CAJ | 학술논문

目的评价缺氧诱导因子-1α(HIF-1α)在七氟醚预处理减轻大鼠皮质神经元凋亡中的作用.方法新生SD大鼠,培养原代皮质神经元,以1×106个/ml的密度接种于6孔板(2 ml/孔),采用随机数字表法,分为4组(n=15):正常对照组(C组)、缺氧复氧组(A/R组)、七氟醚预处理组(SP组)和HIF-1α抑制剂2-甲氧基雌二醇组(H组).采用氧糖缺失90 min,复氧复糖24 h的方法制备皮质神经元缺氧复氧损伤模型;SP组通人2.0％七氟醚2h,随后采用PBS洗涤3次,每次5 min,结束后立即制备缺氧复氧损伤模型;H组加入5 μmol/L2-甲氧基雌二醇孵育72 h时进行七氟醚预处理.采用膜联蛋白V/碘化丙啶双染流式细胞术检测神经元凋亡情况,采用免疫蛋白印迹法测定神经元Bid、Bim、Puma和活化的caspase-3的表达水平.结果与C组比较,A/R组神经元凋亡率升高,Bid、Bim、Puma和活化的caspase-3表达上调(P＜0.05);与A/R组比较,SP组和H组神经元凋亡率降低,SP组Bid、Bim、Puma和活化的caspase-3表达下调(P＜0.05);与SP组比较,H组神经元凋亡率升高,Bid、Bim、Puma和活化的caspase-3表达上调(P＜0.05).结论 HIF-1α介导了七氟醚预处理减轻大鼠神经元凋亡的过程,其机制与下调Bid、Bim和Puma表达有关.
목적 평개결양유도인자-1α(HIF-1α)재칠불미예처리감경대서피질신경원조망중적작용.방법 신생SD대서,배양원대피질신경원,이1×106개/ml적밀도접충우6공판(2 ml/공),채용수궤수자표법,분위4조(n=15):정상대조조(C조)、결양복양조(A/R조)、칠불미예처리조(SP조)화HIF-1α억제제2-갑양기자이순조(H조).채용양당결실90 min,복양복당24 h적방법제비피질신경원결양복양손상모형;SP조통인2.0％칠불미2h,수후채용PBS세조3차,매차5 min,결속후립즉제비결양복양손상모형;H조가입5 μmol/L2-갑양기자이순부육72 h시진행칠불미예처리.채용막련단백V/전화병정쌍염류식세포술검측신경원조망정황,채용면역단백인적법측정신경원Bid、Bim、Puma화활화적caspase-3적표체수평.결과 여C조비교,A/R조신경원조망솔승고,Bid、Bim、Puma화활화적caspase-3표체상조(P＜0.05);여A/R조비교,SP조화H조신경원조망솔강저,SP조Bid、Bim、Puma화활화적caspase-3표체하조(P＜0.05);여SP조비교,H조신경원조망솔승고,Bid、Bim、Puma화활화적caspase-3표체상조(P＜0.05).결론 HIF-1α개도료칠불미예처리감경대서신경원조망적과정,기궤제여하조Bid、Bim화Puma표체유관.
Objective To evaluate the role of hypoxia inducible factor-1α (HIF-1α) in reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning.Methods Primary cortical neurons obtained from neonatal Sprague-Dawley rats were seeded in 6-well plates (2 ml/well),and randomly divided into 4 groups (n =15 each) using a random number table:control group (C group),anoxiareoxygenation (A/R) group,sevoflurane preconditioning group (SP group) and HIF-1α inhibitor 2-methoxyestradiol group (H group).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In group SP,the neurons were exposed to 2.0％ sevoflurane for 2 h followed by 5 min washout for 3 times,and then sevoflurane preconditioning was performed immediately.In group H,sevoflurane preconditioning was performed at 72 h of incubation with 5 μmol/L 2-methoxyestradiol.The apoptosis in neurons was assessed using Annexin V-FITC/PI assay,and apoptosis rate was calculated.The expression of Bid,Bim,Puma and activated caspase-3 in neurons was detected by Western blot.Results Compared with group C,apoptosis rate was significantly increased,and the expression of Bid,Bim,Puma and activated caspase-3 was up-regulated in group A/R.Compared with group A/R,apoptosis rate was significantly decreased,and the expression of Bid,Bim,Puma and activated caspase-3 was down-regulated in group SP.Compared with group SP,apoptosis rate was significantly increased,and the expression of Bid,Bim,Puma and activated caspase-3 was up-regulated in group H.Conclusion HIF-1α mediates reduction of apoptosis in rat neurons by sevoflurane preconditioning,and down-regulated expression of Bid,Bim,and Puma is involved in the mechanism.