CAJ | 학술논문

目的：观察沉默信息调节因子1（SIRT1）对肿瘤坏死因子-α（TNF-α）介导的肠上皮Caco-2细胞屏障功能破坏的影响并探讨其分子机制。方法将Caco-2细胞分3组处理：对照组、TNF-α100 ng/mL 24 h处理组（TNF-α组）及 TNF-α100 ng/mL 24 h处理＋白芦藜醇（Resveratrol）40μm预处理12 h组（TNF-α＋Res组）。分别检测跨上皮电阻（TER）、SIRT1及紧密连接蛋白：ZO-1、occludin的蛋白表达。结果对照组、TNF-α组、TNF-α＋Res组的 SIRT1蛋白相对表达量分别为0．81±0．02、0．43±0．04、0．60±0．03。3组的TER分别为（154．00±5．00）、（97．00±4．00）、（128．00±6．00）Ohm/cm2。TNF-α组 SIRT1的蛋白表达较对照组下降47．00％，TER降低37．00％，经白芦藜醇预处理使TER较TNF-α组升高32．00％。对照组、TNF-α组、TNF-α＋Res组的ZO-1和occludin蛋白相对表达量分别为0．62±0．06和0．57±0．03、0．23±0．05和0．33±0．04、0．41±0．03和0．50±0．02。TNF-α处理后紧密连接蛋白ZO-1、occludin表达显著降低（P＜0．05），而白芦藜醇预处理可缓解该现象，蛋白表达较 TNF-α组分别升高78．00％、51．00％（P＜0．05）。结论 TNF-α处理条件下 SIRT1水平降低，而升高 SIRT1水平可增加肠道紧密连接蛋白 ZO-1、occludin蛋白水平的表达，从而减轻TNF-α对Caco-2细胞构成的上皮屏障功能的损伤。
목적：관찰침묵신식조절인자1（SIRT1）대종류배사인자-α（TNF-α）개도적장상피Caco-2세포병장공능파배적영향병탐토기분자궤제。방법장Caco-2세포분3조처리：대조조、TNF-α100 ng/mL 24 h처리조（TNF-α조）급 TNF-α100 ng/mL 24 h처리＋백호려순（Resveratrol）40μm예처리12 h조（TNF-α＋Res조）。분별검측과상피전조（TER）、SIRT1급긴밀련접단백：ZO-1、occludin적단백표체。결과대조조、TNF-α조、TNF-α＋Res조적 SIRT1단백상대표체량분별위0．81±0．02、0．43±0．04、0．60±0．03。3조적TER분별위（154．00±5．00）、（97．00±4．00）、（128．00±6．00）Ohm/cm2。TNF-α조 SIRT1적단백표체교대조조하강47．00％，TER강저37．00％，경백호려순예처리사TER교TNF-α조승고32．00％。대조조、TNF-α조、TNF-α＋Res조적ZO-1화occludin단백상대표체량분별위0．62±0．06화0．57±0．03、0．23±0．05화0．33±0．04、0．41±0．03화0．50±0．02。TNF-α처리후긴밀련접단백ZO-1、occludin표체현저강저（P＜0．05），이백호려순예처리가완해해현상，단백표체교 TNF-α조분별승고78．00％、51．00％（P＜0．05）。결론 TNF-α처리조건하 SIRT1수평강저，이승고 SIRT1수평가증가장도긴밀련접단백 ZO-1、occludin단백수평적표체，종이감경TNF-α대Caco-2세포구성적상피병장공능적손상。
Objective To observe the influence of silent information regulator factor 1(SIRT1)on TNF-αinduced intestinal epi-thelial Caco-2 cell barrier function destroy and to investigate its molecular machenism.Methods Caco-2 cells were randomly divided into three groups:normal control group (control),TNF-αgroup (TNF-α,100 ng/mL for 24 h)and 100 ng/mL plus 40μm resvera-trol group (TNF-α+Res).Transepithelial electrical resistance (TER)was determined.SIRT1 and the protein expressions of ZO-1 , occludin were examined by using Western blot.Results The relative expression amounts of SIRT1 protein were 0.81 ± 0.02, 0.43±0.04 and 0.60±0.03 respectively.TER of three groups were (154.00±5.00),(97.00±4.00)and(128.00±6.00)Ohm/cm2 respectively.Compared with the control group,the expression of SIRT1 protein was reduced by 47% and TER was decreased by 37.00% in the TNF-αgroup.After resveratrol precondition,TER was increased by 32.00% compared with the TNF-αgroup. The relative expression amounts of ZO-1 and occludin protein in the control group,TNF-αgroup and TNF-α+Res group were (0.62±0.06,0.57±0.03),(0.23±0.05,0.33±0.04)and(0.41±0.03,0.50±0.02)respectively.After TNF-αtreatment,the ex-pressions of ZO-1 and occludin protein were significantly deceased(P<0.05),but the resveratrol precondition could attenuate this phenomenon,compared with TNF-αgroup,the protein expression was increased by 78.00% and 51.00% respectively (P<0.05). Conclusion Under the condition of TNF-αtreatment,the SIRT1 level is decreased,but increasing SIRT1 level could increase the in-testinal tight j unction protein ZO-1 and occludin protein expression,thus alleviate the damage of TNF-αon the epithelial barrier function constituted by Caco-2 cells.