脾虚证2型糖尿病大鼠胃黏膜损伤模型的建立及评价
|
摘要
【目的】规范脾虚证2型糖尿病大鼠胃黏膜损伤模型的造模及评价。【方法】选用雄性SD大鼠随机分为正常组和模型组,采用高脂高糖饲料喂养和小剂量链脲佐菌素(STZ)腹腔注射的方法复制2型糖尿病大鼠模型,再结合体积分数30%乙醇灌胃的方法进一步诱发2型糖尿病胃黏膜损伤大鼠模型,检测2组大鼠血清空腹血糖(FBG)、空腹血清胰岛素(FINS)、超氧化物歧化酶(SOD)、丙二醛(MDA)和一氧化氮(NO)水平,并取胃、胰腺组织作苏木精—伊红(HE)染色观察,使用现代医学评价方法,计量化中医四诊信息并作统计学处理,对模型大鼠的证候属性及演变进行识别。【结果】2型糖尿病胃黏膜损伤大鼠模型的成模率为75%,第10~16周模型组大鼠显示脾虚证特征;模型组大鼠持续消瘦,抓力值降低,旷场活动减弱,舌质变淡,爪和尾的色度值(r值)降低,排便次数增加,粪便变软或便溏,有78%的模型大鼠出现糖尿病疾病特征、气虚证1项和脾的组织定位体征1项以上的指征。【结论】本研究成功诱发2型糖尿病胃黏膜损伤大鼠模型,建模后第10~16周为2型糖尿病胃黏膜损伤大鼠脾虚证的时间窗。
Objective To standardize the modeling and evaluation of type 2 diabetic gastric mucosal injury rat model of spleen deficiency. Methods Male SD rats were randomly divided into normal group and model group,type 2 diabetic rat model with gastric mucosal injury was established by the injection of small-dose streptozotocin(STZ) combined with feeding of high fat and sugar forage and gastric gavage of 30% ethanol solution. After the modeling, we examined the levels of fasting blood glucose( FBG), fasting insulin( FINS), superoxide dismutase(SOD), malondialdehyde(MDA) and nitric oxide(NO) in the rat serum, and gastric and pancreatic tissue pathology was evaluated after HE staining. Modern medical evaluation method was used for the statistics of information collected by four diagnostic methods of traditional Chinese medicine( TCM), and for the identification of properties and evolution of TCM syndromes. Results Type 2 diabetic gastric mucosal injury rat model had been established in 75% of the rats. And 10-16 weeks after the modeling, the model rats had the syndrome of spleen deficiency, showing as persistent emaciation, decreased grasping force, weak activities in open field test, pale tongue nature, decreased r value of the claw and tail, increased defecation frequency,softening of loose stool. Of all the model rats, 78% had the features of diabetes, one indicator of qi deficiency syndrome, and one or more specialized spleen-related indicators. Conclusion These results suggest that type 2diabetic gastric mucosal injury rat model can be successfully induced by the injection of small-dose streptozotocin( STZ) combined with feeding of high fat and sugar forage and gastric gavage of 30% ethanol solution, and the time window of spleen deficiency for type 2 diabetic gastric mucosal injury rat model is 10-16 weeks after the modeling.
Objective To standardize the modeling and evaluation of type 2 diabetic gastric mucosal injury rat model of spleen deficiency. Methods Male SD rats were randomly divided into normal group and model group,type 2 diabetic rat model with gastric mucosal injury was established by the injection of small-dose streptozotocin(STZ) combined with feeding of high fat and sugar forage and gastric gavage of 30% ethanol solution. After the modeling, we examined the levels of fasting blood glucose( FBG), fasting insulin( FINS), superoxide dismutase(SOD), malondialdehyde(MDA) and nitric oxide(NO) in the rat serum, and gastric and pancreatic tissue pathology was evaluated after HE staining. Modern medical evaluation method was used for the statistics of information collected by four diagnostic methods of traditional Chinese medicine( TCM), and for the identification of properties and evolution of TCM syndromes. Results Type 2 diabetic gastric mucosal injury rat model had been established in 75% of the rats. And 10-16 weeks after the modeling, the model rats had the syndrome of spleen deficiency, showing as persistent emaciation, decreased grasping force, weak activities in open field test, pale tongue nature, decreased r value of the claw and tail, increased defecation frequency,softening of loose stool. Of all the model rats, 78% had the features of diabetes, one indicator of qi deficiency syndrome, and one or more specialized spleen-related indicators. Conclusion These results suggest that type 2diabetic gastric mucosal injury rat model can be successfully induced by the injection of small-dose streptozotocin( STZ) combined with feeding of high fat and sugar forage and gastric gavage of 30% ethanol solution, and the time window of spleen deficiency for type 2 diabetic gastric mucosal injury rat model is 10-16 weeks after the modeling.
