摘要
目的探究沉默信息调节因子1(SIRT-1)调控紧密连接蛋白保护肠缺血再灌注后引起的屏障损伤的机制。方法选取2014年7月—2021年7月南京医科大学附属江苏盛泽医院因肠系膜血栓导致的部分小肠坏死行手术治疗的5例患者作为研究对象,采集术中切除坏死肠管及部分正常肠管样本,使用生理盐水冲洗肠内容物后,取其中部分坏死和正常的肠黏膜保存于液氮中。提取黏膜中的蛋白,应用蛋白印迹、免疫荧光、qPCR等方法检测SIRT-1和紧密连接蛋白claudin-4表达情况,检测黏膜屏障电阻(TER)的变化。结果坏死肠管肠黏膜claudin-4表达量为(0.06±0.01),少于正常肠管的(0.22±0.06),差异有统计学意义(t=14.189,P<0.05);SIRT-1激活后,其表达升高,claudin-4表达为(0.61±0.12),高于正常肠管,差异有统计学意义(t=42.334,P<0.05),HE染色可见组织破坏减轻,TER呈现出升高趋势;SIRT-1激活后,TER为(1213.29±46.34)Ω·cm^(2),高于未激活的(992.49±24.65)Ω·cm^(2),差异有统计学意义(t=24.624,P<0.001);SIRT-1抑制后,TER降低为(426.50±33.21)Ω·cm^(2),与未激活比较差异有统计学意义(t=161.956,P<0.001)。结论肠缺血再灌注后多伴随屏障损伤,cladin-4表达量降低,SIRT-1能够对紧密连接蛋白cladin-4起到调控作用,保护屏障功能,为小肠缺血再灌注后损伤的治疗提供新的方向。
Objective To explore the mechanism of silent information regulator 1(SIRT-1)regulating claudin to pro-tect the barrier damage caused by intestinal ischemia-reperfusion.Methods From July 2014 to July 2021,5 patients who underwent surgical treatment for partial intestinal necrosis caused by mesenteric thrombosis in Jiangsu Shengze Hospital Affiliated to Nanjing Medical University were selected as the research objects.During the operation,the ne-crotic bowel and part of the normal bowel were collected.After the intestinal contents were rinsed with normal saline,part of the necrotic and normal intestinal mucosa were taken and preserved in liquid nitrogen.Proteins in the mucosa were extracted,and the expression of SIRT-1 and tight junction protein Claudin-4 was detected by western blot,im-munofluorescence and qPCR,and the changes of mucosal barrier resistance(TER)were detected.Results The ex-pression level of claudin4 in intestinal mucosa of necrotic intestinal tube was(0.06±0.01),which was lower than that of normal intestinal tube(0.22±0.06),and the difference was statistically significant(t=114.189,P<0.05).After the activation of SIRT-1,its expression increased,and the expression of claudin-4 was(0.61±0.12),which was higher than that of normal intestinal tube,and the difference was statistically significant(t=42.334,P<0.05).HE staining showed that tissue damage was reduced,and TER showed an increasing trend.After SIRT-1 activation,the TER was(1213.29±46.34)Ω·cm^(2),which was higher than that of the unactivated(992.49±24.65)Ω·cm^(2),and the dif-ference was statistically significant(t=24.624,P<0.001).After SIRT-1 inhibited treatment,the TER decreased to(426.50±33.21)Ω·cm^(2),and the difference was statistically significant compared with the non-activation(t=161.956,P<0.001).Conclusion After intestinal ischemia-reperfusion,it is often accompanied by barrier damage,and the ex-pression of cladin4 is reduced.SIRT-1 can regulate the tight junction protein cladin-4,protect the barrier function,which provides a n
作者
李晓伟
许华
张传强
LI Xiaowei;XU Hua;ZHANG Chuanqiang(Department of General Surgery,Jiangsu Shengze Hospital Affiliated to Nanjing Medical University,Suzhou,Jiangsu Province,215228 China)
出处
《系统医学》
2022年第13期14-18,共5页
Systems Medicine
基金
南京医科大学康达学院2018年度科研发展基金(SYSD2018044)。
