et al. Elevated intakes of supplemental chromium improves glucose and insule variables in individuals with type 2 diabetes.Diabetes, 1997 ,46:1786-1791
11. Ravina A ,Slezak L ,MirskyN ,et al. Reversal of corticosteroid-induced diabetes with supplemental chromium[J]. Diabetes Med ,1999 ,16:164-167
12. Hoenack C, Roesen P. Inhibition of angiotensin type 1 receptor prevents decline of glucose transporter (GLUT4) in diabetic rat heart. [J].Diabetes, 1996,45(suppl 1): S82-S87.
13. Marette A, AtGie C, Liu Z, et al. Differential regulation of GLUT1 and GLUT4 glucose transporters in skeletal muscle of a new model of typeII diabetes[J]. Diabetes, 1993 ,42: 1195―1201
14. Garvey WT, Maianu L, Huecksteadt TP, et al .Pretranslational suppression of a glucose transporter protein causes insulin resistance in adipocytes from patients with non-insulindependent diabetes mellitus and obesity[J]. J Clin Invest ,1991 , 87:1072―1081.
15. SUN Zhong,WU Yun-tang,CHE Su-ping,WANG Xia,GUO Gang. Study on regulation of chromium on GLUT4 expression of skeletal muscle in diabetic rats[J]. Acta Nutrimenta Sinica, 2005 ,27 (3):196-199
16. 甘振威, 张娅婕 , 刘华锋 , 李晨旭, 张静玲 , 谢林. 锌和铬对糖尿病小鼠的降血糖及抗氧化作用[J]. 吉林大学学报( 医学版),2006,32(3):410-412
17. 秦俊法. 微量元素与糖尿病[J]. 广东微量元素科学, 2000 ,7(2):1-15。
18. Shi SJ , Prenss HG, Abernethy DR , et al. Elevated blood pressure in spontaneously hypertension rats : consuming a high sucrose diet is associated with elevated angiotensin and is reversed by vanadium [J ] . Hypertens , 1997 , 15(8) : 857～862.
19. Gil J , Miralpeix M , Carreras J , et al. Insulin - like effects of vanadate on glucokinase activity and fructose 2 , 6 - bisphophate levels in the liver of diabetic rats[ J]. Biol Chem ,1988 ,263 :1868
20. Badmaev V, Prakash S and Majeed M .Vanadium: a review of its potential role in the fight against diabetes[ J]. Altern. Complem. Med ,1999 ,5:273–291.
21. Rehder D.Biological, medicinal aspects of vanadium[J]. Inorg. Chem. Commun, 2003, 6: 604–617.
22. Tsiani E and Fantus IG. Vanadium compounds. Biological actions and potential as pharmacological agents[J].Trends Endocrinol. Metab , 1997, 8: 51–58.
23. Conconi MT, DeCarlo E and Vigolo S ,et al. Effects of some vanady coordination compounds on the in vitro insulin release from rat pancreatic islets[J]. Horm. Metab. Res, 2003, 35: 402–406.
24. Cam MC, Brownsey RW and McNeill JH. Mechanisms of vanadium action: insulin-mimetic or insulin-enhancing agent[J]. Can. J. Physiol. Pharm, 2000 ,78 : 829–847
25. Brand RM and Hamel FG. Transdermally delivered peroxovanadium can lower blood glucose levels in diabetic rats[J]. Pharm ,1999 ,183:117–123
26. Cadene A, Gross R and Poucheret P et al. Vanadyl sulphate differently influencesinsulin response to glucose in isolated pancreas of normal rats after in vivo or in vitro exposure[J]. Pharm ,1996 ,318:145–151.
27. Goldfine AB, Patti ME and Zuberi L et al. Metabolic effects of vanadyl sulfate in humans with non-insulin-dependent diabetes mellitus: in vivo and in vitro studies[J]. Metabolism ,2000 ,49: 400–410
28. Shechter Y, Goldwase I, Mironchik M, et al. Historic perspective and recent developments on the insulin-like actions of vanadium; toward developing vanadium-based drugs for diabetes[J]. Coord. Chem. Rev ,2003 , 237:3–11
29. Dehghani GA, Sotoodeh M and Omrani GhR. Trophic effects of vanadium on beta-cells of STZ-induced insulin dependent diabetic rats and evidence for long term relief of diabetes mellitus[J]. Indian J. Med. Res ,1999, 110:70–75.
