补肾健脾法对卵巢早衰小鼠抑制素B含量及基因表达的影响

Effect on Therapy of Tonifying Kidney and Strengthening Spleen on Inhibin B Content and Its Gene Expression in Premature Ovarian Failure Mice

【目的】观察补肾健脾法中药复方对卵巢早衰(POF)小鼠抑制素B(INHB)含量及基因表达的影响。【方法】选用雌性BALB/c小鼠随机分为正常对照组、模型组、中药预防组和中药治疗组。除正常对照组外,其他3组采用SD大鼠卵巢抗原与弗氏佐剂注射免疫法复制自身免疫性卵巢功能衰退小鼠模型。中药预防组于第1次免疫后开始灌服中药复方汤剂,连续17 d,中药治疗组于第3次免疫后开始灌服中药复方汤剂,连续5 d,剂量均为20 g.kg-1.d-1。采用酶联免疫吸附法检测各组血清INHB含量,逆转录—聚合酶链反应(RT-PCR)法检测各组卵巢组织INHB mRNA表达水平,并观察各组小鼠卵巢病理改变。【结果】模型组小鼠血清INHB含量、卵巢组织INHB mRNA相对表达量均显著降低(P<0.05或P<0.01),光镜下未见明显生长或成熟卵泡;中药预防组血清INHB含量、卵巢组织INHB mRNA相对表达量均显著升高(P<0.05或P<0.01),卵泡数目略减少,无明显病理改变;中药治疗组卵巢组织INHB mRNA相对表达量显著升高(P<0.05),而对血清INHB含量无影响(P>0.05),生长成熟卵泡减少,且卵巢组织出现病理改变。各组小鼠血清INHB含量与卵巢组织INHB mRNA表达变化呈相似趋势,且中药预防组作用优于中药治疗组。【结论】补肾健脾法中药复方治疗卵巢早衰的作用与其能增强卵巢组织INHB mRNA表达,升高INHB含量,改善卵巢的储备功能有关。

Objective To probe the mechanism of the Chinese compound recipe, which has the actions of tonifying the kidney and strengthening the spleen, on inhibin B （INHB） content and its gene expression level in mice with immune premature ovarian failure （POF）. Methods Female BALB/e mice were divided into 4 groups randomly： normal control group, model group, treatment group and prevention group. The autoimmune POF were induced in BALB/e mice except for the normal control group. Treatment group were treated with Chinese compound recipe （20 g·kg^-1·d^-1） for 5 continuous days after the third immunization, and prevention group were given Chinese compound recipe for 17 continuous days after the first immunization. The mice serum INHB content was detected by Sandwich enzyme linked immunosorbent assay （ELISA） method, and the ovarian INHB mRNA level was observed by reverse transcription polymerase chain reaction （RT-PCR） method. The ovarian pathological changes were also observed. Results The serum INHB content and ovarian INHB mRNA level were decreased obviously in the model group （P 〈 0. 05 or P 〈 0. 01 ）, and the microscopic examination results showed no grown or mature follicles. In the prevention group, serum INHB content and ovarian INHB mRNA level were increased obviously （P 〈 0. 05 or P 〈 0. 01 ）, the number of follicles was slightly reduced, but pathological changes were not shown in the ovarian tissue. In the treatment group, ovarian INHB mRNA level was increased （ P 〈 0.05 ）, but serum INHB content remained unchanged （P 〉 0. 05 compared with the model group）, the number of mature follicles was decreased and ovarian pathological changes were presented. The serum INHB content and ovarian INHB mRNA level showed the similar change after medication, and the effect in the prevention group was superior to that in the treatment group. Conclusion Chinese compound recipe with the actions of tonifying kidney and strengthening spleen can enhance the expression of ovarian INHB mRNA, increase the serum INHB content and improve the ovarian reserve function, which may be one of its mechanisms in treating POF.