清润方对糖尿病的治疗作用及其机制的实验研究
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摘要
随着我国社会经济的发展,人民生活方式改变,体力活动减少以及肥胖的增加等因素导致糖尿病患病人数骤增。2010年《新英格兰医学杂志》发文指出中国20岁以上成人糖尿病的患病率高达9.7%,糖尿病前期的患病率达15.5%,已成为严重危害群众健康的公共健康问题。中医药一直在糖尿病的防治方面做着积极的努力,中国中医科学院广安门医院林兰教授历经40余年临床与科研创立的糖尿病中医三型辨证理论(阴虚热盛、气阴两虚、阴阳两虚),客观的反应了糖尿病中医证型的动态演变规律,有效地指导了糖尿病中医治疗的临床实践。清润方(由知母、黄柏、酒大黄等组成)专为糖尿病早期的阴虚热盛型而设,以滋阴清热法治疗糖尿病早期阴虚热盛患者临床疗效显著。本研究以清润方为研究对象,利用糖尿病模型大鼠观察了不同剂量清润方对糖尿病大鼠糖代谢的影响,并从脂代谢、肝脏炎症与氧化应激、骨骼肌糖代谢关键基因mRNA表达等方面探讨了其发挥作用可能的机制,以期为清润方在糖尿病治疗中的临床应用提供理论依据与数据支撑。
1.文献综述：本部分分别从糖尿病与胰岛素抵抗的关系、胰岛素抵抗相关机制(脂代谢紊乱、炎症与氧化应激、骨骼肌胰岛素抵抗)的研究现状、清润方组方原理与方中单味中药现代药理研究及中药治疗糖尿病研究进展等方面对本研究依托的研究基础进行了综述。
2.实验研究：包括以下四个基础实验。
2.1清润方对实验性糖尿病大鼠糖代谢及胰腺病理学形态的影响
目的：观察清润方对糖尿病大鼠糖代谢及胰腺病理学形态的影响。方法：利用链脲佐菌素(streptozotocin, STZ)联合高糖高脂饲料的方法建立糖尿病大鼠模型。将实验动物分为空白对照组、模型组、清润方小、中、大剂量组(0.5,1,2g·kg·1·d-1)、二甲双胍组(150mg·kg-1·d-1),各治疗组分别灌胃给予相应药物治疗。干预治疗4周后检测各组大鼠体重、空腹血糖(FBG)、血清胰岛素水平、口服糖耐量曲线下面积(AUC),计算胰岛素敏感指数(ISI)与胰岛素抵抗指数(IRI),光镜、电镜下观察胰腺病理形态学改变。结果：清润方中、大剂量组和二甲双胍组较模型组FBG降低(P<0.05)；清润方各剂量组和二甲双胍组AUC均有不同程度的降低(P<0.05)；清润方中、小剂量组和二甲双胍组较模型组ISI升高,胰岛素抵抗指数IRI降低(P<0.05)；清润方中、小剂量组和二甲双胍组肝糖原含量升高(P<0.05)；光镜下显示清润方中、大剂量组及二甲双胍组胰岛数目的减少有所好转,虽大部分胰岛体积明显缩小,仍可见较大的胰岛；电镜下清润方各剂量组及二甲双胍组胰岛β细胞内分泌颗粒较模型组增多。结论：清润方有降低糖尿病大鼠血糖的作用,其机制与改善糖尿病大鼠的胰岛素抵抗,保护胰岛功能,促进糖尿病大鼠对葡萄糖的利用及肝糖原的合成有关。
2.2清润方对实验性糖尿病大鼠脂代谢及肝脏病理学形态的影响
目的：观察清润方对糖尿病大鼠脂代谢及肝脏病理形态学的影响,探讨其在脂代谢方而改善胰岛素抵抗的作用。方法：各组大鼠药物干预4周后取大鼠血清及肝脏组织,全血自动生化分析仪COD-PAP法测定总胆固醇(TCHO), GPO-PAP法测定三酰甘油(TG),清除法测定高密度脂蛋白胆固醇(HDL-C)与低密度脂蛋白胆固醇(LDL-C);铜显色法测定血清游离脂肪酸水平；光镜下观察肝脏病理形态学改变。结果：与模型组比较,清润方中、大剂量组糖尿病大鼠的TCHO、LDL-C水平降低(P<0.05),清润方各剂量组和二甲双胍组糖尿病大鼠的HDL-C水平升高(P<0.05),TG水平有降低趋势但无统计学差异；清润方中、小剂量组和二甲双胍组大鼠血清游离脂肪酸水平降低(P<0.05)；光镜下显示清润方中、大剂量组和二甲双胍组对肝脏的脂肪沉积有改善作用。结论：清润方有改善糖尿病大鼠脂代谢紊乱,抑制脂肪在肝脏沉积的作用。清润方对糖尿病大鼠“脂毒性”的改善是其发挥抗糖尿病大鼠胰岛素抵抗的机制之一。
2.3清润方对实验性糖尿病大鼠肝脏炎症及氧化应激的影响
目的：观察清润方对糖尿病大鼠肝脏炎症反应及氧化应激的影响,探讨其对肝脏胰岛素抵抗的作用机制。方法：各组大鼠药物干预4周后取大鼠肝脏组织,酶联免疫吸附法(ELISA)法测定肝脏组织肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)含量；比色法测定肝脏组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性。结果：与模型组比较,清润方各剂量组肝组织TNF-α水平降低(P<0.05),清润方小、大剂量组肝组织IL-6水平降低(P<0.05),清润方中剂量组肝组织IL-6水平有降低趋势,但无统计学差异；清润方中、小剂量组及二甲双胍组肝脏组织MDA含量降低(P<0.05),清润方中、大剂量组肝脏组织SOD活性提高。结论：清润方有降低肝脏组织炎症因子含量,促进氧自由基清除,减轻组织损伤的作用。控制慢性炎症和减少氧化应激的损伤是清润方改善肝脏胰岛素抵抗的重要机制。
