中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2015年
4期
493-498,499
,共7页
李永金%杨开勇%张谊%陈月芳%端礼荣%黄晓佳
李永金%楊開勇%張誼%陳月芳%耑禮榮%黃曉佳
리영금%양개용%장의%진월방%단례영%황효가
喹啉酸%PC1 2 细胞%细胞损伤%细胞凋亡%HIF-1 α%信号通路%氧化应激
喹啉痠%PC1 2 細胞%細胞損傷%細胞凋亡%HIF-1 α%信號通路%氧化應激
규람산%PC1 2 세포%세포손상%세포조망%HIF-1 α%신호통로%양화응격
quinolinic acid%PC1 2 cells%cell injury%cell apoptosis%HIF-1 α%signaling pathway%oxidative stress
目的:探讨低氧诱导因子-1(hypoxia inducible factor-1α,HIF-1α)在喹啉酸诱导大鼠肾上腺嗜铬细胞瘤 PC12细胞损伤中的作用。方法用不同浓度的喹啉酸(2.5,5和10 mmol·L -1)处理 PC12细胞24 h 诱导细胞损伤,采用噻唑蓝还原法和乳酸脱氢酶漏出率检测法测定细胞损伤程度,采用丙二醛和超氧化物歧化酶试剂盒检测细胞内氧自由基水平, Hoechst 33258单荧光染色法观察细胞凋亡,免疫荧光染色法检测 HIF-1α在细胞内的表达,免疫印迹法检测细胞 HIF-1α、蛋白激酶 B (Akt)、磷酸化 Akt (phosphorylated-Akt,p-Akt)、淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)和 Bcl-2相关 X 蛋白(Bcl-2 associated X protein,Bax)的表达。结果喹啉酸可剂量依赖性地诱导 PC12细胞损伤,导致细胞内氧自由基增多和细胞凋亡。同时,喹啉酸可使PC12细胞 HIF-1α表达上调并聚集于核内;p-Akt 表达增加, Bax/Bcl-2表达比例增加。结论HIF-1α和 Akt 介导了喹啉酸诱导 PC12的细胞凋亡;此外,氧化应激过程也可能参与了细胞损伤。
目的:探討低氧誘導因子-1(hypoxia inducible factor-1α,HIF-1α)在喹啉痠誘導大鼠腎上腺嗜鉻細胞瘤 PC12細胞損傷中的作用。方法用不同濃度的喹啉痠(2.5,5和10 mmol·L -1)處理 PC12細胞24 h 誘導細胞損傷,採用噻唑藍還原法和乳痠脫氫酶漏齣率檢測法測定細胞損傷程度,採用丙二醛和超氧化物歧化酶試劑盒檢測細胞內氧自由基水平, Hoechst 33258單熒光染色法觀察細胞凋亡,免疫熒光染色法檢測 HIF-1α在細胞內的錶達,免疫印跡法檢測細胞 HIF-1α、蛋白激酶 B (Akt)、燐痠化 Akt (phosphorylated-Akt,p-Akt)、淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)和 Bcl-2相關 X 蛋白(Bcl-2 associated X protein,Bax)的錶達。結果喹啉痠可劑量依賴性地誘導 PC12細胞損傷,導緻細胞內氧自由基增多和細胞凋亡。同時,喹啉痠可使PC12細胞 HIF-1α錶達上調併聚集于覈內;p-Akt 錶達增加, Bax/Bcl-2錶達比例增加。結論HIF-1α和 Akt 介導瞭喹啉痠誘導 PC12的細胞凋亡;此外,氧化應激過程也可能參與瞭細胞損傷。
목적:탐토저양유도인자-1(hypoxia inducible factor-1α,HIF-1α)재규람산유도대서신상선기락세포류 PC12세포손상중적작용。방법용불동농도적규람산(2.5,5화10 mmol·L -1)처리 PC12세포24 h 유도세포손상,채용새서람환원법화유산탈경매루출솔검측법측정세포손상정도,채용병이철화초양화물기화매시제합검측세포내양자유기수평, Hoechst 33258단형광염색법관찰세포조망,면역형광염색법검측 HIF-1α재세포내적표체,면역인적법검측세포 HIF-1α、단백격매 B (Akt)、린산화 Akt (phosphorylated-Akt,p-Akt)、림파류/백혈병-2(B cell lymphoma/lewkmia-2,Bcl-2)화 Bcl-2상관 X 단백(Bcl-2 associated X protein,Bax)적표체。결과규람산가제량의뢰성지유도 PC12세포손상,도치세포내양자유기증다화세포조망。동시,규람산가사PC12세포 HIF-1α표체상조병취집우핵내;p-Akt 표체증가, Bax/Bcl-2표체비례증가。결론HIF-1α화 Akt 개도료규람산유도 PC12적세포조망;차외,양화응격과정야가능삼여료세포손상。
Aim To investigate the role of HIF-1 αin PC1 2 cell injury induced by quinolinic acid.Methods PC1 2 cells were treated with quinolinic acid at the do-ses of 2.5,5 and 1 0 mmol·L -1 ,the cell viability was determined by MTT reduction assay and LDH as-say,the intracellular levels of oxygen species was measured by assessing SOD and MDA levels,cell ap-optosis was determined by Hoechst 33258 staining,the intracellular distribution of HIF-1 αwas examined by HIF-1 αimmunostaining,and the expressions of HIF-1 α,Akt,p-Akt,Bcl-2 and Bax were determined by im-munoblotting analysis.Results Quinolinic acid in-duced cell injury in PC1 2 cells in a dose-dependent manner,and potentiated oxygen radical production and cell apoptosis.In addition,quinolinic acid enhanced HIF-1 αexpression and accumulation in nuclei.The p-Akt expression and Bax/Bcl-2 ratio was increased by quinolinc acid in PC1 2 cells.Conclusions HIF-1 αand Akt mediate qunolinc acid-induced cell apoptosis in PC1 2 cells.And cellular oxidative stress may con-tribute to the injury as well.