摘要
背景:人参皂苷Rg3在脑缺血疾病中具有一定的神经保护作用,但其对氧糖剥夺/复糖复氧损伤的PC12细胞是否也有保护作用,目前尚不明确。目的:研究人参皂苷Rg3对氧糖剥夺/复糖复氧损伤的PC12细胞是否有保护作用以及相关机制。方法:构建高分化PC12细胞的氧糖剥夺/复糖复氧损伤模型,采用MTT、乳酸脱氢酶、Calcein-AM/PI染色、Western blot、流式检测氧糖剥夺/复糖复氧损伤后对PC12细胞的影响,以及不同浓度人参皂苷Rg3处理PC12细胞后的保护作用,并且使用PI3K/AKT通路抑制剂LY294002和AKT激活剂SC79从PI3K/AKT信号通路研究其作用机制。结果与结论:①氧糖剥夺/复糖复氧损伤模型能随着缺糖缺氧时间的延长从而降低PC12细胞的存活率,促进细胞凋亡,上调p-PI3K/PI3K、p-AKT/AKT蛋白比值,促进NLRP3炎性小体、肿瘤坏死因子α、白细胞介素1β、BAX蛋白的表达,促进乳酸脱氢酶的释放;②人参皂苷Rg3在一定浓度范围内能提高氧糖剥夺/复糖复氧损伤的PC12细胞存活率、减少乳酸脱氢酶的释放,抑制细胞凋亡,降低p-PI3K/PI3K和p-AKT/AKT蛋白比值,下调NLRP3炎性小体、白细胞介素1β、肿瘤坏死因子α、BAX蛋白的表达,还会降低活化的caspase-9蛋白表达,促进活化的caspase-3蛋白表达;③LY294002对氧糖剥夺/复糖复氧损伤的PC12细胞的保护作用与人参皂苷Rg3相当,而SC79可以减轻人参皂苷Rg3对PC12细胞的保护效果。因此,人参皂苷Rg3可以通过抑制PI3K/AKT信号通路,抑制PI3K和AKT的磷酸化,发挥抗炎、抗凋亡的作用,减轻氧糖剥夺/复糖复氧对PC12细胞造成的损伤。
BACKGROUND:Ginsenoside Rg3 can protect the brain from ischemia injury.However,it is unclear whether ginsenoside Rg3 can protect PC12 cell against oxygen-glucose deprivation/reoxygenation(OGD/R)injury.OBJECTIVE:To investigate the protective role and underlying mechanism of ginsenoside Rg3 in OGD/R-injured PC12 cells.METHODS:A model of OGD/R-injured PC12 cells was constructed.Protective effects of ginsenoside Rg3 at different concentrations on OGD/R-injured PC12 cells were detected using MTT,lactic dehydrogenase,Calcein-AM/PI cell apoptosis double staining assay,western blot assay,and flow cytometry.PI3K/AKT pathway inhibitor LY294002 and AKT activator SC79 were involved to reveal the potential mechanisms of ginsenoside Rg3.RESULTS AND CONCLUSION:(1)In this study,OGD/R exposure successfully induced cell injury,which reduced cell survival and promoted cell apoptosis in a time-dependent manner,up-regulated p-AKT/AKT and p-PI3K/PI3K ratio,increased the levels of NLRP3 inflammasomes,tumor necrosis factor-α,interleukin-1βand BAX,and promoted the release of lactic dehydrogenase.(2)Ginsenoside Rg3 at a certain range enhanced the viability and reduced apoptosis and lactic dehydrogenase release of OGD/R-injured PC12 cells.Ginsenoside Rg3 down-regulated the ratio of p-AKT/AKT and p-PI3K/PI3K,reduced NLRP3,interleukin-1β,tumor necrosis factor-αand BAX levels.Ginsenoside Rg3 also suppressed the expression of activated caspase-9 and up-regulated the expression of activated caspase-3.(3)LY294002 shared the same protective effect as ginsenoside Rg3 whereas SC79 reversed this outcome.Therefore,ginsenoside Rg3 protects PC12 cells from OGD/R injury mainly through inhibiting the PI3K/AKT pathway and phosphorylation of PI3K and AKT,which may exert an anti-inflammatory and anti-apoptotic effect.
作者
钟京霖
曹惠敏
潘亚茹
简文轩
王奇
Zhong Jinglin;Cao Huimin;Pan Yaru;Jian Wenxuan;Wang Qi(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510000,Guangdong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第2期177-183,共7页
Chinese Journal of Tissue Engineering Research
基金
广东省高等学校重点实验室项目(2019KSYS005),项目负责人:王奇
广东省科技计划国际合作项目(2020A0505100052),项目负责人:王奇。
