摘要
该文探讨了CircOGDH通过mi R-24-3p靶向调控HOXA1对缺氧复氧(H/R)诱导的神经元损伤的作用机制。以Ht22细胞为研究对象,利用缺氧复氧诱导神经元细胞损伤,将细胞分为阴性对照组(si-NC组)、CircOGDH沉默组(si-CircOGDH组)、过表达阴性对照组(miR-NC组)、miR-24-3p过表达组(miR-24-3p minic组)、CircOGDH沉默+抑制剂阴性对照组(si-CircOGDH+anti-miRNC组)、CircOGDH沉默+miR-24-3p抑制剂组(si-CircOGDH+anti-miR-24-3p组),另设置未转染的对照组(control组)、H/R组。qRT-PCR法检测CircOGDH、miR-24-3p、HOXA1 mRNA表达水平;CCK-8法检测细胞活性;氧化应激水平检测采用微量法;流式细胞仪检测细胞凋亡;Western blot检测HOXA1、Bax、Bcl-2蛋白表达。结果显示,H/R组Ht22细胞CircOGDH、HOXA1表达上调,miR-24-3p表达下调,细胞凋亡率以及Bax、LDH、MDA含量升高,细胞存活率以及Bcl-2水平、SOD活性、GSH-Px活性下降(P<0.05);沉默CircOGDH可以上调H/R诱导的Ht22细胞中miR-24-3p表达,下调HOXA1表达,提高细胞存活率和Bcl-2蛋白、SOD、GSH-Px水平,降低细胞凋亡率以及Bax、LDH、MDA含量(P<0.05);过表达miR-24-3p能够下调HOXA1表达,提高细胞存活率及以Bcl-2、SOD、GSH-Px水平,降低细胞凋亡率及以Bax、LDH、MDA含量(P<0.05);抑制miR-24-3p表达能够一定程度上逆转沉默CircOGDH对HT22细胞损伤的保护作用。由此提示,沉默CircOGDH可能通过上调miR-24-3p表达,下调HOXA1表达,改善H/R诱导的神经元氧化损伤,抑制其凋亡。
This study aimed to investigate the mechanism of CircOGDH on hypoxic reoxygenation(H/R)induced neuronal damage by targeting and regulating HOXA1 through miR-24-3p.Ht22 cells were used as the research object,hypoxia reoxygenation was used to induce neuronal damage,and the cells were divided into negative control group(si-NC group),CircOGDH silencing group(si-CircOGDH group),overexpression negative control group(miR-NC group),miR-24-3p overexpression group(miR-24-3p minic group),CircOGDH silencing+inhibitor negative control group(si-CircOGDH+anti-miR-NC group),and CircOGDH silencing+miR24-3p inhibitor group(si-CircOGDH+anti-miR-24-3p group).Control group and H/R group without transfection were also set up.The expression of CircOGDH,miR-24-3p and HOXA1 mRNA was detected by qRT-PCR;the cell activity was detected by CCK-8 method;the oxidative stress level was detected by micro method;apoptosis was detected by flow cytometry;Western blot was used to detect the expression of HOXA1,Bax and Bcl-2 proteins.The results showed that the expression of CircOGDH and HOXA1 was up-regulated and the expression of miR-24-3p was down-regulated in Ht22 cells in H/R group,the apoptosis rate,the expression of Bax protein,and the contents of LDH and MDA increased,the cell survival rate,the expression of Bcl-2 protein,the activities of SOD and GSH-Px decreased(P<0.05);silencing CircOGDH could up-regulate the expression of miR-24-3p and down-regulate the expression of HOXA1 in H/R-induced Ht22 cells,improve cell survival rate and the levels of Bcl-2 protein,SOD,and GSH-Px,reduce the apoptosis rate,Bax protein,the contents of LDH,MDA(P<0.05);overexpression of miR-24-3p could reduce the expression of HOXA1,increase the cell survival rate,the levels of Bcl-2 protein,SOD,GSH-Px,and reduce the apoptosis rate,Bax protein,contents of LDH and MDA(P<0.05);inhibiting the expression of miR-24-3p could partially reverse the protective effect of silencing CircOGDH on HT22 cells damage.In conclusion,silencing CircOGDH may up-regulate the expression of miR-24-3p,downregulate the expression of HOXA1,improve the oxidative damage of neurons induced by H/R,and inhibit their apoptosis.
作者
陈玉兰
陈萌
朱正萍
CHEN Yulan;CHEN Meng;ZHU Zhengping(Fever Clinic,Nanjing Pukou District TCM Hospital,Nanjing 211800,China;Department of Neurology,Nanjing Pukou District TCM Hospital,Nanjing 211800,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2023年第6期854-864,共11页
Chinese Journal of Cell Biology
基金
南京市浦口区社会事业科技发展计划(批准号:S2021-6)资助的课题。