2.4清润方对实验性糖尿病大鼠骨骼肌GLUT-4、INSR mRNA表达的影响
目的：观察清润方对糖尿病大鼠骨骼肌GLUT-4、INSR mRNA表达的影响,探讨其对外周胰岛素抵抗中骨骼肌胰岛素抵抗的作用机制。方法：各组大鼠药物干预4周后无菌条件下,取左后腿骨骼肌组织,利用聚合酶链式反应(PCR)检测骨骼肌组织GLUT-4、INSR mRNA的表达。结果：与模型组相比,清润方中、大剂量组和二甲双胍组GLUT-4mRNA表达升高(P<0.05),INSR mRNA表达虽有升高趋势,但无统计学差异。结论：清润方上调糖尿病大鼠骨骼肌GLUT-4mRNA表达,促进糖尿病大鼠骨骼肌对葡萄糖的利用,是其发挥改善糖尿病大鼠外周组织骨骼肌胰岛素抵抗的作用机制之一。
综上所述,本研究显示清润方有降低实验性糖尿病大鼠血糖、改善胰岛素抵抗、保护胰岛功能的作用,其发挥作用的机制可能与改善实验性糖尿病大鼠的脂代谢,减轻脂毒性；控制肝脏慢性炎症和减少氧化应激的损伤改善肝脏胰岛素抵抗；上调骨骼肌组织GLUT-4mRNA表达,促进糖尿病大鼠骨骼肌对葡萄糖的利用,改善骨骼肌胰岛素抵抗有关。为清润方在糖尿病“三型辨证”中阴虚热盛型患者中的使用提供了理论依据与数据支撑。
There is an increasing number of populations suffering from diabetes due to the china's social-economy development, people's life style changes, and decreasing physical activity and increasing number of obesity people. According to the article published in New England Medicine2010, it was estimated that the incidence of over20year old diabetes sufferer is about9.7%, for pre-diabetes the number is15.5%, and now it has been a severe public health problem. Chinese medicine plays a critical role in diabetes prevention and treatment, for example, professor Lin Lan with over40years of clinical expertise and researches in Guang' anmen Hospital Affiliated to China Academy of Chinese Medical Sciences, created the theory of three syndromes for diabetes in Chinese medicine, including yin deficiency due to excessive heat, qi&yin deficiency, and yin&yang deficiency, which summarized the dynamic alteration laws of diabetes, benefits clinical practice of diabetes in Chinese medicine. Qingrun formula (composed of huangbai, dahuang liquid soaked, zhimu) is specialized for diabetes early stage, the principle of nourishing yin and clearing heat works well for the diabetes syndrome of yin deficiency due to excessive heat. This study mainly focus on Qingrun formula's potential mechanism of lipid metabolism, liver inflammation, and oxidative stress, key gene mRNA expression on skeletal muscle metabolism, seeking theoretical evidence and database for clinical practice.
