CAJ | 학술논문

目的探讨京尼平减轻棕榈酸对HepG2细胞毒性的作用及机制.方法将HepG2细胞分为4组,分别用牛血清白蛋白、棕榈酸(1 mmol/L)、京尼平(20 μmol/L)或京尼平(20 μmol/L)预处理30 min后棕榈酸(1 mmol/L)孵育24 h,检测细胞活力及乳酸脱氧酶(LDH)释放;孵育16 h后经流式细胞术和Hoechst染色检测细胞凋亡;孵育6 h后实时荧光定量PCR检测糖调节蛋白(GRP)78、CCAAT增强子结合蛋白同源蛋白(CHOP)基因表达,PCR结合电泳检测X盒结合蛋白(XBP)-1剪接体.结果和空白组(牛血清白蛋白)相比,棕榈酸可降低HepG2细胞活力(P＜0.05)、增加LDH释放(P＜0.01)和HepG2细胞凋亡(P＜0.05),上调GRP78、CHOP的表达(P＜0.001)及XBP-1的剪接;和棕榈酸组相比,京尼平增加细胞活力(P＜0.05)、减少LDH释放(P＜0.05),显著抑制细胞凋亡(P＜0.01),降低GRP78、CHOP基因(P＜0.01)的表达.PCR产物电泳显示京尼平减少了XBP-1的剪接.结论京尼平对棕榈酸诱导的细胞凋亡有保护作用,其机制可能与抑制内质网应激有关.
목적 탐토경니평감경종려산대HepG2세포독성적작용급궤제.방법 장HepG2세포분위4조,분별용우혈청백단백、종려산(1 mmol/L)、경니평(20 μmol/L)혹경니평(20 μmol/L)예처리30 min후종려산(1 mmol/L)부육24 h,검측세포활력급유산탈양매(LDH)석방;부육16 h후경류식세포술화Hoechst염색검측세포조망;부육6 h후실시형광정량PCR검측당조절단백(GRP)78、CCAAT증강자결합단백동원단백(CHOP)기인표체,PCR결합전영검측X합결합단백(XBP)-1전접체.결과 화공백조(우혈청백단백)상비,종려산가강저HepG2세포활력(P＜0.05)、증가LDH석방(P＜0.01)화HepG2세포조망(P＜0.05),상조GRP78、CHOP적표체(P＜0.001)급XBP-1적전접;화종려산조상비,경니평증가세포활력(P＜0.05)、감소LDH석방(P＜0.05),현저억제세포조망(P＜0.01),강저GRP78、CHOP기인(P＜0.01)적표체.PCR산물전영현시경니평감소료XBP-1적전접.결론 경니평대종려산유도적세포조망유보호작용,기궤제가능여억제내질망응격유관.
Objective To examine the protective effect of genipin from palmitate-induced cytotoxicity in HepG2 cells and investigate the underlying mechanism. Methods HepG2 cells were divided into 4groups and were treated respectively with bovine serum albumin (BSA) ,palmitate (1 mmol/L) ,genipin(20μmol/L) or palmitate for 24 h after genipin pretreatment for 30 min. Assayed the cell viability and lactate dehydrogenase enzyme (LDH) release. Flowcytometry and Hoechst staining were employed for determination of cell apoptosis after 16 h-treatment. Glucose-regulated protein( GRP)78 and CCAAT enhancer binding protein-homologous protein(CHOP) mRNA expression was quantified by real time PCR while X-box binding protein( XBP)-1 splicing was showed by PCR and electrophoresis after 6 h-treatment. Results Compared with BSA, palmitate decreased cell viability ( P ＜ 0. 05 )while increased LDH release ( P ＜ 0. 01 ). It also significantly induced apoptosis of HepG2 cell (P ＜0.05 ). Expression of GRP78 and CHOP mRNA was up-regulated by palmitate (P＜0.001), so was XBP-1 splicing. Compared with palmitate, genipin pretreatment increased cell viability (P＜0.05), reduced LDH release (P＜0.05) and inhibited apoptosis ( P ＜ 0. 01 ) of HepG2 cell. The expression of GRP78 and CHOP mRNA was decreased by genipin( P ＜ 0. 01 ). Electrophoresis of XBP-1 PCR products showed less spliced XBP-1 in genipin-palmitate treated cells than palmitate treated ones. Conclusion Genipin protects HepG2 cells from palmitate-induced cell apoptosis, which may be mediated by inhibition of endoplasmic reticulum stress.