TLR7基因沉默通过NF-κB炎症通路对缺氧缺糖再给氧诱导的大鼠海马神经细胞凋亡的影响  

Effect of TLR7 gene silencing on the apoptosis of hippocampal neuronal induced by hypoxia, hypoglycemia and reoxygenation via NF-κB inflammatory pathway in rats

作　　者：程日霞 单伟 CHENG Ri-xia;SHAN Wei(Department of Anatomy,Basic Medical College,Jinzhou Medical University,Jinzhou 121000,China;Department of Neurology,Fukuang general hospital,Huarun liaojian group,Fushun 113008,China)

机构地区：[1]锦州医科大学基础医学院解剖学教研室,辽宁锦州121001 [2]华润辽健集团抚矿总医院神经内科,辽宁抚顺113008

出　　处：《解剖科学进展》2021年第4期486-488,492,共4页Progress of Anatomical Sciences

基　　金：辽宁省教育厅科学基金(L2013332);辽宁省自然科学基金指导计划(201602274)。

摘　　要：目的探讨Toll样受体7(Toll-like receptor 7, TLR7)基因对缺氧缺糖再给氧诱导的大鼠海马神经细胞凋亡的影响及机制。方法根据TLR7 mRNA编码序列设计并合成干扰siRNA序列,转染入大鼠海马神经细胞中,实验分为4组:对照组(Control)、缺氧缺糖再给氧模型组(Model)、TLR7基因沉默组(Model+TLR7-siRNA)、转染阴性对照组(Model+NC)。qRT-PCR检测沉默TLR7基因情况,Hoechst33258染色及流式细胞仪检测细胞凋亡率,Western blot检测TLR7、NF-κB、Caspase-3蛋白相对表达量。结果与Control组相比,Model组和NC-TLR7组TLR7基因及蛋白表达水平明显增加,Caspase-3、NF-κB蛋白表达水平及细胞凋亡率明显增加;与Model组相比,Model+TLR7-siRNA组TLR7基因及蛋白表达水平明显降低,Caspase-3、NF-κB蛋白表达及细胞凋亡率明显降低(P <0.05)。结论沉默缺氧缺糖再给氧诱导的大鼠海马神经细胞TLR7基因抑制细胞的凋亡,与下调Caspase-3、NF-κB表达有关。Objective To investigate the effect and mechanism of TLR7 gene on the apoptosis of hippocampal neuronal induced by hypoxia, hypoglycemia and reoxygenation in rats. Methods The interference siRNA sequence was designed and synthesized, and then transfected into rat hippocampal neurons. The experiment was divided into: Control, Model, Model+TLR7-siRNA and Model+NC groups. TLR7 gene silencing was detected by qRT-PCR, the apoptotic rate was detected by Hoechst33258 staining and flow cytometry, and the relative expression of TLR7, NF-κB and Caspase-3 was detected by Western blot. Results Compared with the control group, the expression level of TLR7 gene and protein increased significantly, the expression levels of Caspase-3 and NF-κB and apoptotic rate were increased significantly in Model and Model+NC groups. Compared with Model group, the expression level of TLR7 gene and protein decreased significantly, while the expression levels of Caspase-3 and NF-κB and apoptotic rate decreased significantly in Model+TLR7-siRNA group. Conclusion Silencing TLR7 gene inhibits the apoptosis of hippocampal neurons induced by hypoxia, hypoglycemia and reoxygenation, which is related to down-regulation of Caspase-3 and NF-κB expression.

关 键 词：缺血再灌注性脑损伤 基因沉默 TLR7基因 免疫炎症 凋亡 大鼠 

分 类 号：R743.31[医药卫生—神经病学与精神病学]