1. Reviews:several parts will be discussed, including the relation between diabetes and insulin resistance, present research on mechanism of insulin resistance (disorder of lipid metabolism, inflammation and oxidative stress, insulin resistance in skeletal muscle), composing principle of Qingrun formula, and s modern study of single herb in the formula, and Chinese medicine treatment for diabetes.
2. Experimental trials:four basic experiments are referred.
2.1Qingrun formula's effect on glucose metabolism and pancreatic pathology&morphology in experimental diabetes mice.
Objective:to observe Qingrun formula's effect on glucose metabolism and pancreatic pathology&morphology in experimental diabetes mice. Method: diabetes mice models were created by combination of streptozotocin and high sugar&lipid feeding. Experimental animals were divided into6groups, including contrast, model, Qingrun formula small, moderate, and high dosage (respective0.5,1,2g·kg-1·d-1), and Metformin groups; all groups were given medication by gastric lavage. With4weeks intervention, the mice were measured weight, fasting blood glucose, insulin level in blood, AUC, ISI, insulin resistance index, changes of pancreatic pathology&morphology in the optical microscope and electron microscope. Results:FBG was lower in Qingrun formula's moderate&high dosage and Metformin groups than model group (P<0.05); AUC in all three Qingrun formula and Metformin groups decreased individually (P<0.05); ISI in Qingrun formula's moderate&small dosage and Metformin groups were higher than model group, meantime, IRI is decreased (P<0.05); hepatic glycogen were increased in Qingrun formula's moderate&small dosage and Metformin group (P<0.05); in optical microscope, pancreatic islet account decreasing was improved in Qingrun formula's moderate&high dosage and Metformin groups, although partial islet size became smaller, occasionally larger ones is seen; in electron microscope, endocrine granule secreted by pancreatic islet (3-cell increased in all three Qingrun formula's and Metformin groups was much more than model group. Consults:Qingrun formula has good effects on lowering blood glucose in mice, the mechanism maybe referred to insulin resistance, protection of pancreatic islet, and improvement of glucose utilization and hepatic glycogen synthesis.
2.2Qingrun formula's effect on lipid metabolism and liver pathology and morphology in experimental diabetes mice
Objective:to observe Qingrun formula's effect on lipid metabolism and liver pathology and morphology in experimental diabetes mice, and discuss the function of improving insulin resistance in lipid mechanism. Method:with4weeks intervention, the mice were taken blood test, liver tissue test, TCHO by blood automatic biochemical analyzer COD-PAP, TG test by GPO-PAP, HDL-C&LDL-C measured by elimination method; Serum free fatty acid was measured with cooper color; liver tissue pathology and morphology changes in optical microscope was observed. Results:in comparison to model group, TCHO、LDL-C of diabetes mice lowered in Qingrun formulas moderate&high dosage groups (P<0.05); HDL-C increased in all three Qingrun formula and Metformin groups (P<0.05), TG showed decreased tendency without significance; Serum free fatty acid decreased in Qingrun formula's moderate&small dosage and Metformin groups (P<0.05); in optical microscope, liver fat deposition were improved in Qingrun formula's moderate&high dosage and Metformin groups. Consults:Qingrun formula has good effects on lipid metabolism and inhabiting liver fat deposition in diabetes mice. Qingrun formula's effect on lipid toxicity in diabetes mice related to the mechanism of insulin resistance.
