基于SIRT1/P53通路探讨补肾解郁调冲方改善PCOS合并慢性心理应激大鼠卵巢/海马功能的作用机制

Mechanism of Bushen Jieyu Tiaochong Formula on improving ovarian/hippocampal function in rats with PCOS combined with CUMS based on SIRT1/P53 signaling pathway

目的:观察补肾解郁调冲方对多囊卵巢综合征(PCOS)合并慢性不可预知温和刺激(CUMS)大鼠卵巢/海马功能的调节作用及对SIRT1/P53通路的调控作用。方法:将40只具有正常动情周期的大鼠随机分为空白组、模型组、中药组及西药组,除空白组外均采用来曲唑联合CUMS法造模28 d,造模完成后,中药组予补肾解郁调冲方治疗,西药组予达英-35治疗。观察指标包括卵巢组织形态学、血清性激素水平等PCOS病理状态的变化,旷场试验、海马组织形态学、血清神经递质水平等慢性应激病理状态的变化,以及卵巢及海马组织内SIRT1/P53信号通路蛋白的表达情况。结果:与空白组比较,模型组大鼠卵巢呈多囊样改变,促黄体生成素(LH)、游离睾酮(FT)显著升高(P<0.01);旷场试验活动次数减少(P<0.05),海马神经元细胞形态改变,血清五羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)水平降低(P<0.01,P<0.05)。与模型组比较,各治疗组PCOS病理状态及慢性应激病理状态均不同程度的改善。模型组卵巢及海马组织SIRT1显著降低(P<0.05),卵巢组织乙酰化P53(Acetyl-P53)水平显著升高(P<0.05);中药组卵巢及海马组织SIRT1显著升高(P<0.05),卵巢组织Acetyl-P53水平显著降低(P<0.05)。结论:补肾解郁调冲方能够改善PCOS合并CUMS大鼠卵巢/海马功能,其作用机制可能与SIRT1/P53信号通路调节卵巢及海马细胞凋亡相关。

Objective:To observe the regulatory effect of Bushen Jieyu Tiaochong Formula on ovarian/hippocampal function in rats with PCOS combined with chronic unpredictable mild stress(CUMS),and to investigate its regulatory effects on SIRT1/P53 signaling pathway.Methods:Forty rats with normal motility cycles were screened and divided randomly into blank group,model group,Chinese medicine group and western medicine group.All groups except the blank group were treated with letrozole combined with CUMS for 28 d.After the completion of modeling the Chinese medicine group was treated with the Bushen Jieyu Tiaochong Formula,and the Western medicine group was treated with Diane-35.The observation indicators included changes in the pathological status of PCOS such as ovarian histomorphology and serum sex hormone levels,as well as changes in the pathological status of chronic stress such as absentee field test,hippocampal histomorphology and serum neurotransmitter levels,respectively,and the expression of SIRT1/P53 signaling pathway proteins in ovarian and hippocampal tissues.Results:Compared with the blank group,rats in the model group showed polycystic-like changes in ovaries,significantly higher LH and FT(P<0.01);reduced activity in the absentee test(P<0.05),altered hippocampal neuronal cell morphology and reduced serum 5-HT,DA and NE levels(P<0.01,P<0.05).Compared with the model group,all treatment groups showed different degrees of improvement in PCOS pathology and chronic stress pathology.In the model group,SIRT1 in ovarian and hippocampal tissues was significantly reduced(P<0.05),and acetylated P53(Acetyl-P53)levels in ovarian tissues were significantly increased(P<0.05);in the Chinese medicine group,SIRT1 in ovarian and hippocampal tissues was significantly increased(P<0.05)and Acetyl-P53 levels in ovarian tissues were significantly reduced(P<0.05).Conclusion:Bushen Jieyu Tiaochong Formula can improve ovarian/hippocampal function in PCOS combined with CUMS rats,and its mechanism of action may be related to the SIRT1/P53 signaling pathway regulating ovarian and hippocampal cell apoptosis.