左归丸对化疗性卵巢早衰小鼠卵巢组织中缝隙连接蛋白43表达的影响

Effect of Zuogui Pill on the Expression of Gap Junction Protein 43 in Ovarian Tissue of Mice with Chemotherapy-induced Premature Ovarian Failure

目的:研究左归丸对环磷酰胺(CTX)致卵巢早衰(POF)小鼠卵巢组织中缝隙连接蛋白43(Cx43)表达的影响,探讨左归丸治疗化疗性POF的作用机制。方法:将72只小鼠按体质量随机分为空白组、模型组、戊酸雌二醇片组(阳性对照,0.000 13 g/kg)和左归丸低、中、高剂量组(13.65、40.95、122.85 g/kg),每组10只。除空白组外,其余各组小鼠均ip CTX复制POF模型,每天1次,连续20 d;并同时ig相应药物,每天1次,连续30 d。末次给药2h后,分别采用反转录-聚合酶链反应法和Western blot法检测卵巢组织中Cx43 mRNA和蛋白表达;并采用免疫组化法检测卵巢组织中Cx43蛋白的分布。结果:与空白组比较,模型组大鼠卵巢组织中Cx43 mRNA及蛋白表达均明显减弱,卵泡与颗粒细胞中Cx43蛋白的分布明显减少(P<0.01);与模型组比较,戊酸雌二醇片组和左归丸中、高剂量组小鼠卵巢组织中Cx43 mRNA及蛋白表达均明显增强,卵泡与颗粒细胞中Cx43蛋白的分布明显增加(P<0.01)。结论:左归丸可上调CTX致POF小鼠卵巢组织中Cx43 mRNA及蛋白表达,增加Cx43在卵泡及颗粒细胞间的分布,改善缝隙连接功能,这可能是其治疗POF的作用机制之一。

OBJECTIVE：To study the effect of Zuogui pill on the expression of gap junction protein 43（Cx43）in ovarian tissue of mice with premature ovarian failure（POF）induced by intraperitoneal injection of cyclophosphamide（CTX）,and explore its mechanism in the treatment of chemotherapy-induced POF. METHODS：72 mice were randomly divided into blank group,model group,Estradiol valerate tablet group（positive control,0.000 13 g/kg）,Zuogui pill low-dose,medium-dose,high-dose groups（13.65,40.95,122.85 g/kg）by body mass,10 in each group. Except for blank group,other groups were reduce POF model by ip CTX,once a day,for 20 d. Meanwhile,the mice were intragastrically administrated related medicines,once a day,for 30 d. After2 h of last administration,reverse transcription-polymerase chain reaction method and Western blot method were used to detect the Cx43 mRNA and protein expressions in ovarian tissue respectively,and immunohistochemistry method was adopted to detect the distribution of Cx43 protein in ovarian tissue. RESULTS：Compared with blank group,Cx43 mRNA and protein expressions in ovarian tissue of mice in model group were obviously weakened,and the distribution of Cx43 protein in follicle and granulocytes were obviously reduced（P〈0.01）. Compared with model group,Cx43 mRNA and protein expressions in ovarian tissue of mice in Estradiol valerate tablet group and Zuogui pill medium-dose,high-dose groups were obviously strengthened,and the distribution of Cx43 protein in follicle and granulocytes were obviously increased（P〈0.01）. CONCLUSIONS：Zuogui pill can increase the Cx43 mRNA and protein expressions in ovarian tissue of CTX-induced POF mice,increase the distribution of Cx43 in follicle and granulocytes and gap junction function,which may be one of the treatment mechanism of POF.