30. Srivastava AK. Anti-diabetic and toxic effects of vanadium compounds[J]. Mol. Cell. Biochem ,2000 ,206:177–182
31. DomingoJL. Vanadium and tungsten derivatives as antidiabetic agents: a review of their toxic effects[J]. Biol. Trace Elem. Res ,2002 ,88:97–112
32. 俞凤,叶玲霞,戚其平.血液中某些必需微量元素与高血压的关系探讨[J]. 广东微量元素科学,2005 ,12(1):23-26
33. 赵汉芬. 锌与三大物资代谢[J].微量元素与健康研究,1996 ,13(2):63
34. Sprietsma JE ,Schuitemaker GE .Diabetes can be prevented by reducing insulin production[J] .Med Hypotheses,1994 ,42(1):15-23.
35. Silvestro RA. Zinc in relat ion to diabetes and oxidative disease[J ]. J N utr, 2000, 130: 1509- 1511.
36. 李涛,黄连珍,杨雪锋. 补充锌和维生素 E 对糖尿病大鼠血一氧化氮和内皮素的影响[J] .营养学报 ,2002 ,24 (4): 416-419
37. 张桂珍,崔亚南,薄立华,等. 硒和维生素 E 对大鼠糖尿病易感性的影响[J ] . 营养学报 ,2001 ,23 (2) :110 - 113.
38. Ziegler E , Filer LJ . Present knowledge in nutrition [M] . Washing2 ton DC: International Life Science Institute ,1996: 282 - 294.
39. Sheng XQ ,Huang KX ,Xu HB. New experimental observation on therelationship of selenium and diabetes mellitus [ J ] . Biol Trace ElemRes ,2004 ,99 (3) :241 - 254.
40. Faure P. Protective effects of antioxidant micronutrients ( vitamin E ,zinc and selenium)in type 2 diabetes mellitus[J ] . Clin Chem Lab Med ,2003 ,41 (8) :995 - 998.
41. Aydin A ,Orhan H ,Sayal A ,et al. Oxidative stress and nitric oxide related parameters in type II diabetes mellitus : effects of glycemic control[J] . Clin Biochem ,2001 ,34 (1) :65 - 70.
42. 陈璐璐. 微量元素与糖尿病[J]. 中国临床营养杂志 2002, 10 (3):220-224
43. Rubenstein AH , Levin NW, Elliott GA. Manganese - induced hypoglycemia [J] . Lancet , 1962 , 2 : 1348～1351.
44. Korc M. Manganese action on pancreatic protein synthesis in normal and diabetic rats [J] . Am J Physiol , 1983 , 245 :628～634.
45. Freeland - Graves J H , Behardi F , Bales C W, et al. Metabolic balance of manganese in young men consuming diets containing five levels of dietary manganese [J] . J Nutr , 1988 , 118 : 764～773.
46. 迟锡增. 微量元素与人体健康[M] . 北京:化学工业出版社,1997:200 - 268
47. Gaede P, Poulse HE, Parving HH, et al. Double2blind, random ized study of the effect of combined treatment w ith vitam in C and vitam in E on album inuria in Type 2 diabetic patients. Diabet Med ,2001, 18: 756-760
48. Auliffe AV ,Brooks BA , Fisher EJ , et al. Administration of ascorbic acid and aldose reductase in hibitor (To lrestat) in diabetes: effect on urinary album in excretion[J]. Neph ron, 1998, 80: 277-284
49. Brusell SE ,Clermont AC ,Aiello LP ,et al .High dose vitamin E supplementation normalized retinal blood flow and creatinine clearance in patients with type 1 diabetes[J] .Diabetes Care ,1999, 22:1245-1251
50. Ohara Y, Sayegh HS, Harison DG. Regulation of endothelial constitutive nitric oxide synthase by protein k inase C[J]. Hypertension,1995, 25: 415-420
51. Faruqi R, Motte C, Dicorleto PE. Alphatocophero l in Hibits agonistinduced monocytic cell adhesion to cultured humanendo thelial cells[J]. Clin Invest, 1994, 94: 592-600
52. 葛可佑.维生素 E. 中国营养科学全书[M] ,2004 ,10:188-198
53. 韦玉和, 韦永祥, 胡惠萍. 维生素 C 对糖尿病患者血管内皮细胞的保护作用[J]. 中国综合临床杂志, 2002, 18: 59-60
54. Ting HH, Timimi FK, Boles KS, et al. Vitam in C imp roves endo thelium dependent vasodilation in patients with noninsulin dependent diabetes mellitus [J]. Ch in Invest,1996, 97: 22-28
55. Kurpakus Wheater M , Kernacki KA , Hazlett LD. Maintainingcorneal integrity how the“window”stays clear [J] . Prog His2 tochem Cytochem ,2001 ,36 (3) :185.
