滋阴补肾序贯法对多囊卵巢综合征大鼠卵巢StAR蛋白及1基因表达的影响

Effect of Ziyin Bushen sequential method on acute regulatory proteins of steroid hormone and SIRT1 gene expression in ovaries of polycystic ovary syndrome rats

目的探讨滋阴补肾序贯法对多囊卵巢综合征(PCOS)大鼠卵巢类固醇激素合成急性调控蛋白(StAR)及SIRT1基因表达的影响。方法用脱氢表雄酮(DHEA)创建PCOS大鼠模型。80只21日龄清洁级SD大鼠随机分为空白对照组(16只)和PCOS组(64只)。将造模成功大鼠随机分为生理盐水组、滋阴组、补肾组和序贯组,每组各16只。各组连续灌胃20 d。用酶联免疫吸附法(ELISA)测定血清雌二醇、孕酮和睾酮浓度。分别用Western blotting和RT-PCR测定各组卵巢StAR、SIRT1蛋白及mRNA表达。结果生理盐水组血清雌二醇水平较空白对照组显著升高(P=0.004);滋阴组、补肾组和序贯组血清雌二醇水平较生理盐水组显著降低(P=0.005、P=0.003、P=0.002)。生理盐水组血清睾酮水平显著高于空白对照组(P=0.005);序贯组血清睾酮水平较生理盐水组、滋阴组和补肾组均显著降低(P=0.002、P=0.005和P=0.005)。与空白对照组相比,生理盐水组StAR蛋白含量高(P<0.001),而SIRT1蛋白显著减少(P<0.001);序贯组StAR蛋白及mRNA水平较生理盐水组、滋阴组和补肾组均显著降低(P<0.001、P=0.008、P<0.001);序贯组SIRT1蛋白表达水平较生理盐水组和补肾组均显著升高(P均<0.001);与空白对照组相比,生理盐水组StAR mRNA显著升高(P<0.001),而SIRT1 mRNA显著减少(P<0.001);滋阴组和序贯组SIRT1 mRNA水平较生理盐水组均有显著升高(P均<0.001),而序贯组比补肾组显著升高(P<0.001)。结论滋阴补肾序贯法可能通过SIRT1基因高表达调控卵巢局部StAR等类固醇激素合成相关蛋白,而降低PCOS大鼠血清高雄激素血症。

Objective To investigate the effect of Ziyin Bushen sequential method on the expression of steroids acute regulatory synthesis protein(StAR)and silent information regulator 1(SIRT1)in ovaries of polycystic ovary syndrome(PCOS)rats.Methods PCOS rat model was established by dehydroepiandrosterone(DHEA).Eighty 21-day-old clean SD rats were randomly divided into control group(16 rats)and PCOS group(64 rats).PCOS group was further divided into normal saline(NS)group,Ziyin group,Bushen group and Xuguan group with 16 rats in each group.Each group was given intragastric administration for 20 d.Serum estradiol,progesterone and testosterone levels were measured by enzyme linked immunosorbent assay(ELISA)method.The expression levels of StAR,SIRT1 protein and mRNA were detected by Western blotting and RT-PCR.Results The serum estradiol level in normal saline group was significantly higher than that in control group(P=0.004).The levels of serum estradiol in Ziyin group,Bushen group and Xuguan group were significantly lower than that in NS group(P=0.005,P=0.003 and P=0.002,respectively).The serum testosterone level in normal saline group was significantly higher than that in control group(P=0.005);the level of serum testosterone in Xuguan group was significantly lower than that in NS group,Ziyin group and Bushen group(P=0.002,P=0.005 and P=0.005,respectively).Compared with control group,the StAR protein content in the normal saline group was higher(P<0.001),while SIRT1 mRNA was significantly decreased(P<0.001).Compared with NS group,Ziyin group and Bushen group,the expression of StAR protein and mRNA in Xuguan group was significantly lower(P<0.001,P=0.008 and P<0.001,respectively);the expression of SIRT1 protein in Xuguan group was significantly higher than that in NS group and Bushen group(all P<0.001);compared with the control group,StAR mRNA in NS group was significantly increased(P<0.001),while SIRT1 mRNA was significantly decreased(P<0.001);the expression of SIRT1 gene in Ziyin group and Xuguan group was significantly higher than that in NS group(all P<0.001),while that in Xuguan group was significantly higher than that in Bushen group(P<0.001).Conclusion Ziyin Bushen sequential method may decrease the serum androgen level of PCOS rats by upregulating SIRT1 gene expression and StAR in ovary expression.