摘要
目的:基于脂肪酸合酶(Fas)/脂肪酸合酶配体(FasL)信号通路探讨经皮穴位电刺激(TEAS)对早发性卵巢功能不全(POI)模型大鼠卵巢组织细胞凋亡的影响及可能机制。方法:54只雌性SD大鼠随机分为空白组15只及造模组39只。造模组以连续14 d雷公藤多苷灌胃方法构建POI大鼠模型,空白组以生理盐水灌胃作对照,造模结束随机处死3只大鼠取卵巢判断造模效果。造模成功后造模组大鼠被随机分为模型组、TEAS组及激素组,每组12只。TEAS组予每次20 min、连续4周电子针疗仪治疗,激素组连续0.09 mg/kg戊酸雌二醇灌胃治疗4周,同期空白组与模型组予生理盐水灌胃。治疗结束后,收集大鼠卵巢组织,酶联免疫吸附试验(ELISA)测定血清激素水平,脱氧核糖核苷酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法测定卵巢组织细胞凋亡率,蛋白质印迹法(Westernblotting)测定Fas、脂肪酸合酶相关死亡结构域蛋白(FADD)、胱天蛋白酶8(Caspase-8)蛋白表达。结果:与空白组相比,模型组血清雌二醇(E_2)水平显著降低(P<0.05),血清卵泡刺激素(FSH)、促黄体生成素(LH)水平、卵巢组织细胞凋亡水平和Fas、FADD、Caspase-8蛋白表达水平明显升高(P<0.05)。与模型组相比,TEAS组及激素组血清E_2水平显著升高(P<0.05),血清FSH、LH水平、卵巢组织细胞凋亡水平和卵巢组织Fas、FADD、Caspase-8蛋白表达水平明显降低(P<0.05)。结论:TEAS可以改善POI模型大鼠卵巢功能,其机制可能与调节血清性激素水平,下调Fas/FasL信号通路中关键效应分子Fas、FADD、Caspase-8蛋白表达,减少细胞凋亡有关。
Objective:Based on the fatty acid synthetase(Fas)/fatty acid synthetase ligand(FasL)signaling pathway,this study aims to investigate the effect and possible mechanism of transcutaneous acupoint electrical stimulation(TEAS)on apoptosis of ovarian tissue cells in rats with early-onset ovarian insufficiency(POI).Methods:A total of 54 female SD rats were randomly divided into a blank group(15 rats)and a modeling group(39 rats).The modeling group was used to construct a POI rat model by gavage of tripterygium wilfordii glycosides for 14 consecutive days.The blank group was used as a control group by gavage of normal saline.At the end of modeling,3 rats were randomly euthanized and their ovaries were taken to evaluate the modeling effect.After successful modeling,36 rats were randomly divided into model group,TEAS group and hormone group,12 rats each group.TEAS group was treated with an electronic acupuncture device for 20 minutes each time for 4 weeks.The hormone group were treated with 0.09 mg/kg estradiol valerate orally for 14 consecutive days.The blank group and the model group were treated with normal saline orally.After treatment,rat ovaries were collected.Serum hormone levels were measured using ELISA method,and the apoptosis rate of ovarian tissue cells was measured using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)method,and the expression of Fas,Fas-associated death protein with death domain(FADD),and Caspase-8 was measured using Western blotting.Results:Compared with the blank group,the serum level of estradiol(E2)in the model group was significantly reduced(P<0.05),while serum levels of follicle-stimulating hormone(FSH)and luteinizing hormone(LH),apoptosis level of ovarian tissue cell,and protein expression levels of Fas,FADD,and Caspase-8 were significantly increased(P<0.05).Compared with the model group,the serum level of E2 in each treatment group were significantly increased(P<0.05),while serum levels of FSH and LH,apoptosis levels of ovarian tissue cell,and protein expression levels of ovarian tissue Fas,FADD,and Caspase-8 were significantly reduced(P<0.05).Conclusions:TEAS can improve ovarian function in POI model rats,and its mechanism may be related to regulating serum sex hormone levels,downregulating the expression of key effector molecules Fas,FADD,and Caspase-8 proteins in the Fas/FasL signaling pathway,and reducing cell apoptosis.
作者
帅振虹
李天凤
李雪梅
SHUAI Zhenhong;LI Tianfeng;LI Xuemei(Center for Reproductive Health,Shenzhen Maternity and Child Healthcare Hospital of Southern Medical University,Shenzhen 518028,China)
出处
《山东中医药大学学报》
2025年第3期332-340,共9页
Journal of Shandong University of Traditional Chinese Medicine
基金
中国博士后科学基金项目(编号:2018M643206)
广东省基础与应用基础研究基金项目(编号:2020A1515110023)。
关键词
经皮穴位电刺激
早发性卵巢功能不全
脂肪酸合酶
脂肪酸合酶配体
脂肪酸合酶相关死亡结构域蛋白
胱天蛋白酶8
大鼠
transcutaneous acupoint electric stimulation
premature ovarian insufficiency
fatty acid synthetase(Fas)
fatty acid synthetase ligand(FasL)
Fas-associated protein with death domain(FADD)
Caspase-8
rat
作者简介
帅振虹,医学博士,副主任医师,主要从事生殖内分泌研究。邮箱:562599739@qq.com。