56. 任国峰,黄忆明,刘富强. 维生素 A 水平对孕期大鼠脂质过氧化及抗氧化系统的影响[ ] . 实用预防医学,2001 ,8 (3) :173.
57. Akerblom HK, Vaarala O, Hyoty H, et al. Environmental factors in the etiology of type 1 diabetes[J]. AM J Med Genet, 2002, 115 (1) : 18～29
58. Overbergh L, Decallonne B, WaerM, et al.Alpha,252Dihydroxyvita2 min D3 induces an autoantigen2specific T2helper 1 /T2helpe 2 immune shift in NOD mice immunized with GAD65 ( p5242p543 )[J] . Diabetes, 2000, 49 (8) :1301-1307
59. Adorini L. Tolerogenic dendrtic cells induced by vitamin D receptor lig2ands enhance regulatory T cells inhibiting autoimmune diabetes[J]. Ann N Y Acad Sci, 2003, 987: 258～261
60. AdoriniL, Penna G, Giarratana N, et al. Tolerogenic dendritic cells induced by vitamin D receptor ligands enhance regulatory T cells inhibiting allograft rejection and autoimmune diaseases[J]. Cell Biochem, 2003, 88 ( 2 ) :227～233
61. RauschFan X, Leutmezer F, Willheim M, et al. Regulation of cyto2kine p roduction in human peripheral blood mononuclear cells and allergenspe2cific the cell clones by 1 Alpha, 252dihydroxyvitamin D3[J]. Int Arch Allergy Immunol, 2002, 128 (1) : 33～41
62. Hypponen E, Laara E, Reunanen A, et al. Intake of vitamin D and risk of type 1 diabetes: a birth2cohort study[J]. Lancet, 2001, 358 ( 9292) : 1500 ～1503
2. Tiedge M, Richter T and Lenzen S. Importance of cysteine residues for the stability and catalytic activity of human pancreatic beta cell glucokinase[J].Arch Biochem Biophys, 2000, 375:251–260.
3. Bloch KO, Zemel R, Bloch OV, Grief H and Vardi P. Streptozotocin and alloxan-based selection improves toxin resistance of insulin-producing RINm cells[J]. Int J Exp Diabetes Res, 2000, 1 : 211–219.
4. Gai W, Schott-Ohly P, Gleichmann H, et al. Differential target molecules for toxicity induced by streptozotocin and alloxan in pancreatic islets of mice in vitro[J]. Exp Clin Endocrinol Diabetes ,2004, 112 : 29–37
5. Okamoto H. New aspects to the functioning and regeneration of pancreatic β-cells[J]. Adv Exp Med. Biol ,1997, 426: 301–311.
6. Sentman ML, Jonsson LM and Marklund SL. Enhanced alloxan-induced beta-cell damage and delayed recovery from hyperglycemia in mice lacking extracellular - superoxide dismutase[J]. Free Radic Biol Med, 1999, 27:790–796.
