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目的:探讨何首乌饮对衰老大鼠胰岛素信号通路的影响。选用30只雄性大鼠随机分为3组:青年组(12月龄);衰老组(18月龄);何首乌饮组(何首乌饮4.8g/100g,灌胃60d,18月龄),每组各10只。采用β-半乳糖苷酶染色观察何首乌饮延缓睾丸衰老的情况,基因芯片技术筛选衰老大鼠受何首乌饮调控的差异表达基因群体,并采用qRT-PCR进行验证。从中筛选出受何首乌饮调控与胰岛素信号通路相关的关键基因IR、IRS1、IRS2、IGF1、IGFBP3,进行qRT-PCR、免疫荧光和Western blot观察其在睾丸组织中的表达变化。结果:衰老组大鼠睾丸组织衰老细胞明显高于青年组(P<0.01),何首乌饮用药后衰老细胞明显减少(P<0.01)。基因芯片筛选出受何首乌饮直接调控的基因有912个,其中与胰岛素信号通路相关的基因有10个,从中选取的关键基因,衰老组IGFBP3的表达高于青年组(P<0.01),IR、IRS1、IRS2、IGF1表达低于青年组(P<0.01),用药后IGFBP3的表达下调,IR、IRS1、IRS2、IGF1表达上调(P<0.01)。结论:何首乌饮通过调节胰岛素信号通路关键基因的表达延缓大鼠睾丸衰老。
Objective: To investigate the effect of Heshouwuyin on insulin signaling pathway in aging rats. A total of 30 male rats were randomly divided into 3 groups: young group (12 months), aging group (18 months old), Polygonum multiflorum group (4.8g / 100g, gavage 60d, 18 months) Each 10. The β-galactosidase staining was used to observe the decoction of Polygonum multiflorum Thunb. To delay the testicular senescence. The cDNA microarray was used to screen the differentially expressed genes controlled by the Polygonum multiflorum Thunb. In the aged rats by gene chip technology and validated by qRT-PCR. The key genes IR, IRS1, IRS2, IGF1 and IGFBP3, which are related to the insulin signaling pathway, were screened out by qingwu decoction. QRT-PCR, immunofluorescence and Western blot were used to detect the expression of IR, IRS1, IRS2 and IGFBP3 in testis. Results: The senescent cells of testis in aged rats were significantly higher than those in young rats (P <0.01). There were 912 genes directly screened out by He Shouyin, including 10 genes related to insulin signaling pathway. The expression of IGFBP3 in aging group was higher than that in young group (P <0.01) The expression of IRS1, IRS2 and IGF1 was lower than that of the young group (P <0.01). The expression of IGFBP3 was down-regulated and the expressions of IR, IRS1, IRS2 and IGF1 were up-regulated after treatment (P <0.01). Conclusion: Heshouwuyin can delay the testicular aging in rats by regulating the expression of key genes of insulin signaling pathway.