TRPM7在七氟烷预处理抑制大鼠氧糖剥夺海马神经元凋亡和炎症反应中的作用

Role of TRPM7 in sevoflurane preconditioning for inhibiting hippocampal neurons apoptosis and inflammatory responses induced by oxygen-glucose deprivation in rat

目的探讨瞬时受体电位通道M7(TRPM7)在七氟烷(Sev)预处理抑制大鼠氧糖剥夺(OGD)海马神经元凋亡和炎症反应中的作用。方法选择出生1d的SD大鼠50只,提取海马神经元,将其分为5组,包括对照组、Sev预处理组(Sev组)、OGD组、Sev预处理+OGD组(Sev+OGD组)和Sev预处理+缓激肽+OGD组(联合组)。缺糖缺氧1.5h后复糖复氧,再正常培养24h后制备OGD模型。对照组海马神经元仅做正常培养;Sev组海马神经元行2%Sev预处理1h;OGD组海马神经元仅制备OGD模型;Sev+OGD组神经元行2%Sev预处理1h,24h后制备OGD模型;联合组神经元Sev预处理同时在培养基中加入缓激肽(终浓度200μmol/L),其余处理同Sev+OGD组。正常培养24h时后,检测海马神经元TRPM7 mRNA和蛋白表达水平、存活率和凋亡率、IL-1β、TNF-α的mRNA及上清蛋白表达水平。结果与对照组比较,OGD组海马神经元TRPM7 mRNA及蛋白表达水平、凋亡率、IL-1β、TNF-α mRNA及上清蛋白表达水平明显升高(P<0.05),而存活率明显降低(P<0.05)。与OGD组比较,Sev组海马神经元TRPM7 mRNA及蛋白表达水平、凋亡率、IL-1β、TNF-α mRNA及上清蛋白表达水平明显降低(P<0.05),而存活率明显升高(P<0.05)。与Sev组比较,联合组海马神经元TRPM7 mRNA及蛋白表达水平、凋亡率、IL-1β、TNF-α mRNA及上清蛋白表达水平明显升高(P<0.05),而存活率显著降低(P<0.05)。结论 Sev预处理可通过缓解神经元TRPM7过度表达,减轻OGD后海马神经元凋亡和炎症反应。

Objective To investigate the role of transient receptor potential melastatin 7（TRPM7）in sevoflurane preconditioning for inhibiting hippocampal neurons apoptosis and inflammation response induced by oxygen-glucose deprivation（OGD）.Methods Fifty SD rats of postnatal 1 dwere selected for extracting hippocampal neurons and randomly divided into 5 groups,including the control group（C）,sevoflurane preconditioning group（Sev）,OGD group,Sev preconditioning＋OGD group（Sev＋OGD）and Sev preconditioning＋bradykinin＋OGD group（combined group）.After 1.5 hoxygen-glucose deprivation,reintroduction was performed,and then the normal culture was performed again for preparing the OGD model.Hippocampal neurons in the control group were normally cultured only;which in the Sev group conducted 2% Sev preconditioning for 1 h;which in the OGD group only prepared the OGD model;which in the SEv＋OGD conducted 2% Sev preconditioning for 1 h,and prepared the OGD model after 24 h;which in the combined group was simultaneously added with bradykinin（final concentration 200μmol/L）in Sev preconditioning,other treatment was same to that in the Sev＋OGD group.After 24 hnormal culture,the mRNA and protein levels of TRPM7,apoptosis rate,survival rate,mRNA and supernatant protein levels of IL-1β and TNF-αof the hippocampal neurons were detected.Results Compared with the control group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,mRNA and supernatant protein levels of IL-1β and TNF-αin the OGD group were significantly increased（P〈0.05）,whereas the survival rate was significantly decreased（P〈0.05）.Compared with the OGD group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,mRNA and supernatant protein levels of IL-1β and TNF-αin the Sev group were significantly decreased（P〈0.05）,whereas the survival rate was significantly increased（P〈0.05）.Compared with the Sev group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,the mRNA and supernatant protein levels of IL-1β and TNF-αin the combined group were significantly increased（P〈0.05）,whereas the survival rate was significantly decreased（P〈0.05）.Conclusion Sev preconditioning can attenuate hippocampal neurons apoptosis and inflammatory response after OGD via alleviating the overexpression of TRPM7.