7. 张晓波,姚海涛,杨子安,等.有机铬对实验性糖尿病大鼠胰腺保护作用的观察 [J]. 黑龙江医药科学,2004 ,27(5):1-2
8. 胡晓斐,张永奎,孙文均,等 .鱼提取物(鱼酱油)对大鼠血糖及体重影响的实验研究[J].中华航海医学与高气压医学杂志,2006,13(1):23-25
9. DeFronzo RA. Diabetes ,1988, 37:667–687.
10. Weir GC, Laybutt DR, Kaneto etc. Diabetes, 2001 ,50 (Suppl.1): 154–159.
11. Rossetti L, Giaccari A and De Fronzo RA. Diabetes Care ,1990, 13: 610–630.
12. Donath MY, Gross DJ, Cerasi E and Kaiser N. Diabetes ,1999, 48: 738–744.
13. McGarry JD and Dobbins RL .Diabetologia, 1999, 42: 128–138.
14. Lorenzo A, Razzaboni B, Weir GC and Yankner BA. Nature, 1994, 368:756–760.
15. Polonsky KS, Sturis J and Bell GI. Engl N .Med, 1996, 334: 777–783.
16. Brownlee M. Nature ,2001 ,414: 813–820.
17. Robertson RP. Biol Chem, 2004, 279:42351–42354.
18. Thornalley P, Wolff S, Crabbe J and Stern A. Biochim Biophys Acta ,1984, 797:276–287.
19. 南征、高彦彬、钱秋海主编. 糖尿病中西医综合治疗, 北京:人民卫生出版社, 2002: 453
20. Clark A , Wells CA , Buley ID , et al . Islet amyloid , increased A - cells ,reduced B - cells and exocrine fibrosis : quantitative changes in the pancreas in type 2 diabetes. Diabetes Res , 1988 ,9 :151
21. Butler AE , Janson J , Ritzel R ,et al . Accelerated apoptosis overcomes in creased replication to causeβ- cell loss in diabetes in mice transgenic for h - IAPP. Diabetes , 2002 ,51 (Suppl 2) :A7
22. Butler AE , Janson J , Walter C , et al . Increased B - cell apoptosis pre2vents adaptive increase in B - cell mass in mouse model of type 2 dia2betes : evidence for role of islet amyloid formation rather than direct action of amyloid. Diabetes , 2003 , 52 :2304
23. Butler AE , Janson J , Bonner - Weir S ,et al . Beta - cell deficit and in 2 creased beta - cell apoptosis in humans with type 2 diabetes. Diabetes ,2003 , 52 :102
24. Biarnes M, Montolio M, Nacher V , et al .β- cell death and mass in syn geneically transplanted islets exposed to short - and long - term hyper2 glycemia. Diabetes , 2002 , 51 :66
25. Federici M, Hribal M, Perego L , et al . High glucose causes apoptosis in cultured human pancreatic islets of langerhans. Diabetes , 2001 , 50 :1290
26. Kaneto H, Fujii J and Myint T etal. Biochem, 1996, 320 (Pt. 3):855–863.
27. 张娅婕, 陈秋丽, 甘振威, 等 1 Cr3 + 对糖尿病小鼠降血糖作用的研究[ J ]吉林大学学报: 医学版, 2004 , 30 ( 3) :379-380
28. 吴蕴棠,车素萍,孙忠,等. 锌、硒、铬对糖尿病大鼠糖、脂代谢调节作用[J ] . 中国公共卫生,2004 ,20 (9) :1083 - 1085.
29. Monnier VM. Gerontol ,1990, 45: B105–B111.
30. Tremblay F , Dubois MJ , Marette A. Regulationof GLUT4 traffic and function by insulin and contraction in skeletal muscle[J] . Front Biosci,2003,8:1072―1084.
31. Evans JL, Goldfine ID, Maddux BA and Grodsky GM. Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes[J]. Endocr Rev ,2002, 23: 599–622.
32. Zelko IN, Mariani TJ and Folz JR. Superoxide dismutase multigene family: a comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression[J]. Free Radic Biol Med, 2002, 33:337–349.
33. Fattman CL, Schaefer LM and Oury TD. Extracellular superoxide dismutase in biology and medicine[J]. Free Radic Biol Med, 2003, 35:236–256.
