乌司他丁预处理可减轻氧糖剥夺诱导的GES-1细胞损伤
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摘要
目的:观察乌司他丁(UTI)预处理对氧糖剥夺(OGD)诱导人胃黏膜上皮细胞株(GES-1)细胞损伤的影响。方法:采用GES-1细胞株进行实验,设置正常对照组(N组)、氧糖剥夺组(O组)、乌司他丁预处理组(U组)。N组不做任何处理,O组和U组通过三气培养箱和无糖培养基共同作用造成细胞氧糖剥夺损伤,U组在损伤前12 h预先加入乌司他丁处理。损伤处理6 h后,采用CCK-8法测定细胞活力,流式细胞术检测细胞凋亡率,免疫印迹法检测Caspase-3和Cleaved Caspase-3的蛋白表达水平,实时荧光定量PCR法检测细胞内瞬时受体电位香草酸亚型-1(TRPV1)分子m RNA的表达水平。结果:与N组比较,O组细胞存活率显著降低,细胞凋亡率明显升高,Caspase-3和Cleaved Caspase-3表达增加,TRPV1 m RNA表达降低(P<0.05);乌司他丁预处理能显著抑制上述O组的变化,差异有统计学意义。结论:乌司他丁预处理可减轻OGD诱导的GES-1细胞损伤,其机制可能与抑制细胞凋亡以及调控TRPV1的表达有关。
Objective To observe the effects of the preconditioning of ulinastatin on GES-1 cell injuryinduced by oxygen and glucose deprivation(OGD). Methods GES-1 cells were cultured in vitro and divided intothree groups: normal control group(group N), oxygen and glucose deprivation group(group O), and ulinastatinpreconditioning group(group U). The OGD model of GES-1 cells were established by glucose-free medium and three-gas incubator for 6h. Ulinastatin was added to group U 12 h before the deprivation of oxygen and glucose. The cellviability and apoptosis were determined by cck-8 and flow cytometry respectively. Western Blot was used toexamine the protein expression of Caspase-3 and Cleaved Caspase-3. The TRPV1 m RNA expression was measuredby quantitative real-time PCR. Results As compared with group N, the viability of GES-1 was decreased, theapoptotic rate and the expression of Caspase-3 and Cleaved Caspase-3 were increased, and the TRPV1 m RNAexpression decreased greatly in group O(P < 0.05). As compared with group O, the aforementioned changes weresignificantly inhibited in group U. Conclusions Ulinastatin preconditioning could effectively inhibit GES-1 cellinjury induced by OGD, which may be related to the inhibition of apoptosis and the upregulation of TRPV1 m RNAexpression.
Objective To observe the effects of the preconditioning of ulinastatin on GES-1 cell injuryinduced by oxygen and glucose deprivation(OGD). Methods GES-1 cells were cultured in vitro and divided intothree groups: normal control group(group N), oxygen and glucose deprivation group(group O), and ulinastatinpreconditioning group(group U). The OGD model of GES-1 cells were established by glucose-free medium and three-gas incubator for 6h. Ulinastatin was added to group U 12 h before the deprivation of oxygen and glucose. The cellviability and apoptosis were determined by cck-8 and flow cytometry respectively. Western Blot was used toexamine the protein expression of Caspase-3 and Cleaved Caspase-3. The TRPV1 m RNA expression was measuredby quantitative real-time PCR. Results As compared with group N, the viability of GES-1 was decreased, theapoptotic rate and the expression of Caspase-3 and Cleaved Caspase-3 were increased, and the TRPV1 m RNAexpression decreased greatly in group O(P < 0.05). As compared with group O, the aforementioned changes weresignificantly inhibited in group U. Conclusions Ulinastatin preconditioning could effectively inhibit GES-1 cellinjury induced by OGD, which may be related to the inhibition of apoptosis and the upregulation of TRPV1 m RNAexpression.
引文
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[3]MONNIG A A,PRITTIE J E.A review of stress-related mucosal disease[J].J Vet Emerg Crit Care(San Antonio),2011,21(5):484-495.
[4]ZHENG K,SHENG Z,LI Y,et al.Salidroside inhibits oxygen glucose deprivation(OGD)/re-oxygenation-induced H9c2 cell necrosis through activating of Akt-Nrf2 signaling[J].Biochem Biophys Res Commun,2014,451(1):79-85.
[5]DARSHIT B S,RAMANATHAN M.Activation of AKT1/GSK-3beta/beta-Catenin-TRIM11/Survivin pathway by novel GSK-3beta inhibitor promotes neuron cell survival:study in differentiated SH-SY5Y cells in OGD model[J].Mol Neurobiol,2016,53(10):6716-6729.
[6]XU C E,ZHANG M Y,ZOU C W,et al.Evaluation of the pharmacological function of ulinastatin in experimental animals[J].Molecules,2012,17(8):9070-9080.
[7]QIAN Z Y,YANG M F,ZUO K Q,et al.Protective effects of ulinastatin on intestinal injury during the perioperative period of acute superior mesenteric artery ischemia[J].Eur Rev Med Pharmacol Sci,2014,18(23):3726-3732.
[8]HU X T,DING C,ZHOU N.et al.Quercetin protects gastric epithelial cell from oxidative damage in vitro and in vivo[J].Eur J Pharmacol,2015,754(1):115-124.
[9]GUO C,LIANG F L,MASOOD W S,et al.Hydrogen sulfide protected gastric epithelial cell from ischemia/reperfusion injury by Keap1 s-sulfhydration,MAPK dependent anti-apoptosis and NF-kappa B dependent anti-inflammation pathway[J].Eur JPharmacol,2014,725:70-78.
[10]LENG Y X,YANG S G,SONG Y H,et al.Ulinastatin for acute lung injury and acute respiratory distress syndrome:Asystematic review and meta-analysis[J].World J Crit Care Med,2014,3(1):34-41.
[11]HOLZER P.TRP channels in the digestive system[J].Curr Pharm Biotechnol,2011,12(1):24-34.
[12]史孝敏,李泽培.TRPV1在消化系统疾病中的研究进展[J].胃肠病学,2013,18(7):444-446.
[13]EVANGELISTA S.Capsaicin receptor as target of calcitonin gene-ralated peptide in the gut[J].Prog Drug Res,2014,68(68):259-276.
[14]LIU Z B,FEI S J,ZHU S P,et al.Protection of ghrelin postconditioning on hypoxia/reoxygenation in gastric epithelial cells[J].World J Gastroenterol,2012,18(38):5377-5388.
[15]温君琳,屠伟峰,郄文斌,等.舒芬太尼预先给药对浸水束缚应激大鼠下丘脑、胃黏膜TRPV1 m RNA的影响[J].实用医学杂志,2014,30(12):1856-1859.
[16]刘晓毅,车向明.乌司他丁通过抑制肠黏膜细胞凋亡减轻大鼠肠道缺血-再灌注损伤[J].西安交通大学学报(医学版),2010,31(5):566-569.