引文
[1]孙国鹏,尹国安,张艳芳.糖尿病动物模型研究进展[J].中外医学研究,2013,11(14):149.
[2]田婷,马向华,沈捷.2型糖尿病动物模型研究概况[J].医学综述,2011,17(6):905.
[3]Yang W,Lu J,Weng J,et al.Prevalence of diabetes among men and women in China[J].N Engl J Med,2010,362(12):1090.
[4]郑悦,王化虹,陈宝雯.胃肠道症状与糖尿病关系的分析[J].中国医药导刊,2003(2):82.
[5]Hep A,Dolina J,Prasek J,et al.Dyspeptic complaints in diabetic patients[J].Vnitr Lek,2000,46(11):782.
[6]Hung C R.Effect of lysozyme chloride on betel quid chewing aggravated gastric oxidative stress and hemorrhagic ulcer in diabetic rats[J].World J Gastroenterol,2005,11(37):5853.
[7]Bhor V M,Raghuram N,Sivakami S.Oxidative damage and altered antioxidant enzyme activities in the small intestine of streptozotocin-induced diabetic rats[J].Int J Biochem Cell Biol,2004,36(1):89.
[8]张毅,李慧颖,董玲,等.链脲佐菌素诱导模拟2型糖尿病动物模型的研究进展[J].中华糖尿病杂志,2011,3(5):433.
[9]张永斌,刘晓秋,陈嘉,等.补中益气丸对糖尿病大鼠NO、PGs、MAN1A1相关胃黏膜损伤的修复[J].中国兽医杂志,2013,49(5):51.
[10]黄江荣,杜亚明,鄢进,等.加味苓桂术甘汤对代谢综合征大鼠血清抵抗素、脂联素、胰岛素、胰岛素抵抗的影响[J].中国实验方剂学杂志,2013,19(6):227.
[11]李光伟,潘孝仁,Stephen L,等.检测人群胰岛素敏感性的一项新指数[J].中华内科杂志,1993,32(10):656.
[12]段丽菊,甘耀坤,杨旭,等.两种红细胞超氧化物歧化酶测活方法的比较[J].湖北预防医学杂志,2004,15(1):16.
[13]刘振宗,钱镜秋.红细胞丙二醛硫代巴比妥酸比色法及其初步临床应用[J].陕西医学检验,1993,8(1):1.
[14]郭庆军,段南宁,陈积梅.两种一氧化氮检测试剂盒的比较(摘要)[J].沈阳部队医药,2001,14(5):438.
[15]汤家铭,陈民利.医学实验动物学[M].北京:中国中医药出版社,2012:182,187,198.
[16]潘志强,方肇勤,阚卫兵,等.正常昆明种小鼠后爪图谱及RGB分析[J].上海中医药大学学报,2003,17(3):46.
[17]中华人民共和国卫生部药政局.中药治疗脾虚证的临床研究指导原则[J].中国医药学报,1988,5(3):71.
[18]Kassander P.Asymptomatic gastric retention in diabetics(gastroparesis diabeticorum)[J].Annals of Internal Medicine,1958,48(4):797.
[19]Hung C R,Huang E Y.Role of acid back-diffusion in the formation of mucosal ulceration and its treatment with drugs in diabetic rats[J].Journal of Pharmacy and Pharmacology,1995,47(6):493.
[20]Kong M F,Horowitz M.Gastric emptying in diabetes mellitus:relationship to blood-glucose control[J].Clinics in Geriatric Medicine,1999,15(2):321.
[21]徐文娟,张良晓,黄宇虹,等.基于血浆中脂肪酸代谢谱及非线性判别分析方法的糖尿病中医证候分型[J].色谱,2012(9):864.
[22]黎元元,郭蕾,高思华.基于专家观点的糖尿病证候要素提取文献分析[J].现代中西医结合杂志,2012(6):571.
[23]王洪武,王玉兴.2型糖尿病证候研究规律的思考[J].天津中医药大学学报,2011(1):6.
[24]马会霞,严美花,张浩军,等.糖尿病病证结合动物模型的现状思考[J].中华中医药杂志,2013,28(7):1933.
[25]吴文莉,马威,薛莎,等.肾阴虚型糖尿病动物模型的研制[J].湖北中医学院学报,1999,1(3):27.
[26]吴忆,李敬林,张锐,等.2型糖尿病气阴两虚证动物模型研究及反证中药对证候的影响[J].辽宁中医杂志,2005,32(7):735.
[27]董天宝,李敬林.阴虚热盛型2型糖尿病动物模型的初步研究[J].实用中医内科杂志,2009,23(6):31.
[28]方肇勤.辨证论治研发的一些基本学术问题[J].中国中西医结合杂志,2008,28(7):652.
[29]胡梅芳.糖尿病从脾论治分析[J].安徽中医临床杂志,2003,15(4):351.
