基于pgrmc1调控的黄体酮抑制氧糖剥夺/复氧新生小鼠神经元损伤机制分析OACSTPCD
Mechanism of progesterone in inhibiting neuron damage of neonatal mice subjected to OGD/R based on the regulation of pgrmc1
目的 基于黄体酮膜受体组件1(pgrmc1)调控,探讨黄体酮抑制氧糖剥夺/复氧(OGD/R)新生小鼠神经元损伤的作用机制.方法 选用出生12 h内的新生小鼠分离原代皮层神经元细胞,体外培养7 d,利用微管相关蛋白2检测进行鉴定后,随机分为对照组、二甲基亚砜(DMSO)组、AG205组、黄体酮组、AG205+黄体酮组.DMSO组、AG205组、黄体酮组、AG205+黄体酮组加入制备好的缺糖D-Hanks液、置于缺氧培养箱中培养2 h,换为神经元生长培养基;DMSO组在造模前1 h给予DMSO预处理,AG205组和AG205+黄体酮组在造模前1 h给予AG205(pgrmc1拮抗剂)10 μmol/L预处理,黄体酮组和AG205+黄体酮组于造模后2 h给予黄体酮20 μmol/L.复氧24 h后,采用CCK-8法检测细胞活力,流式细胞术检测凋亡细胞.结果 DMSO组、AG205组细胞存活率低于对照组,AG205组低于DMSO组;黄体酮组、AG205+黄体酮组细胞存活率高于AG205组,黄体酮组高于AG205+黄体酮组(P均<0.05).DMSO组、AG205组细胞凋亡率高于对照组,AG205组高于DMSO组,黄体酮组、AG205+黄体酮组细胞凋亡率低于AG205组(P均<0.05).结论 黄体酮可抑制OGD/R新生小鼠神经元损伤,抑制pgrmc1可降低OGD/R神经元活力、增加细胞凋亡,黄体酮抑制OGD/R神经元损伤的作用可能与调控pgrmc1有关.
Objective Based on the regulation of progesterone membrane receptor component 1(pgrmc1),to ex-plore the inhibition mechanism of progesterone(PROG)on neuronal damage of newborn mice subjected to oxygen glucose deprivation/reoxygenation(OGD/R).Methods Primary cortical neuronal cells were isolated from newborn mice within 12 hours of birth,cultured in vitro for 7 days,and were identified using microtubule-associated protein 2(MAP2)detec-tion.The neuronal cells were randomly divided into the control group,dimethyl sulfoxide(DMSO)group,AG205 group,PROG group,and AG205+PROG group,respectively.Neuronal cells in the DMSO group,AG205 group,PROG group,and AG205+PROG group were added with prepared glucose deficient D-Hanks solution and were cultured in an hypoxic incubator for 2 h,then were replaced with neuronal growth medium.At 1 h before OGD/R models were established,cells in the DMSO group were treated with DMSO,and cells in the AG205 and AG205+PROG groups were treated with AG205(10 μmol/L).At 2 h after OGD/R models were established,cells in the AG205+PROG group were treated with PROG(20 μmol/L).After 24 h of reoxygenation,cell viability was detected using CCK-8,and apoptotic cells were detected us-ing flow cytometry.Results The cell survival rates of the DMSO group and the AG205 group were lower than that of the control group,and that was lower in the AG205 group than in the DMSO group.The cell survival rates of the PROG group and the AG205+PROG group were higher than that of the AG205 group,and that was higher in the PROG group than in the AG205+PROG group(all P<0.05).The apoptosis rates of the DMSO group and the AG205 group were higher than that of the control group,and that was higher in the AG205 group than in the DMSO group;the apoptosis rates of the PROG group and the AG205+PROG group were lower than that of the AG205 group(all P<0.05).Conclusion PROG can inhibit neuronal damage in OGD/R neonatal mice,inhibiting pgrmc1 can reduce OGD/R neuronal activity and in-crease apoptosis,and its mechanism may be related to the regulation of pgrmc1.
胡玉婷;孙小雨;花放
徐州医科大学附属医院康复科,江苏徐州 221006南京医科大学第二附属医院康复科徐州医科大学附属医院神经科
临床医学
黄体酮膜受体组件1pgrmc1信号通路黄体酮氧糖剥夺/复氧细胞模型新生儿缺血缺氧性脑损伤
progesterone receptor membrane component 1pgrmc1 signaling pathwayprogesteroneoxygen glu-cose deprivation/reoxygenation modelneonatal ischemic hypoxic brain injury
《山东医药》 2024 (020)
21-24 / 4
国家自然科学基金项目(81571469);江苏省徐州市重点研发计划项目(KC22130).
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