2.3Qingrun formula's effect on liver inflammation and oxidative stress in experimental diabetes mice
Objective:to observe Qingrun formula's effect on liver inflammation and oxidative stress in experimental diabetes mice, and discuss the mechanism. Method: with4weeks intervention, liver tissue were taken out from mice, TNF-α and IL-6were measured by ELISA, MDA content and SOD activity of liver tissue were tested with colorimetry. Results:in comparison to model group, TNF-α lowered in all three Qingrun formula groups (P<0.05); IL-6of liver tissue decreased in Qingrun formula's small&high dosage groups (P<0.05), slight decreased tendency in Qingrun formula moderate dosage without significance; MDA in liver tissue decreased in Qingrun formula's moderate&small dosage and Metformin groups (P <0.05), meantime, SOD increased in Qingrun formula moderate&high dosage groups. Consults:Qingrun formula has good effects on lowering inflammatory factors in liver tissue by improve Oxygen free radical scavenging to decrease tissue damage. Controlling chronic inflammation and oxidative stress damage account for the mechanism of Qingrun formula in improving insulin resistance.
2.4Qingrun formula's effects on skeletal muscle GLUT-4、INSR mRNA expression in experimental diabetes mice
Objective:Qingrun formula's effects on skeletal muscle GLUT-4、INSR mRNA expression in experimental diabetes mice, and discuss the function of insulin resistance in skeletal muscle. Method:with4weeks intervention, skeletal muscle of left posterior legs were taken out in sterile conditions to exam GLUT-4、INSR mRNA expression by PCR. Results:in comparison to model group, GLUT-4mRNA expression increased in Qingrun formula's moderate&high dosage and Metformin groups (P<0.05), INSR mRNA expression increased without significance. Consults: Qingrun formula has a role of upgrading skeletal muscle GLUT-4mRNA expression, improving glucose utilization in skeletal muscle, which is related to the mechanism of insulin resistance in diabetes mice.
Above mentioned, this study shows that Qingrun formula has good effects on lowering blood glucose, improving insulin resistance, and protecting pancreatic islet, which the functions relate to promote lipid metabolism, and decrease lipid toxicity; controlling liver chronic inflammation and decreasing oxidative stress damage to improve liver pancreatic islet insulin resistance; promoting glucose utilization in skeletal muscle to improve insulin resistance. Besides, Qingrun formula provided theoretical evidence and database for Chinese medicine application in yin deficiency due to excessive heat in three syndromes.
1.文献综述：本部分分别从糖尿病与胰岛素抵抗的关系、胰岛素抵抗相关机制(脂代谢紊乱、炎症与氧化应激、骨骼肌胰岛素抵抗)的研究现状、清润方组方原理与方中单味中药现代药理研究及中药治疗糖尿病研究进展等方面对本研究依托的研究基础进行了综述。
2.实验研究：包括以下四个基础实验。
2.1清润方对实验性糖尿病大鼠糖代谢及胰腺病理学形态的影响
