SOCS3在脂多糖诱导体外氧糖剥夺大脑皮层细胞药理性预适应中的作用  

Anti-inflammatory Effect of SOCS3 in Preconditioning Induced by LPS in Cortical Cells Suffering from Oxygen-Glucose Deprivation Injury

作　　者：潘琳娜 樊萍 刘海云[2] 李建斌 宋志坚 马炜林 潘旭 吕青[3] Pan Linna;Fan Ping;Liu Haiyun(Medical Department of Neurology,Jiangxi Province Hospital of Integrated Chinese and Western Medicine,Jiangxi Traditional Chinese Medicine University Affiliated Chinese and Western Medicine Hospital,Nanchang 330003,China;Department of Physiology,Jiangxi Traditional Chinese Medicine University,Nanchang 330004,China)

机构地区：[1]江西省中西医结合医院(江西中医药大学附属中西医结合医院)神经内科,南昌330003 [2]江西中医药大学生理学系,南昌330004 [3]华中科技大学同济医学院基础医学院药理学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2020年第5期550-555,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81660600)。

摘　　要：目的观察脂多糖(lipopolysaccharides,LPS)药理性预适应对大脑皮层细胞缺糖缺氧损伤是否有保护作用,探讨在体外神经细胞中JAK2-STAT3-SOCS3信号通路与炎性通路之间是否相互调控,参与LPS药理性预适应过程,从而对大脑皮层细胞缺糖缺氧损伤起到保护作用。方法原代培养大鼠乳鼠大脑皮层神经细胞,氧糖剥夺(oxygen-glucose deprivation,OGD)损伤2 h再复氧糖24 h模拟缺血再灌注损伤过程,MTT检测细胞活力,乳酸脱氢酶(LDH)释放实验评价细胞损伤程度,观察LPS预处理48 h对体外大脑皮层神经细胞氧糖剥夺损伤的保护作用;ELISA法检测细胞培养上清中炎性因子IL-6、IL-1β、TNF-ɑ含量;Western blot检测大脑皮层细胞SOCS3、p-JAK2、JAK2、p-STAT3、STAT3表达情况。结果(1)与溶剂对照组(Sham组)比较,LPS+Sham组细胞活力及LDH释放无明显改变,提示0.02~0.05μg/mL LPS处理对神经细胞无明显损伤;LPS+Sham组细胞p-JAK2、p-STAT3、SOCS3蛋白表达有一定升高趋势,但差异无统计学意义(均P>0.05)。(2)与Sham组相比,OGD组细胞活力下降,LDH漏出量增多,培养上清中TNF-ɑ、IL-6、IL-1β含量增加,p-JAK2/JAK2比值、p-STAT3/STAT3比值、SOCS3表达量均明显升高(均P<0.05)。(3)与OGD组相比,低剂量LPS+OGD和高剂量LPS+OGD组细胞OGD损伤均减轻,细胞培养上清中炎性因子IL-6、IL-1β、TNF-ɑ分泌减少,细胞中SOCS3表达增加,而p-JAK2/JAK2比值、p-STAT3/STAT3比值明显降低(均P<0.05)。结论LPS药理性预适应能减轻大脑皮层神经细胞OGD损伤程度,这一神经保护作用可能通过JAK2-STAT3通路上调OGD神经细胞SOCS3蛋白表达,抑制炎性反应而实现。Objective To observe whether LPS-induced preconditioning has protective effects on cortical cells suffering from oxygen-glucose deprivation(OGD)injury and to explore whether the mechanism of the effect of LPS-induced preconditioning on cortical cells suffering from oxygen-glucose deprivation injury is related to JAK2-STAT3-SOCS3 and inflammatory pathway.Methods Lethal 2 h OGD followed by 24 h of resupplying oxygen sugar was designed as OGD group.Primary cortical cultures subjected to oxygen-glucose deprivation(OGD)were used as an in vitro simulated ischemic model.LPS was co-incubated with cortical cells 48 h before OGD.The toxicity of OGD was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium(MTT)assay and lactate dehydrogenase(LDH)leakage assay.The levels of inflammatory cytokines such as IL-6,IL-1βand TNF-αin cell culture medium were detected by ELISA kits.Western blot methods were used to detect the expression of proteins such as SOCS3,p-JAK2,JAK2,p-STAT3 and STAT3 in cortical cells.Results(1)Compared with the Sham group,the cells in"LPS+Sham"group with 0.02-0.05μg/mL LPS had no obvious damage,and the expression levels of proteins such as p-JAK2/JAK2,p-STAT3/STAT3 and SOCS3 were increased with no statistically significance(P>0.05).(2)Compared with the Sham group,the viability of the cortical cells in the OGD group decreased and the leakage of LDH increased.Meanwhile,the contents of IL-6,IL-1βand TNF-αin the culture medium were increased in the OGD group.Furthermore,The expression levels of p-JAK2/JAK2,p-STAT3/STAT3 and SOCS3 in the OGD group were increased significantly(P<0.05).(3)Compared with the OGD group,cell injury was alleviated in the low-dose and high-dose"LPS+OGD"group via inhibiting the secretion of inflammatory cytokines such as TNF-ɑ,IL-6 and IL-1βand promoting the expression of SOCS3 in cell culture medium.Meanwhile,the expression levels of p-JAK2/JAK2 and p-STAT3/STAT3 in the"LPS+OGD"group were significantly decreased(P<0.05).Conclusion(1)LPS-induced preconditioning is

关 键 词：脂多糖预适应 氧糖剥夺 炎性通路 SOCS3 

分 类 号：R743.3[医药卫生—神经病学与精神病学]