34. Adachi T, Inoue M, Hara H, Maehata E and Suzuki S. Relationship of plasma extracellular-superoxide dismutase level with insulin resistance in type 2 diabetic patients[J]. Endocrinol, 2004, 181: 413–417
35. Kurien BT, Scofield RH. Free radical mediated peroxidative damage in systemic lupus erythematosu[J].Life Sci,2003,73(13):1655-1666.
36. Malarkodi KP ,Balachandar AV, Sivaprasad R,et al. Prophylactic effect of lipoic acid against adriamycin-induced peroxidative damages in rat kidney[J]. Ren Fail, 2003, 25(3):367-377.
37. Hail ED, Andrus PK, Oostveen JA, et al. Relationship of oxygen radical-induced lipid peroxidative damage to disease onset and progression in a transgenic model of familial ALS[J]. JNeurosci Res, 1998, 53(1):66-77.
38. Isono M, Mogyorosi A ,Han DC , et al . Stimulation of TGF2beta type Ⅱreceptor by high glucose in mouse mesangial cells and in diabetic kidney [J] . Am J Physiol Renal Physiol , 2000, 278 :F830 - 838
39. Tanji N , Markowitz GS , Fu C , et al . Expression of advanced glycation end products and their cellular receptor RAGE in dia2betic nephropathy and nondiabetic renal disease [J ] . J Am Soc Nephrol ,2000 ,11 :1656 - 1666
40. Yamamoto Y, Kato I , Doi T, et al . Development and preven2 tion of advanced diabetic nephropathy in RAGE2overexpressing mice[J]. J Clin Invest ,2001, 108 :261 -268
41. 谭正怀、沈映军.高血糖与糖尿病肾病.中国临床药理学与治疗学,2002,7(6):565-569
42. 南征,高彦彬,钱秋海.糖尿病合并感染. 糖尿病中西医综合治疗,2002,9:351-353
43. Yoshimoto S, Sakamoto K, Wakabayashi I and Masui H. Effect of chromium administration on glucose tolerance in stroke-prone spontaneously hypertensive rats with streptozotocin-induced diabetes[J]. Metabolism ,1992 Jun,41(6) : 636–642.
44. Davis CM and Vincent JB. Chromium oligopeptide activates insulin receptor tyrosinekinase activity[J]. Biochemistry ,1997,36 (15): 4382–4385.
45. Guan X ,Matte JJ ,Ku PK .High chromium yeast supplementation improves glucose tolerance in pigs by decreasing hepatic extacction of insulin [J]. Nutr,2000,130(5):1274
46. Evans GW ,Roginksi EE ,Mertz W. Interaction of the glucose tolerance factor(GTF) with insulin[J] .Biochem Biophys Res Commun,1973,50(3):718
47. Sievenpiper JL , Enkins AL ,Whithan DL , et al . Insulin resistance : concepts , controversies , and the role of nutrition[J ] . Can J Diet Pract Res ,2002 ,63 (1) :20 - 32.
48. Katsilambros N L. Nutrition in diabetes mellitus [ J ] . Exp Clin Endocrinol Diabetes ,2001 ,109 (Suppl 2) :S250 - 258.
49. Kalkwart HJ , Bell RC, Khoury JC . Dietary fiber intakes and insulin requirement in pregnant women with type 1 diabetes[J]. Am Diet Assoc, 2001, 101 (3) : 305- 3101
50. Anonymous. American diabetes association clinical practice recommendations [J]. Diabetes Care, 2001 Jan, 24 Suppl 1: S1-133
51. ChandaliaM, Garg A, Lutjohann D, et al. Beneficial effects of high dietary fiber intake in patients with Type 2 diabetes mellitus[J]. N-Engl- J- Med, 2000 May 11, 342(19): 1392-1398.
1. 李桂芝,于道兰,刘广业.生素 E、维生素 C 对糖尿病患者心血管保护作用的研究[J]. 山东医药维,2003,43(5) :5-7
2. Stohs SJ ,Bagchi D ,Hassoun EA and Bagchi M . Oxidative mechanisms in the toxicity of chromium and cadmium ions[J]. Environ. Pathol. Toxicol. Oncol,2001, 20: 77–88.
