人参-三七-川芎提取物延缓高糖诱导的小鼠血管衰老的机制探讨

目的:从单磷酸腺苷活化蛋白激酶/哺乳动物雷帕霉素靶蛋白(AMPK/mTOR)通路探讨人参-三七-川芎提取物对高糖诱导的小鼠血管衰老的保护作用。方法:130只雄性C57BL/6小鼠先随机分为空白组和高糖组,高糖组腹腔注射链脲佐菌素(STZ),并用高脂饮食连续性喂养7个月后再次随机分组,分为模型组、人参-三七-川芎提取物低剂量组(0.819 g·kg~(-1))、人参-三七-川芎提取物高剂量组(1.638 g·kg~(-1))及二甲双胍组(150 mg·kg~(-1)),灌胃给药,每天1次,连续9周,期间检测小鼠体质量和血糖变化。给药结束后,苏木素-伊红(HE)染色检测小鼠胸主动脉形态,蛋白免疫印迹法(Western blot)检测小鼠胸主动脉周期蛋白依赖激酶抑制因子2A(p16),周期依赖性蛋白激酶抑制因子1A(p21),AMPK,p-AMPK,mTOR,pmTOR,肝激酶B1(LKB1),p-LKB1,核糖体蛋白s6激酶(p70s6k),p-p70s6k蛋白表达情况。结果:与空白组相比,模型组小鼠体质量显著降低、血糖显著升高(P0.01),在药物干预9周后,各给药组小鼠体质量无明显差异,血糖值较模型组明显下降(P0.05,P0.01)。与空白组相比,模型组内膜损伤严重且有增生,中膜明显增生且排列不整齐,给药各组内膜损伤不明显,中膜少量增生,p16,p21,mTOR,p-mTOR,p70s6k,p-p70s6k蛋白表达明显升高(P0.05,P0.01),AMPK,p-AMPK,LKB1,p-LKB1蛋白表达明显下降(P0.05,P0.01)。药物干预后,各给药组p16,p21,mTOR,p-mTOR,p70s6k,p-p70s6k蛋白表达明显下降(P0.05,P0.01),AMPK,p-AMPK,LKB1,p-LKB1蛋白表达明显升高(P0.05,P0.01)。结论:高糖能够诱导小鼠主动脉衰老,人参-三七-川芎提取物可能通过AMPK/mTOR通路改善由高糖诱导的小鼠主动脉衰老。

Extracts from Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma in Delaying High Glucose-induced Vascular Senescence in Mice

Objective: To investigate the protective effect of extracts from Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on vascular senescence induced by high glucose in mice from adenosine 5''-monophosphate (AMP)-activated protein kinase/mechanistic target of rapamycin (AMPK/mTOR) pathway. Method: A total of 130 male C57BL/6 mice were randomly divided into control group and high glucose group. The high glucose group was intraperitoneally injected with streptozocin(STZ) and fed with a high-fat diet continuously for seven months. Mice were divided into 4 groups:model group, low-dose extracts from Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma(0.819 g·kg^-1) group, high-dose extracts from Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma group (1.638 g·kg^-1) and metformin group (150 mg·kg^-1), and intragastrically administered once a day for nine weeks. The changes in body weight and blood glucose were measured. At the end of the administration, htoxylin eosin(HE) was performed for the detection of aortic morphology, and the expressions of cyclin-dependent kinase inhibitor 2A (p16), cyclin-dependent kinase inhibitor 1A (p21), AMPK, p-AMPK, mTOR, p-mTOR, liver kinase B1 (LKB1), p-LKB1, Ribosomal protein s6 kinase (p70s6k) and p-p70s6k proteins in mouse aorta were detected by Western blot. Result: Compared with blank group, mice in model group had lower body weight and higher blood glucose (P<0.01). After 9 weeks of drug intervention, there was no significant difference in body weight among groups, and the blood glucose level was significantly lower than that in model group (P<0.05, P<0.01). Model group showed a severe intima injury and hyperplasia, middle membrane was obviously proliferated and irregularly arranged. After drug intervention, the intimal damage of each group was not obvious, and the middle membrane was slightly proliferated. The protein expressions of p16, p21, mTOR, p-mTOR, p70s6k and p-p70s6k in model group were significantly higher than those in control group (P<0.05, P<0.01), and the protein expressions of AMPK, p-AMPK, LKB1 and p-LKB1 were significantly decreased (P<0.05, P<0.01). After drug intervention, the protein expressions of p16, p21, mTOR, p-mTOR, p70s6k, p-p70s6k in each group were significantly decreased (P<0.05, P<0.01), while the protein expressions of AMPK, p-AMPK, LKB1, p-LKB1 were significantly increased (P<0.05, P<0.01). Conclusion: High glucose can induce vascular senescence, and extracts from Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma can improve vascular aging induced by high glucose through AMPK/mTOR pathway.