沉默HMGB1通过抑制p38 MAPK信号通路减少LPS或IL-17A诱导的中耳腔上皮细胞的炎症与凋亡

扫码查看

原文链接

万方数据
维普

中文摘要：目的:探讨高迁移率族蛋白B1(High Mobility Group Protein 1，HMGB1)对脂多糖(lipopo-lysaccharide，LPS)或白细胞介素-17A(interleukin-17A，IL-17A)诱导人中耳腔上皮细胞(human middle ear epithelial cells，HMEEC)的炎症反应与凋亡的调控作用与机制。方法:培养 HMEEC 细胞系。将HMEEC分为对照组、LPS组、LPS联合短发夹RNA(shRNA)沉默质粒阴性对照处理组(LPS+shNC 组)、LPS联合shRNA沉默HMGB1 处理组(LPS+shHMGB1 组)、LPS 联合 p38-丝裂原激活的蛋白激酶(p38 MAPK)抑制剂SB202190 处理组(LPS+SB202190 组)、IL-17A 组、IL-17A+shNC 组、IL-17A+shHMGB1组、IL-17A+SB202190 组。酶联免疫吸附测定(enzyme-linked immunosorbent assay，ELISA)法检测TNF-α、IL-1β和IL-6 的水平变化。用Western blot检测粘蛋白 5AC(mucoprotein 5AC，MUC5AC)、粘蛋白 8(mucoprotein 8，MUC8)、p38 MAPK、磷酸化的 p38 MAPK(p-p38 MAPK)、E26 样蛋白 1(E-twenty-six like 1 protein，ELK1)、B细胞淋巴瘤2 蛋白(B-cell lymphoma 2 protein，Bcl-2)，以及Bcl-2 相关X蛋白(Bcl-2 associated X protein，Bax)的表达。流式细胞术检测细胞的凋亡。结果:与对照组比，LPS组或IL-17A组的 HMEEC 的凋亡率都增加，且 HMGB1、MUC5AC、MUC8、TNF-α、IL-1β、IL-6、p38、p-p38、ELK1、Bax的表达水平升高，而 Bcl-2 的表达水平降低(均 P<0。05)。与 LPS 组或 IL-17A 组比，LPS+shHMGB1 组或IL-17A+shHMGB1 组的凋亡率都降低，且 HMGB1、MUC5AC、MUC8、TNF-α、IL-1β、IL-6、p38、p-p38、ELK1、Bax的表达水平都减少，而Bcl-2 的表达水平升高(均P<0。05)。与LPS 组或IL-17A组比，LPS+SB202190 组或 IL-17A+SB202190 组的的凋亡率都降低，MUC5AC、MUC8、TNF-α、IL-1β、IL-6、p38、p-p38、ELK1、Bax的表达水平都降低，而 Bcl-2 的表达水平升高(均 P<0。05)(均 P<0。05)。结论:沉默HMGB1 通过抑制p38 MAPK信号通路减少 LPS 或 IL-17A 诱导的 HMEEC 的炎症与凋亡。

外文标题：Silent HMGB1 Alleviates LPS or IL-17A Induced Inflammation and Apoptosis of Human Middle Ear Epithelial Cells via Inhibiting p38 MAPK Signaling Pathway

外文摘要：Objective:To investigate the regulatory effects and mechanisms of high-mobility group pro-tein B1(HMGB1)on the inflammatory response and apoptosis of human middle ear epithelial cells(HMEECs)induced by lipopolysaccharide(LPS)or interleukin-17A(IL-17A).Methods:HMEEC cell line was cultured.HMEECs were divided into the control group,LPS group,LPS+shRNA negative control group(LPS+shNC group),LPS+shRNA-mediated HMGB1 knockdown group(LPS+shHMGB1 group),LPS+p38 mitogen-activated protein kinase(p38 MAPK)inhibitor SB202190 group(LPS+SB202190 group),IL-17A group,IL-17A+shNC group,IL-17A+shHMGB1 group,and IL-17A+SB202190 group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the changes in the levels of TNF-α,IL-1β and IL-6.Western blot was used to detect the expression of mucin 5AC(MUC5AC),mucin 8(MUC8),p38 MAPK,phosphorylated p38 MAPK(p-p38 MAPK),E-twenty-six like 1 protein(ELK1),B-cell lympho-ma 2 protein(Bcl-2),and Bcl-2 associated X protein(Bax).Flow cytometry was used to detect cell apop-tosis.Results:Compared with the control group,the apoptosis rate of HMEECs in the LPS group or IL-17A group was increased,and the expression levels of HMGB1,MUC5AC,MUC8,TNF-α,IL-1β,IL-6,p38,p-p38,ELK1,and Bax were increased,while the expression level of Bcl-2 was decreased(all P<0.05).Compared with the LPS group or IL-17A group,the apoptosis rate was decreased in the LPS+shHMGB1 group or IL-17A+shHMGB1 group,and the expression levels of HMGB1,MUC5AC,MUC8,TNF-α,IL-1β,IL-6,p38,p-p38,ELK1,and Bax were all decreased,while the expression level of Bcl-2 was increased(all P<0.05).Compared with the LPS group or IL-17A group,the apoptosis rate was decreased in the LPS+SB202190 group or IL-17A+SB202190 group,and the expression levels of MUC5AC,MUC8,TNF-α,IL-1β,IL-6,p38,p-p38,ELK1,and Bax were all decreased,while the expression level of Bcl-2 was in-creased(all P<0.05).Conclusion:Silent HMGB1 alleviates LPS or IL-17A-induced inflammation and ap-optosis of HMEECs via inhibiting p38 MAPK signaling pathway.

外文关键词：

High mobility group protein 1p38 mitogen-activated protein kinaseHuman middle ear epithelial cellsCell apoptosis

作者：

阿不拉江·托合提、阿布利克木·依、吾买尔·亚生、韩志国

展开 >

作者单位：

新疆维吾尔自治区人民医院耳鼻咽喉诊疗中心,新疆乌鲁木齐 830001

关键词：

高迁移率族蛋白B1 p38-丝裂原激活的蛋白激酶人中耳腔上皮细胞细胞凋

基金：

新疆维吾尔自治区自然科学基金新疆维吾尔自治区人民医院院内项目

项目编号：

2022D01C13520200116

出版年：

2024

DOI：

10.3969/j.issn.1006-6233.2024.05.05

河北医学

河北省医学会

河北医学

CSTPCD

影响因子：1.915

ISSN：1006-6233

年,卷(期)：2024.30(5)

参考文献量15