3. Kawanishi S, Hiraku Y, Murata M and Oikawa S. The role of metals in site-specific DNA damage with reference to carcinogenesis[J]. Free Radic. Biol. Med, 2002, 32: 822–832.
4. Barceloux DG , Chromium[J]. Toxicol. Clin. Toxicol, 1999, 37: 173–194.
5. Stearns DM,Silveira SM,Wolf KK and Luke AM. Chromium(III) tris(picolinate) is mutagenic at the hypoxanthine (guanine) phosphoribosyl transferase locus in Chinese hamster ovary cells[J]. Mut. Res, 2002, 513:135–142
6. Tremblay F, Dubois MJ, Marette A. Regulation of GLUT4 traffic and function by insulin and contraction in skeletal muscle[J] . Fron tBiosci, 2003, 8:1072-1084.
7. 张娅婕, 陈秋丽, 甘振威等.Cr3 + 对糖尿病小鼠降血糖作用的研究[ J ] 吉林大学学报: 医学版, 2004 , 30 ( 3) :379-380
8. 吴蕴棠,车素萍,孙忠,等. 锌、硒、铬对糖尿病大鼠糖、脂代谢调节作用[J ] . 中国公共卫生,2004 ,20 (9) :1083 - 1085.
9. 王永芳.铬与健康研究进展[J ] 中国食品卫生杂志 ,2001 ,13(1) :452-471
10. Anderson RA ,Cheng N ,Bryden NA ,et al. Elevated intakes of supplemental chromium improves glucose and insule variables in individuals with type 2 diabetes.Diabetes, 1997 ,46:1786-1791
11. Ravina A ,Slezak L ,MirskyN ,et al. Reversal of corticosteroid-induced diabetes with supplemental chromium[J]. Diabetes Med ,1999 ,16:164-167
12. Hoenack C, Roesen P. Inhibition of angiotensin type 1 receptor prevents decline of glucose transporter (GLUT4) in diabetic rat heart. [J].Diabetes, 1996,45(suppl 1): S82-S87.
13. Marette A, AtGie C, Liu Z, et al. Differential regulation of GLUT1 and GLUT4 glucose transporters in skeletal muscle of a new model of typeII diabetes[J]. Diabetes, 1993 ,42: 1195―1201
14. Garvey WT, Maianu L, Huecksteadt TP, et al .Pretranslational suppression of a glucose transporter protein causes insulin resistance in adipocytes from patients with non-insulindependent diabetes mellitus and obesity[J]. J Clin Invest ,1991 , 87:1072―1081.
15. SUN Zhong,WU Yun-tang,CHE Su-ping,WANG Xia,GUO Gang. Study on regulation of chromium on GLUT4 expression of skeletal muscle in diabetic rats[J]. Acta Nutrimenta Sinica, 2005 ,27 (3):196-199
16. 甘振威, 张娅婕 , 刘华锋 , 李晨旭, 张静玲 , 谢林. 锌和铬对糖尿病小鼠的降血糖及抗氧化作用[J]. 吉林大学学报( 医学版),2006,32(3):410-412
17. 秦俊法. 微量元素与糖尿病[J]. 广东微量元素科学, 2000 ,7(2):1-15。
18. Shi SJ , Prenss HG, Abernethy DR , et al. Elevated blood pressure in spontaneously hypertension rats : consuming a high sucrose diet is associated with elevated angiotensin and is reversed by vanadium [J ] . Hypertens , 1997 , 15(8) : 857～862.
19. Gil J , Miralpeix M , Carreras J , et al. Insulin - like effects of vanadate on glucokinase activity and fructose 2 , 6 - bisphophate levels in the liver of diabetic rats[ J]. Biol Chem ,1988 ,263 :1868
20. Badmaev V, Prakash S and Majeed M .Vanadium: a review of its potential role in the fight against diabetes[ J]. Altern. Complem. Med ,1999 ,5:273–291.
21. Rehder D.Biological, medicinal aspects of vanadium[J]. Inorg. Chem. Commun, 2003, 6: 604–617.
22. Tsiani E and Fantus IG. Vanadium compounds. Biological actions and potential as pharmacological agents[J].Trends Endocrinol. Metab , 1997, 8: 51–58.
