脂多糖刺激人乳腺上皮细胞SYK及炎症因子的表达变化  

Study on the expression of spleen tyrosine kinase and inflammatory factors stimulated by lipopolysaccharide in human mammary epithelial cells

作　　者：张铭 仝洁 刘赛赛 卢兴浩 丁倩 陈云帆 张小楠 赵士弟 杨帆 陈传好 ZHANG Ming;TONG Jie;LIU Saisai;LU Xinghao;DING Qian;CHEN Yunfan;ZHANG Xiaonan;ZHAO Shidi;YANG Fan;CHEN Chuanhao(Department of Anatomy,Bengbu Medical University,Bengbu Anhui 233030,China;Key Laboratory of Basic and Clinical Cardiovascular and Cerebrovascular Diseases,Bengbu Medical University,Bengbu Anhui 233030,China)

机构地区：[1]蚌埠医科大学人体解剖学教研室,安徽蚌埠233030 [2]蚌埠医科大学心脑血管疾病基础与临床重点实验室,安徽蚌埠233030

出　　处：《蚌埠医学院学报》2024年第3期296-301,306,共7页Journal of Bengbu Medical College

基　　金：蚌埠医学院自然科学重点项目(2020byzd031);博士科研启动基金资助项目(bsdq202005)。

摘　　要：目的:探讨脂多糖(LPS)刺激人乳腺上皮细胞中脾酪氨酸激酶(spleen tyrosine kinase,SYK)表达变化及对炎症因子分泌的影响。方法:不同浓度LPS诱导人乳腺上皮细胞产生炎症反应,制备急性乳腺炎细胞模型;通过CCK-8法检测细胞增殖活性;采用实时荧光定量PCR技术确定LPS最适刺激时间、浓度及该条件下SYK、炎症因子mRNA表达水平;免疫荧光法检测LPS刺激人乳腺上皮细胞后SYK蛋白表达的变化。结果:CCK-8结果显示,25μg/mL LPS刺激1 h,人乳腺上皮细胞增殖活性无明显变化(P>0.05);实时荧光定量PCR结果显示,25μg/mL LPS刺激1 h后乳腺上皮细胞SYK基因表达最显著(P<0.05);LPS刺激后,白细胞介素(IL)-1β、IL-6、IL-8、肿瘤坏死因子α和NLRP3表均增高(P<0.05),siRNA敲低SYK基因后,IL-1β、IL-6、肿瘤坏死因子α和NLRP3表达均降低(P<0.05)。结论:抑制SYK基因可减少LPS诱导的乳腺炎细胞模型炎症因子释放,对于乳腺上皮细胞具有一定保护作用。Objective:To investigate the expression levels of spleen tyrosine kinase(SYK)induced by LPS in human mammary epithelial cells,and its effects on the gene expression of inflammatory cytokine.Methods:The inflammatory response in human mammary epithelial cells were induced by LPS to establish the acute lactation mastitis cell model.The cell proliferation activity was detected by CCK-8 assay.The real-time quantitative PCR was used to obtain the optimal stimulation time and concentration of SYK,and detected the expression levels of related inflammatory cytokine genes.The expression levels of SYK genes in human mammary epithelial cells before and after LPS stimulation were detected by immunofluorescence.Results:The results of CCK-8 showed that there was no significant change in the proliferative activity of human mammary epithelial cells stimulated by 25μg/mL LPS for 1 h(P>0.05).The results of real-time fluorescence quantitative PCR showed that the expression of SYK gene in mammary epithelial cells was the most significant after 25μg/mL LPS stimulation for 1 h(P<0.05).After LPS stimulation,the expression levels of interleukin(IL)-1β,IL-6,IL-8,tumor necrosis factorαand NLRP3 increased(P<0.05),and the expression levels of IL-1β,IL-6,tumor necrosis factorαand NLRP3 decreased after the SYK gene knockdown by siRNA(P<0.05).Conclusions:Inhibition of SYK gene can reduce the release of inflammatory factors in LPS-induced mastitis cell models,and has a certain protective effect on mammary epithelial cells.

关 键 词：乳腺炎 脾酪氨酸激酶 脂多糖 炎症因子 

分 类 号：R655.8[医药卫生—外科学]