诃子鞣酸对LPS诱导的小鼠子宫内膜炎的保护作用  

Protective effect of chebulagic acid on LPS-induced endometritis in mice

作　　者：张兰欣 郭文瑞[1,2] 巩志国 李姣姣 王建宁 刘鑫煜 杨英 李政忆[1,2] 李娟 高瑞峰 ZHANG Lanxin;GUO Wenrui;GONG Zhiguo;LI Jiaojiao;WANG Jianning;LIU Xinyu;YANG Ying;LI Zhengyi;LI Juan;GAO Ruifeng(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot O10018,China;Key Laboratory of Clinical Diagnosis and Treatment Technology of Animal Diseases,Ministry of Agriculture and Rural Affairs,Hohhot 010018)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]农业农村部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2024年第5期1041-1048,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32160856);内蒙古自治区高等学校青年科技英才支持计划资助项目(NJYT24056);内蒙古自然科学基金资助项目(2023MS03035,2023LHMS03015)。

摘　　要：为探究诃子鞣酸(chebulagic acid, CA)对脂多糖(lipopolysaccharide, LPS)诱导的小鼠子宫内膜炎的保护作用。本研究体外试验部分,使用MTT法检测CA对小鼠腹腔巨噬细胞活性的影响,通过ELISA和Westen blot分别检测CA对LPS刺激的巨噬细胞促炎性细胞因子(TNF-α、IL-1β和IL-6)分泌水平和ERK、p38和p65磷酸化水平的影响;体内试验部分,通过ELISA分析CA对LPS刺激后小鼠子宫组织TNF-α、IL-1β和IL-6分泌水平的影响,使用组织免疫荧光法检测小鼠子宫损伤相关因子(HMGB1)的表达水平,通过HE染色法观察小鼠子宫组织的病理变化,使用血常规法分析小鼠全血中白细胞数、中性粒细胞数和单核细胞数的变化。结果显示,CA质量浓度为12.5、25.0、50.0 mg/L对巨噬细胞活性均无显著影响,因此后续试验结果与CA的细胞毒性无关。CA可显著下调受到LPS刺激的巨噬细胞中的TNF-α,IL-1β和IL-6分泌水平,且具有浓度依赖性。此外,CA可显著下调受LPS诱导的巨噬细胞中ERK、p38和p65的磷酸化水平。CA可显著下调LPS刺激小鼠后子宫组织中TNF-α、IL-1β和IL-6的分泌水平;LPS刺激可增加小鼠全血中的白细胞数、中性粒细胞数和单核细胞数,而使用CA治疗可使其显著降低;相比于LPS刺激组,CA治疗组子宫组织中HMGB1的表达水平显著降低;LPS刺激小鼠子宫后导致子宫角出血和水肿,使用CA治疗后子宫角出血程度和水肿程度均有所降低。同时,HE病理切片显示LPS刺激组小鼠子宫组织中出现大量的嗜中性粒细胞浸润,子宫内膜上皮发生增生、水肿、坏死及脱落,CA(100 mg/kg)治疗后,子宫组织基本恢复正常,具有浓度依赖性。以上结果表明,CA可通过抑制小鼠腹腔巨噬细胞中MAPK和NF-κB通路的激活,下调巨噬细胞和子宫组织中炎症介质的表达水平从而减轻LPS诱导的小鼠子宫内膜炎。This study aims to investigate the protective effect of chebulagic acid(CA)on lipopolysaccharide(LPS)-induced endometritis in mice.In the in vitro experiment part of this study,MTT assay was used to assess the effect of CA on the cell activity of mouse peritoneal macrophages.The effects of CA on the secretion of pro-inflammatory cytokines(TNF-α,IL-1βand IL-6)and the phosphorylation levels of ERK,p38 and p65 in LPS-stimulated macrophages were detected by ELISA and Western blot,respectively.In the in vivo test,the secretion levels of pro-inflammatory cytokines(TNF-α,IL-1βand IL-6)in the mouse uterine tissues after LPS stimulation and treatment by CA were analyzed by ELISA method,the expression level of mouse uterine injury-related factor(HMGB1)was detected by tissue immunofluorescence method,and the pathological changes of mouse uterine tissues were observed by HE staining.The changes of white blood cell count,neutrophil count and monocyte count in whole blood of mice were analyzed by routine blood test.The experimental results showed that the concentrations of CA at 12.5,25,50mg/L had no significant effect on macrophage activity,so the subsequent results were not related to the cytotoxicity of CA.CA significantly downregulated the levels of TNF-α,IL-1βand IL-6secretion in LPS-induced macrophages in a concentration-dependent manner.In addition,CA significantly downregulated the phosphorylation levels of ERK,p38,and p65in LPS-stimulated macrophages.CA significantly downregulated the expression levels of TNF-α,IL-1βand IL-6in uterine tissue after LPS stimulation in mice.LPS stimulation increased the number of leukocytes,neutrophils,and monocytes in whole blood of mice,while CA treatment significantly reduced the number of leukocytes.Compared with the LPS-stimulated group,the expression level of HMGB1in uterine tissue of the CA-treated group was significantly decreased.LPS stimulation of the mouse uterus resulted in hemorrhage and edema in the uterine horn,and the degree of hemorrhage and edema was reduced aft

关 键 词：子宫内膜炎 诃子鞣酸 脂多糖 炎症介质 炎症信号通路 高迁移率蛋白1 小鼠 

分 类 号：S859.7[农业科学—临床兽医学]