[30]孙彪.从脾论治糖尿病[J].中医研究,2001,14(4):12.
[2]田婷,马向华,沈捷.2型糖尿病动物模型研究概况[J].医学综述,2011,17(6):905.
[3]Yang W,Lu J,Weng J,et al.Prevalence of diabetes among men and women in China[J].N Engl J Med,2010,362(12):1090.
[4]郑悦,王化虹,陈宝雯.胃肠道症状与糖尿病关系的分析[J].中国医药导刊,2003(2):82.
[5]Hep A,Dolina J,Prasek J,et al.Dyspeptic complaints in diabetic patients[J].Vnitr Lek,2000,46(11):782.
[6]Hung C R.Effect of lysozyme chloride on betel quid chewing aggravated gastric oxidative stress and hemorrhagic ulcer in diabetic rats[J].World J Gastroenterol,2005,11(37):5853.
[7]Bhor V M,Raghuram N,Sivakami S.Oxidative damage and altered antioxidant enzyme activities in the small intestine of streptozotocin-induced diabetic rats[J].Int J Biochem Cell Biol,2004,36(1):89.
[8]张毅,李慧颖,董玲,等.链脲佐菌素诱导模拟2型糖尿病动物模型的研究进展[J].中华糖尿病杂志,2011,3(5):433.
[9]张永斌,刘晓秋,陈嘉,等.补中益气丸对糖尿病大鼠NO、PGs、MAN1A1相关胃黏膜损伤的修复[J].中国兽医杂志,2013,49(5):51.
[10]黄江荣,杜亚明,鄢进,等.加味苓桂术甘汤对代谢综合征大鼠血清抵抗素、脂联素、胰岛素、胰岛素抵抗的影响[J].中国实验方剂学杂志,2013,19(6):227.
[11]李光伟,潘孝仁,Stephen L,等.检测人群胰岛素敏感性的一项新指数[J].中华内科杂志,1993,32(10):656.
[12]段丽菊,甘耀坤,杨旭,等.两种红细胞超氧化物歧化酶测活方法的比较[J].湖北预防医学杂志,2004,15(1):16.
[13]刘振宗,钱镜秋.红细胞丙二醛硫代巴比妥酸比色法及其初步临床应用[J].陕西医学检验,1993,8(1):1.
[14]郭庆军,段南宁,陈积梅.两种一氧化氮检测试剂盒的比较(摘要)[J].沈阳部队医药,2001,14(5):438.
[15]汤家铭,陈民利.医学实验动物学[M].北京:中国中医药出版社,2012:182,187,198.
[16]潘志强,方肇勤,阚卫兵,等.正常昆明种小鼠后爪图谱及RGB分析[J].上海中医药大学学报,2003,17(3):46.
[17]中华人民共和国卫生部药政局.中药治疗脾虚证的临床研究指导原则[J].中国医药学报,1988,5(3):71.
[18]Kassander P.Asymptomatic gastric retention in diabetics(gastroparesis diabeticorum)[J].Annals of Internal Medicine,1958,48(4):797.
[19]Hung C R,Huang E Y.Role of acid back-diffusion in the formation of mucosal ulceration and its treatment with drugs in diabetic rats[J].Journal of Pharmacy and Pharmacology,1995,47(6):493.
[20]Kong M F,Horowitz M.Gastric emptying in diabetes mellitus:relationship to blood-glucose control[J].Clinics in Geriatric Medicine,1999,15(2):321.
[21]徐文娟,张良晓,黄宇虹,等.基于血浆中脂肪酸代谢谱及非线性判别分析方法的糖尿病中医证候分型[J].色谱,2012(9):864.
[22]黎元元,郭蕾,高思华.基于专家观点的糖尿病证候要素提取文献分析[J].现代中西医结合杂志,2012(6):571.
[23]王洪武,王玉兴.2型糖尿病证候研究规律的思考[J].天津中医药大学学报,2011(1):6.
[24]马会霞,严美花,张浩军,等.糖尿病病证结合动物模型的现状思考[J].中华中医药杂志,2013,28(7):1933.
[25]吴文莉,马威,薛莎,等.肾阴虚型糖尿病动物模型的研制[J].湖北中医学院学报,1999,1(3):27.
[26]吴忆,李敬林,张锐,等.2型糖尿病气阴两虚证动物模型研究及反证中药对证候的影响[J].辽宁中医杂志,2005,32(7):735.
[27]董天宝,李敬林.阴虚热盛型2型糖尿病动物模型的初步研究[J].实用中医内科杂志,2009,23(6):31.
[28]方肇勤.辨证论治研发的一些基本学术问题[J].中国中西医结合杂志,2008,28(7):652.
[29]胡梅芳.糖尿病从脾论治分析[J].安徽中医临床杂志,2003,15(4):351.
[30]孙彪.从脾论治糖尿病[J].中医研究,2001,14(4):12.