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目的:观察妇科养荣胶囊(Fu Ke Yang Rong capsule,FKYRC)对卵巢损伤小鼠卵巢储备功能和生育力的保护作用。方法:通过成年雌性小鼠一次性腹腔注射120 mg/kg环磷酰胺和10 mg/kg白消安建立卵巢损伤小鼠模型,于造模前7 d至造模后60 d每日对卵巢损伤小鼠用高、中、低剂量(6 g/kg、4 g/kg和2 g/kg)FKYRC(H组、M组、L组)连续灌胃给药;同时,于造模前7 d一次性皮下注射给予Gn RHa(38 mg/kg)作为阳性对照组(Gn RHa组),用生理盐水(0.2 m L/d)代替FKYRC连续灌胃作为卵巢损伤对照组(模型组),另设正常对照组:非卵巢损伤正常小鼠腹腔注射给予等体积DMSO(NC组)。分别于造模后30 d和60 d取材,计数卵巢组织中各级卵泡数量,检测血清E2、FSH、抑制素B(INHB)和抗苗勒氏管激素(AMH)水平,检测卵巢组织中翼状螺旋/叉头转录因子2(FOXL2)和AMH蛋白的表达水平。并观察造模60 d后各组妊娠率和窝仔数。结果:各给药组和模型组小鼠血清E2低于NC组(P<0.05),但FSH水平组间无统计学差异(P>0.05)。造模各组卵泡数明显低于NC组(P<0.05),且H组和Gn RHa组造模后60 d时卵巢组织中窦前卵泡与窦卵泡数较高,但各干预组与模型组比较无统计学差异(P>0.05)。模型组小鼠卵巢组织中FOXL2和AMH蛋白表达水平显著低于各FKYRC干预组(P<0.05),H组的妊娠率(80.00%)显著高于模型组(36.36%)(P<0.05)。结论:连续FKYRC(6 g/kg)给药60 d后可明显改善烷化剂所致卵巢损伤小鼠的卵巢储备功能和生育能力,其机制可能与上调颗粒细胞中FOXL2和AMH的表达水平相关。
Objective: To observe the protective effect of Fu Ke Yang Rong capsule (FKYRC) on ovarian reserve and fertility in ovariectomized mice. Methods: A mouse model of ovarian injury was established by intraperitoneal injection of 120 mg / kg cyclophosphamide and 10 mg / kg busulfan in adult female mice. Ovarian injury was induced daily from 7 days before model establishment to 60 days after model establishment Mice were administered intragastrically with high, medium and low doses of FKYRC (H group, M group and L group) at the doses of 6 g / kg, 4 g / kg and 2 g / kg respectively. Meanwhile, Gn RHa (38 mg / kg) was given as a positive control group (Gn RHa group) by subcutaneous injection once a day, and saline (0.2 m L / d) was used instead of FKYRC continuous gavage as control group (model group) Normal control group: normal ovariectomized mice were injected intraperitoneally with equal volume of DMSO (NC group). The number of follicles at each stage of ovarian tissue was counted at 30 d and 60 d after the model was established. The levels of E2, FSH, INHB and AMH in ovary tissue were measured. Spiral / Fork transcription factor 2 (FOXL2) and AMH protein expression levels. The pregnancy rate and litter size in each group were observed after 60 days. Results: Serum E2 was lower in NC group and model group than in NC group (P <0.05), but there was no significant difference between FSH group and control group (P> 0.05). The number of follicles in each model group was significantly lower than that in NC group (P <0.05), and the number of preantral follicles and antral follicles in ovarian tissue in H group and Gn RHa group at 60 d after modeling was higher than that in NC group No significant difference (P> 0.05). The expression of FOXL2 and AMH protein in ovary tissue of model group was significantly lower than that of FKYRC intervention group (P <0.05). The pregnancy rate (80.00%) in group H was significantly higher than that of model group (36.36%) (P <0.05). CONCLUSION: Continuous FKYRC (6 g / kg) administration for 60 d can significantly improve ovarian reserve and fertility in ovariectomized mice induced by alkylating agents. The mechanism may be related to the up-regulation of FOXL2 and AMH expression in granulosa cells .