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益气养阴活血方对2型糖尿病大鼠心肌微小基因miR-133a表达水平的影响
引用本文:张军,刘颖,赵静,刘晓曼,王明星,王军媛,王海英,戴金颖,袁洁,王顺意.益气养阴活血方对2型糖尿病大鼠心肌微小基因miR-133a表达水平的影响[J].北京中医药大学学报,2017,40(3).
作者姓名:张军  刘颖  赵静  刘晓曼  王明星  王军媛  王海英  戴金颖  袁洁  王顺意
作者单位:河北省唐山市中医医院内分泌一科 河北 063000;华北理工大学
基金项目:河北省中医药管理局资助课题
摘    要:目的探讨益气养阴活血方对糖尿病大鼠的心肌保护作用。方法 12周龄Wistar雄性大鼠60只,随机选取空白组10只,造模组50只。采用高脂饮食加小剂量链尿佐菌素(STZ)诱导形成2型糖尿病大鼠模型,造模成功后,50只糖尿病大鼠随机分为5组,分别予生理盐水,复方丹参滴丸(70 mg/kg),益气养阴活血方高、中、低剂量(55.00、27.50、13.75 g/kg)灌胃,每日1次,干预4周,常规取血测定血糖、血脂等生化指标,并取大鼠左室前壁心肌组织,一部分制成切片HE染色后光学显微镜观察心肌细胞形态学改变,另一部分用Western blot法测定活化T细胞核因子4(NFAT-4)含量,运用实时荧光定量PCR(qRT-PCR)技术检测微小基因miR-133a的表达水平。结果模型组与空白组比较大鼠的血糖、糖化血清蛋白、血脂水平均明显升高(P0.05);与模型组比较,各治疗组大鼠血糖、糖化血清蛋白、血脂水平均明显降低(P0.05)。与空白组比较,模型组NFAT-4的蛋白含量明显升高(P0.05),而miR-133a表达水平明显降低(P0.05);与模型组比较,各治疗组miR-133a表达水平明显明显升高(P0.05或P0.01),而NFAT-4的蛋白含量明显降低(P0.05或P0.01)。益气养阴活血方及复方丹参滴丸均可改善心肌细胞光镜下病理形态。结论益气养阴活血方可显著改善糖尿病大鼠糖脂代谢,抑制心肌肥大,其对心肌细胞的保护作用,可能与调控miR-133a的表达、从而抑制心肌肥大相关蛋白NFAT-4的表达有关。

关 键 词:益气养阴活血方  糖尿病心肌病变  miR-133a  糖尿病大鼠

Influence of Yiqi Yangyin Huoxue Fang on expression level of myocardial miR-133a in rats with type 2 diabetes mellitus
ZHANG Jun,LIU Ying,ZHAO Jing,LIU Xiaoman,WANG Mingxing,WANG Junyuan,WANG Haiying,DAI Jinying,YUAN Jie,WANG Shunyi.Influence of Yiqi Yangyin Huoxue Fang on expression level of myocardial miR-133a in rats with type 2 diabetes mellitus[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(3).
Authors:ZHANG Jun  LIU Ying  ZHAO Jing  LIU Xiaoman  WANG Mingxing  WANG Junyuan  WANG Haiying  DAI Jinying  YUAN Jie  WANG Shunyi
Abstract:Objective To investigate the myocardial protective effect of Yiqi Yangyin Huoxue Fang (YYHF) in rats with type 2 diabetes mellitus (T2DM).Methods Male Wistar rats (aged 12 weeks old, n=60) were chosen and divided randomly into blank group (n=10) and modeling group (n=50).The model of T2DM was induced by high-fat diet and low-dose streptozocin (STZ).After successfully modeling, the modeling group were randomly divided into groups and given orally, respectively, normal saline water (model group), Fufang Danshen Diwan(FDD group, 70 mg/kg), and high-dose, mid-dose and low-dose YYHF(high-dose group, mid-dose group, low-dose group, 55.00 g/kg, 27.50 g/kg, 13.75 g/kg).once a day for 4 weeks.The routine biochemical indexes including blood sugar and blood fat were detected.The myocardial tissue was collected from the anterior left ventricular wall, and then a part of which was prepared to slices for observing myocardial cytomorphologic changes after HE stain by using light microscope, and other part of which was taken for detecting content of nuclear factor of activated T cell 4 (NFAT-4) by using Western Blot, and expression of miR-133a by using real-time fluorescence quantitative polymerase chain reaction (RT-PCR).Results The levels of blood sugar, glycated serum protein (GSP) and blood fat increased significantly in model group compared with blank group (P<0.05), and decreased significantly in all treatment groups compared with model group (P<0.05).The content of NFAT-4 increased significantly (P<0.05), and expression of miR-133a decreased significantly (P<0.05) in model group compared with blank group.The expression of miR-133a increased significantly (P<0.05 or P<0.01), and content of NFAT-4 decreased significantly (P<0.05 or P<0.01) in all treatment groups compared with model group.YYHF and FDD alleviated myocardial cytomorphologic changes under light microscope.Conclusion YYHF can significantly ameliorate glycolipid metabolism, inhibit cardiomyocyte hypertrophy and protect myocardial cells, which may be related to the regulation of expressions of miR-133a and NFAT-4.
Keywords:Yiqi Yangyin Huoxue Fang  diabetic cardiomyopathy  miR-133a  rats with diabetes mellitus
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