CAJ | 학술논문

目的探讨小檗碱预先给药对缺氧/复氧诱导人肾小管上皮细胞凋亡的影响.方法培养人肾小管上皮细胞(HK-2),以1×106个/ml密度接种于培养皿(2 ml/皿)和96孔培养板(200μl/孔),采用随机数字表法,将其分为4组(n=30):正常对照组(C组)、小檗碱组(B组)、缺氧/复氧组(H/R组)和缺氧/复氧+小檗碱组(H/R+B组).B组和H/R+B组加入10 μmol/L小檗碱孵育2h,随后H/R组和H/R+B组进行缺氧24h复氧3h.测定细胞活力、细胞凋亡率、丙二醛(MDA)、超氧化物歧化酶(SOD)、caspase-3、活化caspase-3、细胞色素c、葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达,并计算Bax表达与Bcl-2表达的比值(Bax/Bcl-2).结果与C组比较,H/R组和H/R+B组细胞活力、SOD活性和caspase-3表达降低,细胞凋亡率、MDA浓度和Bax/Bcl-2升高,活化caspase-3、细胞色素c、GRP78和CHOP表达上调(P＜0.05),B组上述指标差异无统计学意义(P＞0.05)；与H/R组比较,H/R+B组细胞活力、SOD活性和caspase-3表达升高,细胞凋亡率、MDA浓度和Bax /Bcl-2降低,活化caspase-3、细胞色素c、GRP78和CHOP表达下调(P＜0.05).结论小檗碱预先给药可抑制缺氧/复氧诱导人肾小管上皮细胞凋亡,其机制与抑制线粒体应激通路和内质网应激通路有关.
목적 탐토소벽감예선급약대결양/복양유도인신소관상피세포조망적영향.방법 배양인신소관상피세포(HK-2),이1×106개/ml밀도접충우배양명(2 ml/명)화96공배양판(200μl/공),채용수궤수자표법,장기분위4조(n=30):정상대조조(C조)、소벽감조(B조)、결양/복양조(H/R조)화결양/복양+소벽감조(H/R+B조).B조화H/R+B조가입10 μmol/L소벽감부육2h,수후H/R조화H/R+B조진행결양24h복양3h.측정세포활력、세포조망솔、병이철(MDA)、초양화물기화매(SOD)、caspase-3、활화caspase-3、세포색소c、포도당조절단백78(GRP78)화C/EBP동원단백(CHOP)표체,병계산Bax표체여Bcl-2표체적비치(Bax/Bcl-2).결과 여C조비교,H/R조화H/R+B조세포활력、SOD활성화caspase-3표체강저,세포조망솔、MDA농도화Bax/Bcl-2승고,활화caspase-3、세포색소c、GRP78화CHOP표체상조(P＜0.05),B조상술지표차이무통계학의의(P＞0.05)；여H/R조비교,H/R+B조세포활력、SOD활성화caspase-3표체승고,세포조망솔、MDA농도화Bax /Bcl-2강저,활화caspase-3、세포색소c、GRP78화CHOP표체하조(P＜0.05).결론 소벽감예선급약가억제결양/복양유도인신소관상피세포조망,기궤제여억제선립체응격통로화내질망응격통로유관.
Objective To investigate the effect of berberine pretreatment on hypoxia/reoxygenation (H/R)-induced apoptosis in human renal tubular epithelial cells.Methods The human renal tubular epithelial cells were cultured and seeded in culture dishes (2 ml/dish) or 96-well plates (200 μl/well) with the density of 1 ×106/ml.The cells were then randomly divided into 4 groups (n =30 each):normal control group (group C),berberine group (group B),H/R group and H/R + berberine group (H/R + B group).In groups B and H/R + B,berberine 10 μmol/L was added to the culture medium and the cells were incubated for 2 h.Groups H/R and H/R + B were then exposed to 94％ N2-5％ CO2-1％ O2 for 24 h followed by 3 h reoxygenation.The cell viability,apoptotic rate and level of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected.The expression of caspase-3,activated caspase-3,cytochrome c,glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was determined.The ratio of Bax/Bcl-2 was calculated.Results Compared with group C,the cell viability,SOD activity and caspase-3 expression were significantly decreased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were increased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was up-regulated in groups H/R and H/R + B (P ＜ 0.05).Compared with group H/R,the cell viability,SOD activity and caspase-3 expression were significantly increased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were decreased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was down-regulated in group H/R + B (P ＜ 0.05).Conclusion Berberine pretreatment can inhibit H/R-induced apoptosis in human renal tubular epithelial cells,and inhibition of mitochondrial stress pathway and endoplasmic reticulum stress pathway is involved in the mechanism.