黄芪甲苷延缓肾脏衰老的作用及机制研究。

Zi-Yuan Zhang, Jing-Ai Fang, Su-Fen Li, Ya-Ling Hu, Wen-Yuan Liu, Xue-Jun Liu
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引用次数: 0

摘要

目的:探讨黄芪甲苷Ⅳ通过调节SIRT1/p53信号通路抑制大鼠细胞凋亡、延缓肾脏衰老的机制。方法:采用皮下注射d -半乳糖200 mg/(kg·d)建立衰老模型。将spf级健康雄性SD大鼠随机分为4组:正常对照组(灌胃5 ml/(kg·d)生理盐水)、衰老模型组(灌胃5 ml/(kg·d)生理盐水)、黄芪甲苷组(灌胃40 mg/(kg·d)黄芪甲苷)、SRT1720组(灌胃20 mg/(kg·d) SRT1720),每组10只。8周后采集大鼠血清,采用ELISA法检测大鼠肾功能(肌酐、尿素氮)水平及衰老相关分泌表型(TGF-β、IL-6)。取大鼠肾组织进行HE和Masson染色。采用Western blot和RT-PCR检测SIRT1、p53、Bcl-2、Bax、p21和pRb蛋白及mRNA的表达。结果:老龄模型组大鼠血清肌酐、尿素氮水平高于正常组,但各组间差异无统计学意义(P>0.05)。衰老模型组大鼠血清TGF-β、IL-6水平高于正常组(P<0.05),黄芪甲苷组、SRT1720组大鼠血清TGF-β、IL-6水平低于模型组(P<0.05)。黄芪甲苷组与SRT1720组比较,差异无统计学意义(P>0.05)。肾组织病理染色结果显示,与正常组比较,衰老模型组大鼠肾小管扩张,局部萎缩,炎症细胞浸润,胶原纤维增生。与衰老模型组比较,黄芪甲苷组和SRT1720组大鼠的病理改变均有所减轻。Western blot和RT-PCR结果显示,与正常组比较,衰老组肾组织中SIRT1、pRb蛋白和mRNA表达量降低,Bcl-2蛋白表达量降低(P<0.05), p53、p21蛋白和mRNA表达量升高,Bax蛋白表达量升高(P<0.05)。与衰老组比较,黄芪甲苷和SRT1720均改善了上述指标(P<0.05)。结论:黄芪甲苷可通过调节SIRT1/p53信号通路延缓肾脏衰老。
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[Effects of astragaloside IV on delaying kidney aging and its mechanisms].

Objective: To investigate the mechanisms of Astragaloside Ⅳ on inhibiting apoptosis and delaying kidney aging in rats by regulating SIRT1/p53 signaling pathway.

Methods: The aging model was established by subcutaneous injection of D-galactose 200 mg/(kg·d). SPF-grade healthy male SD rats were randomly divided into 4 groups: normal control group (intragastric infusion of 5 ml/(kg·d) normal saline), aging model group (intragastric infusion of 5 ml/(kg·d) normal saline), Astragaloside IV group (intragastric infusion of 40 mg/(kg·d) Astragaloside IV),and SRT1720 group( intragastric infusion of 20 mg/(kg·d) SRT1720), with 10 rats in each group. After 8 weeks, the serum samples of rats were collected to detect the levels of renal function (creatinine and urea nitrogen) and senescent associated secretory phenotype (TGF-β and IL-6) by ELISA. The renal tissues of rats were obtained for HE and Masson staining. The protein and mRNA expressions of SIRT1, p53, Bcl-2, Bax, p21 and pRb were detected by Western blot and RT-PCR.

Results: Serum creatinine and urea nitrogen levels in the aging model group were higher than those in the normal group, but there was no significant difference in each group (P>0.05). The serum levels of TGF-β and IL-6 in the aging model group were higher than those in the normal group (P<0.05), and which in the Astragaloside IV group and SRT1720 group were lower than those in the model group (P<0.05). There was no significant differences between Astragaloside IV group and SRT1720 group (P>0.05). The results of pathological staining of renal tissues showed that, compared with the normal group, the renal tubules dilated, local atrophy, infiltration of inflammatory cells and proliferation of collagen fibers were observed in the aging model group. Compared with the aging model group, the pathological changes were alleviated in Astragaloside IV group and SRT1720 group. The results of Western blot and RT-PCR showed that, compared with the normal group, the protein and mRNA expressions of SIRT1 and pRb in the renal tissue of the aging group were decreased, the protein expression of Bcl-2 was decreased(P<0.05), and the protein and mRNA expressions of p53 and p21 were increased, the protein expression of Bax was increased(P<0.05). Compared with the aging group, Astragaloside IV and SRT1720 improved the above-mentioned indexes (P<0.05).

Conclusion: Astragaloside IV can delay kidney aging by regulating the SIRT1/p53 signaling pathway.

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