基于线粒体通路探讨右归颗粒改善肾阳虚IVF患者治疗结局的机制研究

发布时间：2018-04-12 19:55

本文选题：右归颗粒 + 肾主生殖　；参考：《山东中医药大学》2016年硕士论文

【摘要】：目的:探讨右归颗粒对肾阳虚型不孕症患者卵巢颗粒细胞线粒体凋亡通路关键效应分子Bax、Cyt-c、Caspase-3的干预作用及其对卵细胞质量、妊娠结局的影响,探究肾阳虚致不孕在线粒体凋亡通路水平可能的发生机制及补肾中药在此水平可能的作用机制。方法:选取因输卵管因素行体外受精-胚胎移植(IVF-ET)术治疗的肾阳虚不孕症患者70例,根据患者就诊次序,随机分为右归颗粒治疗组与安慰剂组各35例,另选取因男方因素行IVF的正常对照组患者35例。治疗组于卵泡期始服用右归颗粒联合常规IVF治疗,安慰剂组及正常对照组患者同一时期服用安慰剂联合常规IVF治疗。观察肾阳虚患者治疗前后肾阳虚症候积分变化;三组患者HCG日血E2、LH、P水平;优质卵率、优质胚胎率、临床妊娠率以及取卵日卵巢颗粒细胞线粒体通路中Bax、Cyt-c、Caspase-3在基因及蛋白水平的表达。结果:1、右归颗粒组患者治疗后肾阳虚症状明显改善,与安慰剂组相比,优卵率、优胚率、临床妊娠率均增高明显(P0.05);2.比较三组患者卵巢颗粒细胞线粒体通路中关键效应分子Bax及其下游Cyt-c、Caspase-3基因及蛋白表达,发现:治疗组与安慰剂组相比,BaxmRNA表达下调,全细胞水平Cyt-c、Caspase-3蛋白含量降低,差异有统计学意义(P0.05);安慰剂组与正常对照组相比,BaxmRNA表达上调,全细胞水平Cyt-c、Caspase-3蛋白含量明显升高,差异有统计学意义(P0.05)。结论:1、右归颗粒可明显改善肾阳虚型不孕症患者的肾虚状态,提高其优卵率、优胚率,改善临床妊娠率;2、右归颗粒可能是通过抑制线粒体凋亡通路,下调Bax因子、抑制Cty-c自线粒体中释放,控制Caspase-3表达,进而抑制颗粒细胞凋亡,改善肾阳虚不孕症患者的IVF结局;3、从线粒体凋亡通路角度阐释了“补肾促生殖”的作用机制,为“肾主生殖”理论提供了较为客观的依据。
[Abstract]:Objective: to investigate the intervention effect of Yougui granule on mitochondrial apoptosis pathway of ovarian granulosa cells in patients with kidney-yang deficiency type infertility and its effect on the quality of oocyte and pregnancy outcome.To explore the possible mechanism of apoptosis pathway in mitochondria caused by deficiency of kidney-yang and the possible action mechanism of traditional Chinese medicine for tonifying kidney.Methods: seventy patients with kidney yang deficiency infertility treated by in vitro fertilization and embryo transfer with IVF-ETS due to fallopian tube factors were randomly divided into two groups: the right Gui granule group (35 cases) and the placebo group (35 cases).Another 35 cases of normal control group who were treated with IVF because of male factors were selected.The patients in the treatment group were treated with the right Gui granule combined with conventional IVF at the follicular stage, and the patients in the placebo group and the normal control group were treated with placebo combined with conventional IVF at the same time.To observe the changes of the symptoms of kidney yang deficiency before and after treatment, to observe the level of E2LHU P in HCG daily blood, to observe the rate of high quality eggs, and to observe the rate of high quality embryos in the three groups.Clinical pregnancy rate and the expression of Bax-Cyt-Caspase-3 in the mitochondrial pathway of ovarian granulosa cells on the day of egg taking.Results the symptoms of deficiency of kidney yang were obviously improved after treatment in the control group. Compared with the placebo group, the excellent egg rate, the optimal embryo rate and the clinical pregnancy rate were significantly higher than those in the placebo group (P 0.05).The expression of key effector molecule Bax and its downstream Cyt-caspase-3 gene and protein in the mitochondrial pathway of ovarian granulosa cells were compared among the three groups. The results showed that the expression of BaxmRNA was down-regulated and the content of Cyt-caspase-3 was decreased at the whole cell level in the treatment group compared with the placebo group.Compared with the control group, the expression of BaxmRNA in placebo group was up-regulated, and the protein content of Cyt-caspase-3 in the whole cell level was significantly increased (P 0.05).Conclusion: W1, Yougui granule can obviously improve the state of kidney deficiency, increase the rate of excellent egg and embryo, and improve the clinical pregnancy rate of infertile patients with deficiency of kidney-yang. Yougui granule may be by inhibiting mitochondrial apoptosis pathway, down-regulating Bax factor.Inhibiting the release of Cty-c from mitochondria, controlling the expression of Caspase-3, inhibiting the apoptosis of granulosa cells and improving the outcome of IVF in patients with infertility due to deficiency of kidney yang, the mechanism of "tonifying the kidney and promoting reproduction" was explained from the perspective of mitochondrial apoptosis pathway.It provides an objective basis for the theory of "kidney dominating reproduction".
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R271.14

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