PTPRO激活AKT促进脂多糖(LPS)诱导的小鼠急性肺损伤中肺泡上皮细胞的吞噬  

Protein tyrosine phosphatase receptor type O(PTPRO)promotes phagocytic activity of alveolar epithelial cells via activating AKT signal pathway in LPS induced acute lung injury

作　　者：董逸舒 韩雨红 陈晓芬 李雪蒙 熊倩倩 钱中清 李柏青 汪洪涛 吴凤娇 DONG Yishu;HAN Yuhong;CHEN Xiaofen;LI Xuemeng;XIONG Qianqian;QIAN Zhongqing;LI Baiqing;WANG Hongtao;WU Fengjiao(Anhui Province Key Laboratory of Immunology in Chronic Diseases,Bengbu Medical College,Bengbu 233030,China;Department of Immunology,School of Clinical Laboratory Medicine,Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]慢性疾病免疫学基础与临床安徽省重点实验室,安徽蚌埠233030 [2]蚌埠医学院检验医学院免疫学教研室,安徽蚌埠233030

出　　处：《细胞与分子免疫学杂志》2022年第12期1091-1096,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81801573,82071849);安徽省青年科学基金(1808085QH253);蚌埠医学院研究生科研创新计划(Byycx22017)。

摘　　要：目的探讨受体O型蛋白酪氨酸磷酸酶(PTPRO)在脂多糖(LPS)诱导的急性肺损伤中对肺泡上皮细胞吞噬功能的影响。方法体内实验将小鼠随机分成正常对照组和LPS刺激组,通过LPS腹腔注射建立急性肺损伤模型,HE染色检测小鼠肺组织中炎性细胞的浸润情况,ELISA检测小鼠肺组织中细胞因子肿瘤坏死因子α(TNF-α)的表达,Western blot法检测LPS对小鼠肺组织PTPRO蛋白表达的影响。体外实验利用小干涉RNA(siRNA)沉默MLE-12肺泡上皮细胞PTPRO的表达,流式细胞术检测LPS和PTPRO沉默对MLE-12细胞吞噬功能的影响;Western blot法检测LPS和PTPRO沉默对MLE-12细胞蛋白激酶B(AKT)和磷酸化AKT蛋白表达的影响。结果LPS腹腔注射建立小鼠急性肺损伤模型后,小鼠肺中浸润的多形核白细胞显著增多,肺组织中TNF-α的表达也升高。小鼠肺组织和MLE-12细胞的PTPRO蛋白表达量随LPS刺激在24 h内呈时间依赖性上调。PTPRO沉默后,MLE-12细胞的吞噬能力显著下降,且其AKT磷酸化水平的表达较对照组显著降低。结论PTPRO可能通过激活AKT信号通路促进MLE-12细胞的吞噬。Objective To investigate the effect of protein tyrosine phosphatase receptor type O(PTPRO)on the phagocytic activity of alveolar epithelial cells in LPS-induced acute lung injury.Methods Mice were randomly divided into the normal control group and LPS stimulation group.The infiltration of inflammatory cells was detected by HE staining.The cytokine TNF-αlevel in lung was analyzed by ELISA.Western blotting was performed to detect the effect of LPS on PTPRO protein expression in lung.After the expression of PTPRO in MLE-12 cells was silenced by siRNA in vitro,flow cytometry was used to detect the effects of LPS and PTPRO siRNA on the phagocytic activity of MLE-12 cells,and the effects of LPS and PTPRO siRNA on the expression of PTPRO,AKT and phosphorylated AKT protein were measured by Western blotting.ResultsAfter the establishment of murine acute lung injury model by LPS injection(1 mg/kg),the infiltrated polymorphonuclear leukocytes were markedly increased.The level of TNF-αin lung tissue and the expression of PTPRO in MLE-12 cells were both significantly increased after LPS stimulation.However,the activity of MLE-12 cells to phagocytose fluorescent microbeads was evidently decreased after silencing PTPRO.Furthermore,silencing PTPRO induced a remarkable decrease in the phosphorylation of AKT in MLE-12 cells.Conclusion PTPRO can promote phagocytic activity of MLE-12 cells via activating AKT signaling pathway.

关 键 词：PTPRO 急性肺损伤 肺泡上皮细胞 吞噬 

分 类 号：R563[医药卫生—呼吸系统] R563.1[医药卫生—内科学] R364.5[医药卫生—临床医学]