2,3,5,4'-Tetrahydroxystilbene-2-O-β-glucoside Alleviates PCOS-like Characteristics by Upregulating The Expression of CYP19A1 in Granular Cells of Sinus Follicles to Inhibit Inflammatory Response  

TSG通过上调窦状卵泡中颗粒细胞CYP19A1的表达抑制炎症反应缓解大鼠多囊卵巢综合征样特征

作　　者：YAO Jian-Feng XU Bo LIU Pin-Yue CHENG Di WEI Le PAN Xin-Yun MO Zhong-Cheng LI Ming 姚建凤;徐波;刘品月;成迪;魏乐;潘信运;莫中成;李明(泉州市妇幼保健院妇产科,泉州362000;湖南医药学院医学人文与信息管理学院,怀化418000;湖南医药学院基础医学院,怀化418000;桂林医学院基础医学院,桂林541199)

机构地区：[1]Department of Obstetrics and Gynecology,Quanzhou Maternity and Child Healthcare Hospital,Quanzhou 362000,China [2]Medical Humanity and Information Management College,Hunan University of Medicine,Huaihua 418000,China [3]School of Basic Medical Sciences,Hunan University of Medicine,Huaihua 418000,China [4]School of Basic Medical Sciences,Guilin Medical University,Guilin 541199,China

出　　处：《生物化学与生物物理进展》2025年第3期735-750,共16页Progress In Biochemistry and Biophysics

基　　金：湖南自然科学基金(2021JJ30482);怀化市科技计划(2021R3106);华侨大学联合创新项目(2021YX004);泉州市科技计划(2022C034R);广西研究生教育创新项目(YCSW2023406)资助。

摘　　要：Objective To investigate whether 2,3,5,4'-tetrahydroxystilbene-2-O-β-glucoside(TSG)ameliorated polycystic ovary syndrome(PCOS)-like characteristics by inhibiting inflammation.Methods PCOS models were established by injecting subcutaneously with dehydroepiandrosterone into female Sprague-Dawley rats,followed by receiving intraperitoneal injection of TSG.The granular cells(GCs)KGN were transfected with small interfering RNAs(si-NC and si-CYP19A1).The cells were preincubated with lipopolysaccharide(LPS)and then treated with or without TSG.The estrous cycle was monitored using vaginal exfoliated cells.The morphology of ovarian follicles was analyzed by H&E staining.ELISA was used to analyze estradiol(E2),testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),IL-6,TNF-α,AGEs,CRP and Omentin-1 levels in serum.Immunohistochemistry was performed to analyze PCNA and CYP19A1 expressions in the GCs of ovaries.Tunel staining was executed to detect the apoptosis of GCs.Quantitative polymerase chain reaction(qPCR)and Western blot were implemented to measure the expression of CYP19A1 in the ovaries and transfected cells.qPCR was used to analyze the expression of IL-6 and TNF-αin the transfected cells treated with LPS and TSG.Results The estrous cycles were restored in TSG group.Compared with model group,the sinus follicles were reduced and corpus luteums were increased in TSG group.TSG group showed increased E2,and decreased T and LH,compared with model group.Pro-inflammatory factors(IL-6,TNF-α,CRP and AGEs)were decreased,and anti-inflammatory factor(Omentin-1)was increased in TSG group compared with those in model group.TSG could partially inhibit decrease of PNCA-positive GCs and increase of Tunel-positive GCs caused by PCOS.The CYP19A1 expression of GCs in TSG group was upregulated compared with model group.The expressions of IL-6 and TNFαin si-CYP19A1 cells were increased compared with si-NC cells.Compared with cells(si-NC and si-CYP19A1)treated without LPS,the expressions of IL-6 and TNF-αcells wer目的为了探讨2,3,5,4'-四羟基二苯基-2-O-β-葡萄糖苷(TSG)是否通过抑制炎症改善多囊卵巢综合征(PCOS)样特征。方法皮下注射脱氢表雄酮建立雌性SD大鼠PCOS模型,然后腹腔注射TSG。颗粒细胞(GCs)KGN转染小分子干扰RNAs(si-NC和si-CYP19A1)后,先用脂多糖(LPS)预孵育,再用(/不用)TSG处理。阴道脱落细胞监测发情周期;苏木精-伊红(HE)染色分析卵巢卵泡形态;酶联免疫吸附分析(ELISA)法检测血清雌二醇(E2)、睾酮(T)、卵泡刺激素(FSH)、黄体生成素(LH)、IL-6、肿瘤坏死因子α(TNF-α)、晚期糖基化终末产物(AGEs)、C反应蛋白(CRP)、网膜素1(Omentin-1)水平;免疫组化法检测GCs中增殖细胞核抗原(PCNA)和芳香化酶(CYP19A1)的表达;原位末端标记(Tunel)染色检测GCs的凋亡情况;采用实时定量PCR(qPCR)和蛋白质免疫印迹(Western blot)检测卵巢和转染细胞的CYP19A1表达;qPCR分析LPS和TSG处理后转染细胞中IL-6和TNF-α的表达。结果TSG治疗的PCOS鼠恢复了动情周期。与PCOS相比,TSG治疗的PCOS鼠卵巢中窦状卵泡减少,黄体增多。与PCOS相比,TSG处理的PCOS鼠E2升高,T和LH降低;促炎因子(IL-6、TNF-α、CRP、AGEs)降低,抗炎因子(Omentin-1)升高。TSG能部分抑制PCOS引起的PCNA阳性GCs的减少和Tunel阳性GCs的增加。与PCOS相比,经TSG处理后GCs的CYP19A1表达上调。转染siCYP19A1的细胞中IL-6和TNF-α表达量与si-NC细胞比明显增加。转染细胞经LPS预孵育后,IL-6和TNF-α的表达明显增加,CYP19A1表达下调。与经LPS孵育的细胞相比,经LPS和TSG处理后,细胞IL-6和TNF-α表达量减少,而CYP19A1表达增加。与LPS和TSG处理转染si-NC细胞相比,转染si-CYP19A1的细胞经LPS和TSG处理后,IL-6和TNF-α表达量明显增加。结论TSG通过上调GCs中CYP19A1的表达抑制炎症反应来缓解PCOS样特征。

关 键 词：2 3 5 4'-tetrahydroxystilbene-2-O-β-glucoside granular cells CYP19A1 polycystic ovary syndrome inflammatory response 

分 类 号：R711.6[医药卫生—妇产科学] Q955[医药卫生—临床医学]