目的：观察清润方对糖尿病大鼠糖代谢及胰腺病理学形态的影响。方法：利用链脲佐菌素(streptozotocin, STZ)联合高糖高脂饲料的方法建立糖尿病大鼠模型。将实验动物分为空白对照组、模型组、清润方小、中、大剂量组(0.5,1,2g·kg·1·d-1)、二甲双胍组(150mg·kg-1·d-1),各治疗组分别灌胃给予相应药物治疗。干预治疗4周后检测各组大鼠体重、空腹血糖(FBG)、血清胰岛素水平、口服糖耐量曲线下面积(AUC),计算胰岛素敏感指数(ISI)与胰岛素抵抗指数(IRI),光镜、电镜下观察胰腺病理形态学改变。结果：清润方中、大剂量组和二甲双胍组较模型组FBG降低(P<0.05)；清润方各剂量组和二甲双胍组AUC均有不同程度的降低(P<0.05)；清润方中、小剂量组和二甲双胍组较模型组ISI升高,胰岛素抵抗指数IRI降低(P<0.05)；清润方中、小剂量组和二甲双胍组肝糖原含量升高(P<0.05)；光镜下显示清润方中、大剂量组及二甲双胍组胰岛数目的减少有所好转,虽大部分胰岛体积明显缩小,仍可见较大的胰岛；电镜下清润方各剂量组及二甲双胍组胰岛β细胞内分泌颗粒较模型组增多。结论：清润方有降低糖尿病大鼠血糖的作用,其机制与改善糖尿病大鼠的胰岛素抵抗,保护胰岛功能,促进糖尿病大鼠对葡萄糖的利用及肝糖原的合成有关。
2.2清润方对实验性糖尿病大鼠脂代谢及肝脏病理学形态的影响
目的：观察清润方对糖尿病大鼠脂代谢及肝脏病理形态学的影响,探讨其在脂代谢方而改善胰岛素抵抗的作用。方法：各组大鼠药物干预4周后取大鼠血清及肝脏组织,全血自动生化分析仪COD-PAP法测定总胆固醇(TCHO), GPO-PAP法测定三酰甘油(TG),清除法测定高密度脂蛋白胆固醇(HDL-C)与低密度脂蛋白胆固醇(LDL-C);铜显色法测定血清游离脂肪酸水平；光镜下观察肝脏病理形态学改变。结果：与模型组比较,清润方中、大剂量组糖尿病大鼠的TCHO、LDL-C水平降低(P<0.05),清润方各剂量组和二甲双胍组糖尿病大鼠的HDL-C水平升高(P<0.05),TG水平有降低趋势但无统计学差异；清润方中、小剂量组和二甲双胍组大鼠血清游离脂肪酸水平降低(P<0.05)；光镜下显示清润方中、大剂量组和二甲双胍组对肝脏的脂肪沉积有改善作用。结论：清润方有改善糖尿病大鼠脂代谢紊乱,抑制脂肪在肝脏沉积的作用。清润方对糖尿病大鼠“脂毒性”的改善是其发挥抗糖尿病大鼠胰岛素抵抗的机制之一。
2.3清润方对实验性糖尿病大鼠肝脏炎症及氧化应激的影响
目的：观察清润方对糖尿病大鼠肝脏炎症反应及氧化应激的影响,探讨其对肝脏胰岛素抵抗的作用机制。方法：各组大鼠药物干预4周后取大鼠肝脏组织,酶联免疫吸附法(ELISA)法测定肝脏组织肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)含量；比色法测定肝脏组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性。结果：与模型组比较,清润方各剂量组肝组织TNF-α水平降低(P<0.05),清润方小、大剂量组肝组织IL-6水平降低(P<0.05),清润方中剂量组肝组织IL-6水平有降低趋势,但无统计学差异；清润方中、小剂量组及二甲双胍组肝脏组织MDA含量降低(P<0.05),清润方中、大剂量组肝脏组织SOD活性提高。结论：清润方有降低肝脏组织炎症因子含量,促进氧自由基清除,减轻组织损伤的作用。控制慢性炎症和减少氧化应激的损伤是清润方改善肝脏胰岛素抵抗的重要机制。
2.4清润方对实验性糖尿病大鼠骨骼肌GLUT-4、INSR mRNA表达的影响
目的：观察清润方对糖尿病大鼠骨骼肌GLUT-4、INSR mRNA表达的影响,探讨其对外周胰岛素抵抗中骨骼肌胰岛素抵抗的作用机制。方法：各组大鼠药物干预4周后无菌条件下,取左后腿骨骼肌组织,利用聚合酶链式反应(PCR)检测骨骼肌组织GLUT-4、INSR mRNA的表达。结果：与模型组相比,清润方中、大剂量组和二甲双胍组GLUT-4mRNA表达升高(P<0.05),INSR mRNA表达虽有升高趋势,但无统计学差异。结论：清润方上调糖尿病大鼠骨骼肌GLUT-4mRNA表达,促进糖尿病大鼠骨骼肌对葡萄糖的利用,是其发挥改善糖尿病大鼠外周组织骨骼肌胰岛素抵抗的作用机制之一。
综上所述,本研究显示清润方有降低实验性糖尿病大鼠血糖、改善胰岛素抵抗、保护胰岛功能的作用,其发挥作用的机制可能与改善实验性糖尿病大鼠的脂代谢,减轻脂毒性；控制肝脏慢性炎症和减少氧化应激的损伤改善肝脏胰岛素抵抗；上调骨骼肌组织GLUT-4mRNA表达,促进糖尿病大鼠骨骼肌对葡萄糖的利用,改善骨骼肌胰岛素抵抗有关。为清润方在糖尿病“三型辨证”中阴虚热盛型患者中的使用提供了理论依据与数据支撑。
There is an increasing number of populations suffering from diabetes due to the china's social-economy development, people's life style changes, and decreasing physical activity and increasing number of obesity people. According to the article published in New England Medicine2010, it was estimated that the incidence of over20year old diabetes sufferer is about9.7%, for pre-diabetes the number is15.5%, and now it has been a severe public health problem. Chinese medicine plays a critical role in diabetes prevention and treatment, for example, professor Lin Lan with over40years of clinical expertise and researches in Guang' anmen Hospital Affiliated to China Academy of Chinese Medical Sciences, created the theory of three syndromes for diabetes in Chinese medicine, including yin deficiency due to excessive heat, qi&yin deficiency, and yin&yang deficiency, which summarized the dynamic alteration laws of diabetes, benefits