摘要
目的探讨circ_0038467对缺氧缺糖(hypoxia-glucose deprivation,OGD)诱导的神经细胞损伤的影响及其可能作用机制。方法分离培养原代大鼠皮质神经细胞,采用OGD诱导大鼠皮质神经细胞建立细胞损伤模型;si-NC、si-circ_0038467、miR-NC、miR-940 mimics分别转染至大鼠皮质神经细胞后OGD处理6 h,si-circ_0038467和anti-miR-NC或anti-miR-940分别共转染至大鼠皮质神经细胞后OGD处理6 h;qRT-PCR法检测circ_0038467、miR-940的表达量;CCK-8法、流式细胞术分别检测细胞增殖及凋亡;LDH法检测细胞损伤情况;双荧光素酶报告实验用于靶向关系检测;Western blot检测cleaved caspase-3、cleaved caspase-9蛋白表达。结果OGD诱导的神经细胞中circ_0038467上调且miR-940下调(P<0.01);转染si-circ_0038467或miR-940 mimics后,细胞存活率升高(P<0.01),而LDH释放率、凋亡率和cleaved caspase-3、cleaved caspase-9水平下降(P<0.01);circ_0038467可靶向结合miR-940;相对于OGD+si-circ_0038467+anti-miR-NC组,细胞存活率在OGD+si-circ_0038467+anti-miR-940中降低(P<0.01),而LDH释放率、凋亡率和cleaved caspase-3、cleaved caspase-9水平增加(P<0.01)。结论干扰circ_0038467可上调miR-940表达保护神经细胞免受OGD诱导的氧化应激以及凋亡。
Aim To explore the effect of circ_0038467 on nerve cell damage induced by hypoxia-glucose deprivation(OGD)and its possible mechanism.Methods Rat cortical nerve cells were isolated and cultured,and then induced by OGD to establish a cell injury model.si-NC,si-circ_0038467,miR-NC,and miR-940 mimics were transfected into rat cortical nerve cells and treated with OGD for 6 h.si-circ_0038467 and anti-miR-NC or anti-miR-940 were co-transfected into rat cortical neurons,followed by OGD treatment for 6 h.qRT-PCR was used to detect the expression levels of circ_0038467 and miR-940.CCK-8 method and flow cytometry were used to examine cell proliferation and apoptosis.LDH method was used to detect cell damage.The dual luciferase reporter experiment was used to detect the targeting relationship between circ_0038467 and miR-940.Western blot was employed to detect cleaved caspase-3 and cleaved caspase-9 protein levels.Results Circ_0038467 expression increased and miR-940 expression decreased in OGD-induced nerve cells(P<0.01).After transfection with si-circ_0038467 or miR-940 mimics,cell survival rate increased(P<0.01),while LDH release rate,apopto-sis rate,and the protein levels of cleaved caspase-3 and cleaved caspase-9 decreased(P<0.01).Circ_0038467 could target miR-940.Compared with the OGD+si-circ_0038467+anti-miR-NC group,cell survival rate in OGD+si-circ_0038467+anti-miR-940 group was down-regulated(P<0.01),while LDH release rate,apoptosis rate and cleaved caspase-3,cleaved caspase-9 levels were up-regulated(P<0.01).Conclusion Interference of circ_0038467and could protect nerve cells from OGD-induced oxidative stress and apoptosis by up-regulating miR-940.
作者
孔炫栋
周黎琴
王宁
吴天涯
赵明
KONG Xuan-dong;ZHOU Li-qin;WANG Ning;WU Tian-ya;ZHAO Ming(Dept of Neurosurgery,Zhuji People’s Hospital of Zhejiang Province,Zhuji Zhejiang 311800,China;The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310051,China;Dept of Pharmacy,Zhuji People’s Hospital of Zhejiang Province,Zhuji Zhejiang 311800,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第5期887-893,共7页
Chinese Pharmacological Bulletin
基金
浙江省基础公益研究计划项目(No LGF22H090016)
浙江省医药卫生科技计划项目(No 2023RC108)
绍兴市卫生健康科技计划项目(No 2022KY100)。
