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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(P<0.05);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(P<0.05)。与空白组比较,模型组NFAT-4的蛋白含量明显升高(P<0.05),而miR-133a表达水平明显降低(P<0.05);与模型组比较,各治疗组miR-133a表达水平明显明显升高(P<0.05或P<0.01),而NFAT-4的蛋白含量明显降低(P<0.05或P<0.01)。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。
Objective To investigate the myocardial protective effect of Yiqi Yangyin Huoxue Recipe on diabetic rats. Methods Sixty Wistar male rats, aged 12 weeks, were randomly divided into blank group (n = 10) and model group (n = 50). After the model was successfully established, 50 diabetic rats were randomly divided into 5 groups: normal saline, compound Danshen dripping pills 70 mg / kg). The high, middle and low dose (55.00,27.50,13.75 g / kg) of Yiqi Yangyin Huoxue was given into the stomach once a day for 4 weeks. Blood samples were taken for determination of blood glucose, blood lipid and other biochemical parameters , And the left ventricular anterior myocardial tissue was taken from the rats. One part was made into sections and stained with hematoxylin and eosin (HE), and the morphological changes of myocardial cells were observed by light microscopy. The other part was assayed by Western blot to detect NFAT-4, Quantitative PCR (qRT-PCR) was used to detect the expression of miR-133a. Results Compared with the model group, the levels of blood glucose, glycosylated serum protein and lipids in the model group and the blank group were significantly increased (P <0.05) P <0.05). Compared with the blank group, the protein level of NFAT-4 in the model group was significantly increased (P <0.05), while the expression level of miR-133a was significantly decreased (P <0.05); Compared with the model group, the expression level of miR-133a in each treatment group was significantly (P <0.05 or P <0.01), while the protein content of NFAT-4 decreased significantly (P <0.05 or P <0.01). Yiqi Yangyin Huoxue Recipe and Compound Danshen Dripping Pills can improve the pathological morphology of cardiomyocytes under light microscope. Conclusion Yiqi Yangyin Huoxue can significantly improve glucose and lipid metabolism and inhibit cardiac hypertrophy in diabetic rats and its protective effect on cardiomyocytes may be related to the regulation of the expression of miR-133a and the inhibition of cardiac hypertrophy related protein NFAT-4 .