摘要
目的观察补肾健脾方对环磷酰胺诱导的卵巢储备功能减退大鼠模型卵巢功能的影响,并基于肿瘤坏死因子诱导相关细胞凋亡途径探讨补肾健脾方调控大鼠卵巢储备功能减退的机制研究。方法60只动情周期正常的健康雌性SD大鼠,随机分成正常组10只和造模组50只,造模组采用环磷酰胺75 mg/kg单次腹腔注射造模。造模成功的大鼠再随机分为模型组、西药组[戊酸雌二醇0.103 mg/(kg·d)+黄体酮胶囊2.575 mg/(kg·d)]、补肾健脾方低剂量组[3.21 g/(kg·d)]、补肾健脾方中剂量组[6.43 g/(kg·d)]、补肾健脾方高剂量组[12.85 g/(kg·d)],每组10只。正常组及模型组均予等体积蒸馏水灌胃,每组干预1次/d,干预14 d。检测各组大鼠动情周期、体质量、卵巢脏器指数、子宫脏器指数;ELISA法检测大鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、γ干扰素(interferon-γ,IFN-γ)含量;免疫荧光法检测大鼠卵巢颗粒细胞内B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)蛋白表达情况;HE染色检测卵巢组织病理情况;Western blot法检测卵巢磷酸化蛋白激酶B(posphorylated-proteinkinase B,p-AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mammalian target of rapamycin,p-mTOR)、自噬蛋白苄氯素-1(Beclin-1)表达情况。结果与正常组比较,模型组大鼠动情周期紊乱,可见卵巢结构紊乱、各级卵泡数量不同程度的减少、未见成熟卵泡,卵巢间质中见到明显炎细胞浸润;大鼠体质量、卵巢脏器指数和子宫脏器指数无明显变化;血清TNF-α、IFN-γ水平升高(P<0.05,P<0.01);卵巢颗粒细胞Bcl-2、Bax、Beclin-1蛋白表达水平升高(P<0.05,P<0.01),p-AKT、p-mTOR蛋白表达水平降低(P<0.01)。与模型组比较,西药组及补肾健脾方低、中剂量组血清TNF-α、IFN-γ水平降低(P<0.05,P<0.01);补肾健脾方中剂量组Bax蛋白表达水平降低(P<0.05);补肾健脾方高剂量组Bcl-2蛋白�
Objective To observe the effects of Bushen Jianpi Formula(BSJPF)on ovarian function in rats with cycloph-osphamide-induced decreased ovarian reserve function,and to investigate the mechanism of BSJPF regulating diminished ovarian reserve(DOR)in rats based on tumor necrosis factor-induced apoptosis pathway.Methods A total of 60 healthy female SD rats with normal estrous cycle were randomly divided into normal group(10 rats)and modeling group(50 rats).The modeling group was treated with a single intraperitoneal injection of cyclophosphamide at 75 mg/kg to establish models.The successful modeling rats were then randomly divided into model group,western medicine control group(estradiol valerate 0.103 mg/kg·d+progesterone capsule 2.575 mg/kg·d),and low-(3.21 g/kg·d),medium-(6.43 g/kg·d),and high-dose(12.85 g/kg·d)BSJPF groups,with 10 rats in each group.Normal group and model group were given equal volume of distilled water by gavage once a day,for 14 consecutive days.Estrous cycle,body mass,ovarian organ index,and uterine organ index were checked.The content of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)in serum of rats were determined by ELISA.The expressions of B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)in ovarian granulosa cells were measured by immunofluorescence assay.HE staining was used to examine the pathological status of ovarian tissues.Western blot analysis was used to check the ovarian posphorylated-proteinkinase B(p-AKT),p-mammalian target of rapamycin(p-mTOR),and Beclin-1 expressions.Results Compared with the normal group,the estrous cycle of the model group was disturbed,the ovarian structure was disturbed,the number of follicles at all levels was reduced to different degrees,no mature follicles were seen,and obvious inflammatory cell infiltration was seen in the ovarian stroma.There were no significant changes in body mass,ovarian organ index,or uterine organ index.Serum levels of TNF-αand IFN-γwere higher(P<0.05,P<0.01).The protein expressions of Bcl-2,Bax,and Becl
作者
周芳
曾晶
王永连
陈嘉盈
刘牧涯
ZHOU Fang;ZENG Jing;WANG Yonglian;CHEN Jiaying;LIU Muya(The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China)
出处
《湖南中医药大学学报》
CAS
2024年第7期1167-1174,共8页
Journal of Hunan University of Chinese Medicine
基金
长沙市自然科学基金项目(Kq2202452)
湖南省卫生健康委员会重点指导项目(C202305017704)
湖南省中医药管理局委托项目(D2023002)
湖南中医药大学校级重点项目(2019XJJJ036)
湖南中医药大学研究生创新课题校级项目(2023CX96)。