23. Conconi MT, DeCarlo E and Vigolo S ,et al. Effects of some vanady coordination compounds on the in vitro insulin release from rat pancreatic islets[J]. Horm. Metab. Res, 2003, 35: 402–406.
24. Cam MC, Brownsey RW and McNeill JH. Mechanisms of vanadium action: insulin-mimetic or insulin-enhancing agent[J]. Can. J. Physiol. Pharm, 2000 ,78 : 829–847
25. Brand RM and Hamel FG. Transdermally delivered peroxovanadium can lower blood glucose levels in diabetic rats[J]. Pharm ,1999 ,183:117–123
26. Cadene A, Gross R and Poucheret P et al. Vanadyl sulphate differently influencesinsulin response to glucose in isolated pancreas of normal rats after in vivo or in vitro exposure[J]. Pharm ,1996 ,318:145–151.
27. Goldfine AB, Patti ME and Zuberi L et al. Metabolic effects of vanadyl sulfate in humans with non-insulin-dependent diabetes mellitus: in vivo and in vitro studies[J]. Metabolism ,2000 ,49: 400–410
28. Shechter Y, Goldwase I, Mironchik M, et al. Historic perspective and recent developments on the insulin-like actions of vanadium; toward developing vanadium-based drugs for diabetes[J]. Coord. Chem. Rev ,2003 , 237:3–11
29. Dehghani GA, Sotoodeh M and Omrani GhR. Trophic effects of vanadium on beta-cells of STZ-induced insulin dependent diabetic rats and evidence for long term relief of diabetes mellitus[J]. Indian J. Med. Res ,1999, 110:70–75.
30. Srivastava AK. Anti-diabetic and toxic effects of vanadium compounds[J]. Mol. Cell. Biochem ,2000 ,206:177–182
31. DomingoJL. Vanadium and tungsten derivatives as antidiabetic agents: a review of their toxic effects[J]. Biol. Trace Elem. Res ,2002 ,88:97–112
32. 俞凤,叶玲霞,戚其平.血液中某些必需微量元素与高血压的关系探讨[J]. 广东微量元素科学,2005 ,12(1):23-26
33. 赵汉芬. 锌与三大物资代谢[J].微量元素与健康研究,1996 ,13(2):63
34. Sprietsma JE ,Schuitemaker GE .Diabetes can be prevented by reducing insulin production[J] .Med Hypotheses,1994 ,42(1):15-23.
35. Silvestro RA. Zinc in relat ion to diabetes and oxidative disease[J ]. J N utr, 2000, 130: 1509- 1511.
36. 李涛,黄连珍,杨雪锋. 补充锌和维生素 E 对糖尿病大鼠血一氧化氮和内皮素的影响[J] .营养学报 ,2002 ,24 (4): 416-419
37. 张桂珍,崔亚南,薄立华,等. 硒和维生素 E 对大鼠糖尿病易感性的影响[J ] . 营养学报 ,2001 ,23 (2) :110 - 113.
38. Ziegler E , Filer LJ . Present knowledge in nutrition [M] . Washing2 ton DC: International Life Science Institute ,1996: 282 - 294.
39. Sheng XQ ,Huang KX ,Xu HB. New experimental observation on therelationship of selenium and diabetes mellitus [ J ] . Biol Trace ElemRes ,2004 ,99 (3) :241 - 254.
40. Faure P. Protective effects of antioxidant micronutrients ( vitamin E ,zinc and selenium)in type 2 diabetes mellitus[J ] . Clin Chem Lab Med ,2003 ,41 (8) :995 - 998.
41. Aydin A ,Orhan H ,Sayal A ,et al. Oxidative stress and nitric oxide related parameters in type II diabetes mellitus : effects of glycemic control[J] . Clin Biochem ,2001 ,34 (1) :65 - 70.
42. 陈璐璐. 微量元素与糖尿病[J]. 中国临床营养杂志 2002, 10 (3):220-224
43. Rubenstein AH , Levin NW, Elliott GA. Manganese - induced hypoglycemia [J] . Lancet , 1962 , 2 : 1348～1351.
44. Korc M. Manganese action on pancreatic protein synthesis in normal and diabetic rats [J] . Am J Physiol , 1983 , 245 :628～634.
