温阳化浊方改善高龄小鼠子宫内膜容受性的机制研究  

Mechanism of improving effect on the endometrial receptivity of aged mice by Wenyang Huazhuo Formula

作　　者：赵晓丽 戎蓓蕾 江楠 冯伟华 陈明丽 窦真 夏天[1] ZHAO Xiao-li;RONG Bei-lei;JIANG Nan;FENG Wei-hua;CHEN Ming-li;DOU Zhen;XIA Tian(First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300193,China)

机构地区：[1]天津中医药大学第一附属医院生殖中心,国家中医针灸临床医学研究中心,天津300193

出　　处：《中华中医药杂志》2022年第9期5338-5344,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81774351);天津市教委科研计划项目(No.2019KJ047)。

摘　　要：目的:观察温阳化浊方对高龄小鼠子宫内膜炎症和子宫内膜容受性的影响,并探索其改善高龄小鼠子宫内膜容受性的机制。方法:ICR雌性2月龄小鼠30只为对照组;ICR雌性8月龄小鼠60只,其中30只为炎性衰老组,另30只为衰老给药组,给予中药温阳化浊方灌胃4周。各组雌性小鼠与雄性小鼠合笼,于见栓4.5d(Dpc4.5)取子宫组织和外周血,ELISA、RT-PCR方法检测血清和子宫组织炎症因子、雌激素受体α(ERα)和孕酮受体(PR)的表达水平;免疫组化和荧光染色法检测子宫内膜核受体共激活因子(NCOAs)的表达量;Western Blot检测子宫内膜容受性标志蛋白LIF、HOXA10的表达量;台盼蓝染色观察胚胎植入数量。结果:与对照组比较,炎性衰老组小鼠外周血TNF-α、IL-6、IL-1α、NF-κB、IL-10、TGF-β含量均显著增加,子宫组织NF-κB、IL-1β、IL-6mRNA表达显著增加(P<0.01,P<0.05);PR、NCOA1和NCOA2表达水平显著降低(P<0.05,P<0.01);子宫内膜容受性标志蛋白HOXA10、LIF蛋白表达量,胚胎植入数量显著降低(P<0.01)。与炎性衰老组比较,衰老给药组小鼠外周血TNF-α、IL-6、IL-1α和NF-κB含量显著降低,抑炎因子IL-10、TGF-β含量显著升高,子宫NF-κB、IL-1β、IL-6 mRNA表达降低,IL-10 mRNA表达增加(P<0.01,P<0.05);子宫PR、NCOA1、NCOA2、HOXA10和LIF的蛋白表达量显著增加(P<0.05,P<0.01);胚胎着床数量显著增加(P<0.01)。结论:温阳化浊方明显降低衰老子宫的慢性炎症状态,提高衰老子宫的内膜容受性,可能与增加PR和NCOAs表达,增强孕酮对子宫内膜的调控有关。Objective: To analyze the effects of Wenyang Huazhuo Formula(WYHZ) on endometrial chronic inflammation, endometrial receptivity, and embryo implantation in aged mice, and to explore the mechanism of WYHZ on improving the endometrial receptivity of aged mice. Methods: ICR female mice were divided into three groups with 30 mice in each group: young group(M2) in which mice were 2-month old;older control group(M9) in which mice were 8-month old;older treatment group(M9+WYHZ) in which mice were 8-month old and given WYHZ by oral gavage for 4 weeks. All the female mice were mated with male ICR mice. Four and half days after presentation of vaginal plug, the female mice were euthanized by cervical dislocation, and uterine tissues and peripheral blood were taken for subsequent study. The serum levels of inflammatory cytokines were examined by ELISA. The local expression of inflammatory cytokines, estrogen receptor α(ERα) and progesterone receptor(PR) in uterus was examined by RT-PCR. Measured the expression of uterine nuclear receptor coactivators(NCOAs) by immunohistochemistry and immunofluorescence. The expression of the endometrial receptivity markers(LIF and HOXA10) was examined by Western Blot. Detected the embryo implantation sites through trypan blue staining. Results:Compared to the M2 group, the serum levels of inflammatory cytokines, including TNF-α, IL-6, IL-1α, NF-κB, IL-10, TGF-βwere significantly increased in M9 group, uterine mRNA expressions of NF-κB, IL-1β, and IL-6 in M9 group were significantly highered(P<0.01, P<0.05);uterine expression levels of PR, NCOA1 and NCOA2 were significantly reduced(P<0.05, P<0.01);the protein levels of endometrial receptive markers HOXA10, LIF and the number of implanted embryo sites were significantly reduced in M9 group(P<0.01). Compared to M9 group, the serum levels of TNF-α, IL-6, IL-1α and NF-κB in M9+WYHZ group were significantly reduced, and the levels of anti-inflammatory cytokines IL-10 and TGF-β were increased, the mRNA expression levels of NF-κB, IL-

关 键 词：生殖衰老 孕酮受体 核受体共激活因子 子宫内膜容受性 炎症因子 

分 类 号：R285.5[医药卫生—中药学]