clinical practice of diabetes in Chinese medicine. Qingrun formula (composed of huangbai, dahuang liquid soaked, zhimu) is specialized for diabetes early stage, the principle of nourishing yin and clearing heat works well for the diabetes syndrome of yin deficiency due to excessive heat. This study mainly focus on Qingrun formula's potential mechanism of lipid metabolism, liver inflammation, and oxidative stress, key gene mRNA expression on skeletal muscle metabolism, seeking theoretical evidence and database for clinical practice.
1. Reviews:several parts will be discussed, including the relation between diabetes and insulin resistance, present research on mechanism of insulin resistance (disorder of lipid metabolism, inflammation and oxidative stress, insulin resistance in skeletal muscle), composing principle of Qingrun formula, and s modern study of single herb in the formula, and Chinese medicine treatment for diabetes.
2. Experimental trials:four basic experiments are referred.
2.1Qingrun formula's effect on glucose metabolism and pancreatic pathology&morphology in experimental diabetes mice.
Objective:to observe Qingrun formula's effect on glucose metabolism and pancreatic pathology&morphology in experimental diabetes mice. Method: diabetes mice models were created by combination of streptozotocin and high sugar&lipid feeding. Experimental animals were divided into6groups, including contrast, model, Qingrun formula small, moderate, and high dosage (respective0.5,1,2g·kg-1·d-1), and Metformin groups; all groups were given medication by gastric lavage. With4weeks intervention, the mice were measured weight, fasting blood glucose, insulin level in blood, AUC, ISI, insulin resistance index, changes of pancreatic pathology&morphology in the optical microscope and electron microscope. Results:FBG was lower in Qingrun formula's moderate&high dosage and Metformin groups than model group (P<0.05); AUC in all three Qingrun formula and Metformin groups decreased individually (P<0.05); ISI in Qingrun formula's moderate&small dosage and Metformin groups were higher than model group, meantime, IRI is decreased (P<0.05); hepatic glycogen were increased in Qingrun formula's moderate&small dosage and Metformin group (P<0.05); in optical microscope, pancreatic islet account decreasing was improved in Qingrun formula's moderate&high dosage and Metformin groups, although partial islet size became smaller, occasionally larger ones is seen; in electron microscope, endocrine granule secreted by pancreatic islet (3-cell increased in all three Qingrun formula's and Metformin groups was much more than model group. Consults:Qingrun formula has good effects on lowering blood glucose in mice, the mechanism maybe referred to insulin resistance, protection of pancreatic islet, and improvement of glucose utilization and hepatic glycogen synthesis.
