摘要
目的:探索毓麟珠调控叉头框蛋白O3a(FOXO3a)通路改善早发性卵巢功能不全(POI)小鼠卵巢功能,促进卵泡发育的作用机制。方法:选取BALB/c雌性小鼠皮下注射小鼠透明带3(Zp3)建立免疫POI模型,造模成功后随机分为模型对照组、阳性对照组(戊酸雌二醇0.009 mg/mL)、低剂量组(毓麟珠0.234 mg/mL)、高剂量组(毓麟珠0.936 mg/mL),每组10只,另设10只小鼠作为空白组。放射免疫法检测血清雌激素(E_(2))、促卵泡生长激素(FSH)、促黄体生成素(LH)和抗苗勒激素(AMH)水平,HE染色观察卵巢组织形态,Western Blot检测卵巢组织FOXO3a、过氧化物酶体增殖物激活受体-γ共激活因子-1α(PGC-1α)蛋白表达水平,实时荧光定量PCR检测卵巢组织FOXO3a、去乙酰化酶3(SIRT3)、去类泛素化特异性蛋白酶1(SENP1)mRNA水平。结果:阴道涂片观察显示,给药结束后,阳性对照药组、低剂量组、高剂量组动情周期基本恢复规律。POI小鼠卵巢分泌E_(2)、AMH显著减少(P<0.01),FSH、LH浓度显著升高(P<0.01)。与模型组比较,高剂量组E_(2)、AMH显著升高(P<0.01),FSH、LH浓度显著降低(P<0.01)。模型组小鼠FOXO3a表达呈上升趋势,PGC-1α蛋白表达呈下降趋势,给予中药后的POI小鼠FOXO3a蛋白表达呈下降趋势,PGC-1α蛋白表达呈升高趋势。POI模型小鼠FOXO3a、SIRT3、SENP1基因表达水平均显著升高(P<0.01),给予中药的低剂量组基因FOXO3a、SENP1表达有下降趋势,SIRT3显著下降(P<0.01)。结论:毓麟珠通过下调氧化诱导因子和线粒体乙酰化因子,上调抗氧化因子调节氧化还原反应,改善了卵母细胞线粒体功能,促进卵泡发育从而改善卵巢功能。
Objective:To explore the mechanism of Yu Linzhu regulating FOXO3a pathway to improve ovarian function and promote follicular development inpremature ovarian insufficiency(POI)mice.Methods:The immune POI model was established by subcutaneously in jecting mouse pellucida 3(Zp3)into female BALB/c mice.After successful modeling,the mice were randomly divided into model control group,positive control group(0.009 mg/mL estradiol valerate),low-dose group(0.234 mg/mL Yu Linzhu)and high-dose group(0.936 mg/mL Yu Linzhu),with 10 mice in each group.Another 10 mice were set as blank group.The levels of serum estrogen(E_(2)),follicle-stimulating growth hormone(FSH),luteinizing hormone(LH)and anti-mullerian hormone(AMH)were detected by radioimmunoassay.The morphological and pathological characteristics of ovarian follicles were observed by HE staining.The protein expression levels of FOXO3a and PGC-1αin ovarian tissues were detected by Western Blot.Real-time fluorescence quantitative PCR was used to detect FOXO3a,SIRT3 and SENP1 mRNA level in ovarian tissue.Results:Vaginal smear observation showed that after administration,estrus cycle of positive control group,low dose group and high dose group basically recovered.The ovaries of POI mice secreted E_(2) and AMH decreased(P<0.01),and the concentration of FSH and LH increased(P<0.01).Compared with the model group,E_(2),AMH increased(P<0.01),FSH,LH decreased in mice of high dose group of traditional Chinese medicine(P<0.01).FOXO3a expression showed increased trend and PGC-1αexpression showed decreased in model control mice,while FOXO3a expression showed decreased trend and PGC-1αexpression showed increased in POI mice after traditional Chinese medicine treatment.The expression levels of FOXO3a,SIRT3,SENP1 genes increased in POI model mice.FOXO3a,SENP1 and SIRT3 genes decreased in the low-dose group of traditional Chinese medicine(P<0.01).Conclusion:Yu Linzhu can regulate redox reaction by down-regulating oxidative inducible factor and mitochondrial acetylation factor,up-regulating antioxidant factor,improving mitochondrial function of oocytes,promoting follicular development and improving ovarian function.
作者
杨珍
魏茂林
董晓英
YANG Zhen;WEI Mao-lin;DONG Xiao-ying(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2022年第10期6026-6030,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81503607)
北京市自然科学基金项目(No.7212162)
北京中医药薪火传承“3+3”项目(No.2012-SZ-C41)。
