自拟更年汤对肾阳虚型围绝经期模型大鼠卵巢细胞凋亡相关因子的影响

Effect of Gennian docoction on ovarian cell apoptosis-related factors in the rats with perimenopause of Kidney-Yang deficiency type

目的探讨自拟更年汤对肾阳虚型围绝经期模型大鼠卵巢组织形态学及卵巢颗粒细胞凋亡因子bax、bax mRNA、bcl-2、bcl-2 mRNA的影响。方法选用自然老化的围绝经期雌性SD大鼠,臀部肌肉左右交替肌注氢化可的松构建肾阳虚型围绝经期大鼠模型,将造模成功大鼠随机分为更年汤组、教材方组、利维爱组、生理盐水组;另选择4月龄SD雌性大鼠20只作为青年组,18月龄SD雌性大鼠20只作为老年组。青年组、老年组、生理盐水组均给予生理盐水灌胃,各给药组给予相应药物灌胃,灌胃15 d后处死大鼠,分离摘取双侧卵巢,运用病理切片观察卵巢组织大体形态,运用免疫组化及原位杂交法检测卵巢颗粒细胞凋亡因子bax、bax mRNA、bcl-2、bcl-2 mRNA表达情况。结果各给药组卵泡发育良好情况优于老年组和生理盐水组(P均<0.05),卵泡闭锁、卵泡扩张情况轻于老年组和生理盐水组(P均<0.05);自拟更年汤组与利维爱组卵泡发育良好情况优于教材方组(P均<0.05),自拟更年汤组与利维爱组比较差异无统计学意义(P>0.05);自拟更年汤组卵泡闭锁、卵泡扩张情况轻于教材方组和利维爱组(P均<0.05),教材方组与利维爱组比较差异均无统计学意义(P均>0.05)。各给药组bax、bax mRNA阳性表达明显低于老年组和生理盐水组(P均<0.05),bcl-2与bcl-2 mRNA阳性表达明显高于老年组和生理盐水组(P均<0.05);自拟更年汤组与利维爱组bax、bax mRNA阳性表达明显低于教材方组(P均<0.05),bcl-2与bcl-2 mRNA阳性表达明显高于教材方组(P均<0.05),自拟更年汤组bax、bax mRNA、bcl-2、bcl-2 mRNA阳性表达与利维爱组比较差异无统计学意义(P均>0.05)。结论自拟更年汤能促进卵泡发育,抑制卵泡闭锁和扩张,能上调卵巢颗粒细胞抑制凋亡因子bcl-2、bcl-2 mRNA的阳性表达,抑制促凋亡因子bax、bax mRNA的阳性表达。

Objective It is to evaluate the effect of Gengnian decoction on ovarian tissue morphology,bax,bax mRNA,bcl-2,bcl- 2 mRNA which is one of apopcosis factor of Kidney-Yang deficiency type perimenopause rat＇ovarian granulosa cells.Methods To build natural aging perimenopause model rats by selecting the exfoliated cervicall cells of female SD rats on 11-14 months age and intramuscular injection altemately hydrocortisone to build the rat model with Kidney-Yang deficiency type perimenopause. Then the models were randomly divided into four groups： Gennian decoction group,textbook group,Livial group and normal saline group. At the same time,the study chose four-month-old female SD rats and eighteen-month-old female SD rats as compared group. The young group,elderly group and normal saline group were treated with normal saline by gavage,every treatment group were treated with respective drug by gavage. After treatment accordingly,by adopting pathological sections to understand morphological changes of rats＇ ovarian,adopting Immunohistochemistry and in Situ Hybridization technique to compare the positive expression of ovarian granulose cell bax,bax mRNA,bcl- 2,bcl- 2 mRNA in every group.Results Each dosage groups compared with normal saline group,the elderly group,follicular development situation were well（ P 〈0. 05）,follicular atresia,follicle expansion were reduced（ P 〈0. 05）; The Gengnian decoction and Livial group of follicular situation was superior to the textbooks party group（ P 〈0. 05）,and the difference between the two groups had no statistical significance（ P 〉0. 05）. The follicular atresia,follicle expansion in Gengnian decoction group were better than that in Livial group,textbooks group（ P 〈0. 05）,the textbook group compared with the Livial group had no statistical significance（ P 〉0. 05）. Each dosage groups compared with normal saline group,the elderly group,bax,bax mRNA positive expression were lower（ P 〈0. 05）,the bcl- 2 and bcl- 2 mRNA positive expression had raised（ P 〈0. 05）; The Gengnian Decoction and Livial group＇s bax,bax mRNA positive expression were lower than that in group teaching material side（ P 〈0. 05）,the bcl- 2 and bcl- 2 mRNA positive expression were significantly higher than textbooks party group（ P 〈0. 05）,there was no statistically significant difference between Gennian decoction and Livial group（ P 〈0. 05）. Conclusion Gengnian decoction can promote follicular development,inhibit follicular atresia and expansion,up-regulate the positive expression of bcl- 2 and bcl- 2mRNA,down-regulate the positive expression of bax and bax mRNA in ovarian granulosa cells.