丹参多酚酸盐通过抑制内质网应激减轻小肠缺血再灌注损伤的研究  

Salvia polyphenolate reduces the intestinal ischemia-reperfusion injury by inhibiting endoplasmic reticulum stress

作　　者：李贵宾 LI Guibin(Department of Surgery,Handan General,Jizhong Energy Fengfeng Group Hospital,Handan 056002,China)

机构地区：[1]冀中能源峰峰集团有限公司总医院邯郸院区外科,邯郸056002

出　　处：《天津中医药》2021年第9期1195-1200,共6页Tianjin Journal of Traditional Chinese Medicine

基　　金：河北省医学科学研究重点课题计划(20181845)。

摘　　要：[目的]探讨丹参多酚酸盐(SAL)对大鼠小肠缺血再灌注(I/R)后肠组织的保护作用并探索其机制。[方法]将144只大鼠按照随机数字表法平均分为假手术组(Sham组),I/R组,SAL低(10 mg/kg)、中(20 mg/kg)、高(40 mg/kg)剂量组和银杏叶提取物(EGB)组(3.15 mg/kg);造模前30 min腹腔注射给药;采用夹闭肠系膜上动脉60 min的方法复制小肠I/R大鼠模型。再灌注120 min后,采用干湿比重法计算肠组织含水量;分别通过苏木精-伊红(HE)染色法、原位末端转移酶标记(TUNEL)法行肠组织病理学检查和细胞凋亡观察,计算损伤评分(Chiu’s氏评分)和凋亡指数(AI);酶联免疫吸附(ELISA)法检测肠组织炎症因子[C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)]水平;蛋白免疫印迹(Western blot)法检测内质网应激相关蛋白[糖调节蛋白78(GRP78)、c-Jun氨基端激酶(p-JNK)、增强子结合蛋白同源蛋白(CHOP)]、核因子-κB(NF-κB)、激活型半胱氨酸蛋白酶-3(Cleved Caspase-3)蛋白表达。[结果]与I/R组比较,SAL中、高剂量组和EGB组大鼠肠组织含水量降低,差异有统计学意义(P<0.05或P<0.01);肠系膜细胞脱落、固有层分离、炎性细胞浸润等病变和细胞凋亡状况明显改善,Chiu’s氏评分和AI降低,差异有统计学意义(P<0.05或P<0.01);肠组织CRP、TNF-α、IL-1β水平降低,差异有统计学意义(P<0.05或P<0.01);GRP78、p-JNK、CHOP、NF-κB、Cleved Caspase-3蛋白表达下调,差异有统计学意义(P<0.05或P<0.01)。与EGB组比较,SAL高剂量组大鼠Chiu’s氏评分、AI降低,差异有统计学意义(P<0.05或P<0.01);肠组织CRP、TNF-α水平降低,差异有统计学意义(P<0.05或P<0.01);GRP78、CHOP、NF-κB、Cleved Caspase-3蛋白表达下调差异有统计学意义(P<0.05或P<0.01)。[结论]SAL对大鼠小肠I/R损伤具有保护作用;其机制可能与SAL抑制内质网应激通路,进而抑制炎症反应和细胞凋亡有关。[Objective]To investigate the protective effect of Salviaolate(SAL)on intestinal tissue after ischemia/reperfusion(I/R)in rats and explore its mechanism.[Methods]144 rats were randomly divided into sham operation group(Sham group),I/R group,SAL low(10 mg/kg),medium(20 mg/kg),high(40 mg/kg)dose group and Extract of Ginkgo Biloba(EGB)group(positive control group,3.15 mg/kg);the rat models with intestinal I/R injury were made by clipping superior mesenteric artery for 60 mins.30 mins before surgery,the drugs were given by intraperitoneal injection.120mins later,the water content of intestinal tissue was calculated by dry/wet method;the mesentery pathological examination and apoptosis observation were performed by hematoxylin-eosin(HE)staining method and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)method respectively,and the injury score(Chiu’s score)and apoptosis index(AI)were calculated;the levels of inflammatory factors[C reaction protein(CRP),tumor necrosis factor-α(TNF-α),interleuk-1β(IL-1β)]in intestinal tissue were detected by enzyme linked immunosorbent assay(ELISA)method;the expression of endoplasmic reticulumstress-related proteins[glucose regulated protein 78(GRP 78),Jun N-terminal kinases(p-JNK),C/Ebp-homologous protein(CHOP)],nuclear factor kappa-B(NF-κB),Cleved Caspase-3 were detected by Western blot method.[Results]Compared with I/R group,the water content in Salvianolate medium,high dose group and EGB group were significantly decreased(P<0.05 or P<0.01);the mesenteric cell detachment,lamina propria separation,inflammatory cell infiltration and other diseases,and cell apoptosis were significantly improved;the Chiu’s score and AI were significantly reduced(P<0.05 or P<0.01);the level of CRP,TNF-α,IL-1βwere significantly decreased(P<0.01);the expression of RP78,p-JNK,CHOP,NF-κB,Cleved Caspase-3 were significantly down-regulated(P<0.05 or P<0.01).Compared with EGB group,the Chiu’s score and AI were significantly reduced(P<0.05);the level of CRP,TNF-αwe

关 键 词：丹参多酚酸盐 小肠缺血再灌注 内质网应激 炎症 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]