45. Freeland - Graves J H , Behardi F , Bales C W, et al. Metabolic balance of manganese in young men consuming diets containing five levels of dietary manganese [J] . J Nutr , 1988 , 118 : 764～773.
46. 迟锡增. 微量元素与人体健康[M] . 北京:化学工业出版社,1997:200 - 268
47. Gaede P, Poulse HE, Parving HH, et al. Double2blind, random ized study of the effect of combined treatment w ith vitam in C and vitam in E on album inuria in Type 2 diabetic patients. Diabet Med ,2001, 18: 756-760
48. Auliffe AV ,Brooks BA , Fisher EJ , et al. Administration of ascorbic acid and aldose reductase in hibitor (To lrestat) in diabetes: effect on urinary album in excretion[J]. Neph ron, 1998, 80: 277-284
49. Brusell SE ,Clermont AC ,Aiello LP ,et al .High dose vitamin E supplementation normalized retinal blood flow and creatinine clearance in patients with type 1 diabetes[J] .Diabetes Care ,1999, 22:1245-1251
50. Ohara Y, Sayegh HS, Harison DG. Regulation of endothelial constitutive nitric oxide synthase by protein k inase C[J]. Hypertension,1995, 25: 415-420
51. Faruqi R, Motte C, Dicorleto PE. Alphatocophero l in Hibits agonistinduced monocytic cell adhesion to cultured humanendo thelial cells[J]. Clin Invest, 1994, 94: 592-600
52. 葛可佑.维生素 E. 中国营养科学全书[M] ,2004 ,10:188-198
53. 韦玉和, 韦永祥, 胡惠萍. 维生素 C 对糖尿病患者血管内皮细胞的保护作用[J]. 中国综合临床杂志, 2002, 18: 59-60
54. Ting HH, Timimi FK, Boles KS, et al. Vitam in C imp roves endo thelium dependent vasodilation in patients with noninsulin dependent diabetes mellitus [J]. Ch in Invest,1996, 97: 22-28
55. Kurpakus Wheater M , Kernacki KA , Hazlett LD. Maintainingcorneal integrity how the“window”stays clear [J] . Prog His2 tochem Cytochem ,2001 ,36 (3) :185.
56. 任国峰,黄忆明,刘富强. 维生素 A 水平对孕期大鼠脂质过氧化及抗氧化系统的影响[ ] . 实用预防医学,2001 ,8 (3) :173.
57. Akerblom HK, Vaarala O, Hyoty H, et al. Environmental factors in the etiology of type 1 diabetes[J]. AM J Med Genet, 2002, 115 (1) : 18～29
58. Overbergh L, Decallonne B, WaerM, et al.Alpha,252Dihydroxyvita2 min D3 induces an autoantigen2specific T2helper 1 /T2helpe 2 immune shift in NOD mice immunized with GAD65 ( p5242p543 )[J] . Diabetes, 2000, 49 (8) :1301-1307
59. Adorini L. Tolerogenic dendrtic cells induced by vitamin D receptor lig2ands enhance regulatory T cells inhibiting autoimmune diabetes[J]. Ann N Y Acad Sci, 2003, 987: 258～261
60. AdoriniL, Penna G, Giarratana N, et al. Tolerogenic dendritic cells induced by vitamin D receptor ligands enhance regulatory T cells inhibiting allograft rejection and autoimmune diaseases[J]. Cell Biochem, 2003, 88 ( 2 ) :227～233
61. RauschFan X, Leutmezer F, Willheim M, et al. Regulation of cyto2kine p roduction in human peripheral blood mononuclear cells and allergenspe2cific the cell clones by 1 Alpha, 252dihydroxyvitamin D3[J]. Int Arch Allergy Immunol, 2002, 128 (1) : 33～41
62. Hypponen E, Laara E, Reunanen A, et al. Intake of vitamin D and risk of type 1 diabetes: a birth2cohort study[J]. Lancet, 2001, 358 ( 9292) : 1500 ～1503