制首乌对大鼠卵巢功能减退的影响

Effect of polygoni multiflori radix preparata on diminished ovarian reserve in rats

目的雌激素替代疗法治疗卵巢功能减退(DOR)的毒副作用较大。文中研究制首乌对大鼠卵巢功能减退的影响,为临床用药提供更优化的选择。方法将60只SD雌性大鼠随机数字表法分为正常组,模型组,制首乌高、中、低剂量组(4、2、1 g/kg)、阳性对照组(戊酸雌二醇0.18mg/kg)。每天上午10点各组均以雷公藤多甙(TG)75mg/kg灌胃造模(正常组灌服蒸馏水),下午5点制首乌各组和阳性对照组灌服相应药物,正常组和模型组灌服蒸馏水,连续30 d。观察大鼠动情周期,酶联免疫法检测大鼠血清雌二醇(E2)、卵泡刺激素(FSH)、黄体生成素(LH)、抗缪勒管激素(AMH)、抑制素B(INHB)的含量,计算大鼠子宫指数和卵巢指数,HE染色观察大鼠卵巢组织形态、计数各级卵泡。结果与正常组相比,模型组大鼠随造模时间的延长出现动情周期延长、不规则,在第11-30天阴道涂片显示为少量上皮细胞或少量角化细胞、持续白细胞;而制首乌各处理组与阳性对照组在第1-10天动情周期正常,在第11-30天出现动情周期相对延长,与模型组相比均有不同程度的恢复。与正常组E2含量[(302.6±42.9)pg/m L]、FSH含量[(7.2±0.5)m IU/m L]、LH含量[(17.4±1.2)m U/m L]比较,模型组大鼠血清E2含量[(155.7±46.8)pg/m L]明显降低(P<0.05),FSH、LH浓度[(21.7±1.2)m IU/m L、(25.0±1.0)m U/m L]显著升高(P<0.05);而血清FSH和LH浓度升高(P<0.05)。与正常组比较,模型组大鼠血清INHB含量明显降低[(494.5±84.1)pg/m L vs(299.2±106.8)pg/m L,P<0.05];制首乌高、中、低组和阳性对照组血清INHB水平[(595.2±66.3)、(741.8±94.1)、(644.3±92.0)、(659.1±84.7)pg/m L]升高(P<0.05)。与模型组相比,制首乌各组和阳性对照组INHB水平显著升高(P<0.05)。与正常组相比,模型组各级卵泡及黄体数量显著减少(P<0.05);制首乌各组和阳性对照组原始卵泡计数降低(P<0.05),生长卵泡计数增多(P<0.05),制首乌高剂量组闭锁卵泡+黄体计数升高(P<0.05),而制首乌低剂量组和阳性对照组闭锁卵泡+黄体计数降低(P<0.05)。与模型组相比,制首乌各组和阳性对照组原始卵泡计数和生长卵泡计数升高(P<0.01),制首乌高、中剂量组闭锁卵泡+黄体计数增加(P<0.05)。结论制首乌干预能抑制TG生殖毒性对大鼠卵巢的破坏,有效保护模型大鼠的生殖功能。

Objective Diminished ovarian reserve（ DOR） severely affects the life of women and the estrogen replacement therapy for it has obvious adverse effects. This article aimed to study the effect of polygoni multiflori radix preparata（ PMRP） on DOR in rats and provide a therapeutic option for clinical medication. Methods Sixty female SD rats were randomly divided into six groups of equal number,normal control,DOR model control,high-dose PMRP（ 4 g/kg）,medium-dose PMRP（ 2 g/kg）,low-dose PMRP（ 1 g/kg）,and positive control. The DOR model was established by gavage of tripterygium glycosides as 75 mg/kg every morning,followed by administration of PMRP in the PMRP groups,Estradiol valerate at 0.18 mg/kg in the positive control group and distilled water in the model control group in the afternoon,all for 30 consecutive days. The estrous cycle of the rats was observed,the levels of serum estradiol（ E2）,follicle-stimulating hormone（ FSH）,luteinizing hormone（ LH）,anti-Müllerian hormone（ AMH） and inhibin-B（ INHB） were determined by ELISA,the ovarian and uterine indexes were obtained,and the ovarian morphology was observed by HE staining,and the counts of follicles at different stages were recorded. Results Compared with the normal controls,the DOR model rats showed modeling time-related lengthening,irregularity and even disorder of the estrous cycle,with a few epithelial cells or keratinocytes and leucocytes on the vaginal smear at 11-30 days. The estrous cycle was normal in the PMRP and positive control groups at 1-10 days and relatively prolonged at 11-30 days. In comparison with the normal control group,the DOR model rats exhibited a significantly decreased levels of serum E2（ [302.6±42.9] vs [155. 7 ± 46. 8] pg/m L,P〈0. 05） and INH-B（ [494. 5 ± 84. 1] vs [299. 2 ±106.8] pg/m L,P〈0.05） but increased levels of FSH（ [7. 2 ± 0. 5] vs [21. 7 ± 1. 2] m IU/m L,P〈0. 05） and LH（ [17. 4 ± 1. 2] vs[25.0±1.0]m U/m L,P〈0.05）. The INH-B level was markedly elevated in the PMRP and positive control groups as compared with that in the DOR models（ P〈0.05）. The counts of follicles and corpora lutea were remarkably lower in the DOR model rats（ P〈0.05）while that of developing follicles markedly higher in the PMRP and positive control groups than in the normal control group（ P〈0.05）.The numbers of atretic follicles＋corpora lutea were significantly increased in the high-dose PMRP group but decreased in the low-dose PMRP group（ P〈0.05） and positive controls（ P〈0.05）. The counts of primordial and developing follicles were dramatically higher in the PMRP and positive control groups than in the DOR model controls（ P〈0.01） and so were the numbers of atretic follicles ＋ corpora lutea in the high-and medium-dose PMRP groups（ P〈0.05）. Conclusion Polygoni multiflori radix preparata can effectively protect the reproductive function of female rats by inhibiting tripterygium glycosides-induced toxicity to the ovary.