2.2Qingrun formula's effect on lipid metabolism and liver pathology and morphology in experimental diabetes mice
Objective:to observe Qingrun formula's effect on lipid metabolism and liver pathology and morphology in experimental diabetes mice, and discuss the function of improving insulin resistance in lipid mechanism. Method:with4weeks intervention, the mice were taken blood test, liver tissue test, TCHO by blood automatic biochemical analyzer COD-PAP, TG test by GPO-PAP, HDL-C&LDL-C measured by elimination method; Serum free fatty acid was measured with cooper color; liver tissue pathology and morphology changes in optical microscope was observed. Results:in comparison to model group, TCHO、LDL-C of diabetes mice lowered in Qingrun formulas moderate&high dosage groups (P<0.05); HDL-C increased in all three Qingrun formula and Metformin groups (P<0.05), TG showed decreased tendency without significance; Serum free fatty acid decreased in Qingrun formula's moderate&small dosage and Metformin groups (P<0.05); in optical microscope, liver fat deposition were improved in Qingrun formula's moderate&high dosage and Metformin groups. Consults:Qingrun formula has good effects on lipid metabolism and inhabiting liver fat deposition in diabetes mice. Qingrun formula's effect on lipid toxicity in diabetes mice related to the mechanism of insulin resistance.
2.3Qingrun formula's effect on liver inflammation and oxidative stress in experimental diabetes mice
Objective:to observe Qingrun formula's effect on liver inflammation and oxidative stress in experimental diabetes mice, and discuss the mechanism. Method: with4weeks intervention, liver tissue were taken out from mice, TNF-α and IL-6were measured by ELISA, MDA content and SOD activity of liver tissue were tested with colorimetry. Results:in comparison to model group, TNF-α lowered in all three Qingrun formula groups (P<0.05); IL-6of liver tissue decreased in Qingrun formula's small&high dosage groups (P<0.05), slight decreased tendency in Qingrun formula moderate dosage without significance; MDA in liver tissue decreased in Qingrun formula's moderate&small dosage and Metformin groups (P <0.05), meantime, SOD increased in Qingrun formula moderate&high dosage groups. Consults:Qingrun formula has good effects on lowering inflammatory factors in liver tissue by improve Oxygen free radical scavenging to decrease tissue damage. Controlling chronic inflammation and oxidative stress damage account for the mechanism of Qingrun formula in improving insulin resistance.
2.4Qingrun formula's effects on skeletal muscle GLUT-4、INSR mRNA expression in experimental diabetes mice
Objective:Qingrun formula's effects on skeletal muscle GLUT-4、INSR mRNA expression in experimental diabetes mice, and discuss the function of insulin resistance in skeletal muscle. Method:with4weeks intervention, skeletal muscle of left posterior legs were taken out in sterile conditions to exam GLUT-4、INSR mRNA expression by PCR. Results:in comparison to model group, GLUT-4mRNA expression increased in Qingrun formula's moderate&high dosage and Metformin groups (P<0.05), INSR mRNA expression increased without significance. Consults: Qingrun formula has a role of upgrading skeletal muscle GLUT-4mRNA expression, improving glucose utilization in skeletal muscle, which is related to the mechanism of insulin resistance in diabetes mice.
Above mentioned, this study shows that Qingrun formula has good effects on lowering blood glucose, improving insulin resistance, and protecting pancreatic islet, which the functions relate to promote lipid metabolism, and decrease lipid toxicity; controlling liver chronic inflammation and decreasing oxidative stress damage to improve liver pancreatic islet insulin resistance; promoting glucose utilization in skeletal muscle to improve insulin resistance. Besides, Qingrun formula provided theoretical evidence and database for Chinese medicine application in yin deficiency due to excessive heat in three syndromes.
引文
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