基于TLR2/MyD88/NF-κB信号通路探讨加味少腹逐瘀汤对寒湿瘀结证子宫内膜异位症痛经小鼠腹腔炎症微环境干预作用  

The intervention effect of Jiawei Shaofu Zhuyu decoction on peritoneal inflammatory microenvironment of dysmenorrhea mice with endometriosis of cold dampness and stasis syndrome based on TLR2/MyD88/NF-κB signaling pathway

作　　者：黄灿灿 毛海燕[2] 吉秀家 连小龙 张作良 陈元欢 岳斌 张小花 申剑 武权生 HUANG Can-can;MAO Hai-yan;JI Xiu-jia;LIAN Xiao-long;ZHANG Zuo-liang;CHEN Yuan-huan;YUE Bin;ZHANG Xiao-hua;SHEN Jian;WU Quan-sheng(Clinical School of Traditional Chinese Medicine,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Provincial People′s Hospital,Lanzhou 730030,China;the Department of Medical,Qinghai University,Xining 810016,China)

机构地区：[1]甘肃中医药大学中医临床学院,甘肃兰州730000 [2]甘肃省人民医院,甘肃兰州730000 [3]青海大学医学部,青海西宁810016

出　　处：《中国药理学通报》2024年第4期784-791,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82260949,81760879);甘肃省科技计划项目优秀博士生项目(No 23JRRA1220);甘肃中医药大学科学研究与创新基金项目(No 2021KCYB-2);甘肃中医药大学研究生创新创业基金(No 2022CX-01);西北环境与营养相关疾病中医药防控协同创新中心开放项目(No ZYXT-20-06)。

摘　　要：目的探讨加味少腹逐瘀汤对寒湿瘀结证子宫内膜异位症痛经小鼠腹腔炎症微环境的干预机制。方法构建动物模型后随机分组。以自主活动次数、粪便含水率、热痛潜伏期评价证候;以子宫及异位病灶大小评价疗效;HE染色比较病理变化;免疫组化检测异位组织中NF-κB的表达及其入核情况;qPCR法和Western blot测定病灶中TLR2、MyD88、NF-κB的mRNA和蛋白表达;ELISA检测TNF-α、IL-6、PGE2、E2、P。结果与假手术组对比,模型小鼠自主活动减少,热痛潜伏期缩短,粪便含水率增加;子宫体积增大,腹腔可见明显异位灶且HE染色显示炎症浸润;免疫组化显示NF-κB表达增加;异位组织TLR2、MyD88、NF-κB mRNA及蛋白表达增加,外周血TNF-α、IL-6、PGE2、E2增高,P下降。与模型组相比,各治疗组子宫体积减小、异位灶重量减轻、疼痛潜伏期延长;中药高剂量组小鼠自主活动次数增加、粪便含水率减少;药物干预后炎症反应改善、NF-κB表达下降;TLR2、MyD88、NF-κB的mRNA及蛋白表达降低;且外周血TNF-α、IL-6、PGE2、E2降低、P升高。结论加味少腹逐瘀汤可以降低寒湿瘀结证子宫内膜异位症小鼠血清炎症因子,这可能与抑制TLR2/MyD88/NF-κB信号通路改善腹腔炎症微环境有关。Aim To explore the intervention mechanism of Jiawei Shaofuzhuyu decoction on abdominal inflammatory microenvironment in mice with dysmenorrhea of cold dampness and blood stasis syndrome endometriosis.Methods The animal models were randomly grouped.Spontaneous activity times,fecal moisture content and heat pain latency were used to evaluate the syndrome.The efficacy was evaluated by the size of uterine and ectopic lesions.The pathological changes were compared using HE staining.The expression of NF-κB and its nucleation in ectopic tissues were detected by immunohistochemistry.The mRNA and protein expressions of TLR2,MyD88 and NF-κB were determined by qPCR and Western blot.TNF-α,IL-6,PGE2,E2 and P were detected by ELISA.Results Compared with sham operation group,the autonomic activity of model mice decreased,heat pain latency was shortened,and fecal moisture content increased.The uterine volume increased,the abdominal cavity showed obvious ectopic focus and HE staining showed inflammatory infiltration.Immunohistochemistry showed increased NF-κB expression.The mRNA and protein expressions of TLR2,MyD88 and NF-κB in ectopic tissues increased,TNF-α,IL-6,PGE2 and E2 in peripheral blood increased,and P decreased.Compared with the model group,the uterine volume,ectopic focus weight and pain latency were reduced in all treatment groups.The number of autonomous activities increased and fecal water content decreased in the high-dose group.After drug intervention,the inflammatory response was improved and the expression of NF-κB decreased.The mRNA and protein expressions of TLR2,MyD88 and NF-κB declined.In peripheral blood,TNF-α,IL-6,PGE2,E2 decreased and P increased.Conclusions Supplemented Shaofuzhuyu decoction can reduce serum inflammatory factors in mice with cold-dampness-stasis syndrome endometriosis,which may be related to inhibiting TLR2/MyD88/NF-κB signaling pathway to improve abdominal inflammatory microenvironment.

关 键 词：子宫内膜异位症 寒湿瘀结证 痛经 加味少腹逐瘀汤 TLR2/MyD88/NF-κB信号通路 炎症微环境 

分 类 号：R-332[医药卫生] R289.5R364.5R711.51R711.71