摘要
目的观察创伤失血性休克(THS)大鼠肠系膜淋巴液对血管内皮细胞程序性坏死的影响。方法建立THS大鼠模型,收集THS手术(T/HS)及假手术(T/SS)大鼠肠系膜淋巴液;将传代的人脐静脉内皮细胞(HUVECs)分为3组,T/HS、T/SS组分别加入1∶1稀释的T/HS、T/SS大鼠肠系膜淋巴液,对照组加PBS;培养3 h后,MTT法检测细胞活性,Western blot法检测程序性坏死的特异性蛋白受体交互作用蛋白1(RIP1)、RIP3。将HUVECS分为6组,A(D)、B(E)、C(F)组分别加入PBS、细胞程序性坏死阻滞剂Nec-1、溶媒DMSO预处理1 h,D、E、F组分别加入1∶1稀释的T/HS大鼠肠淋巴液。孵育3 h后,观察细胞活性及细胞程序性坏死情况。结果与T/SS组、对照组比较,T/HS组细胞活性降低、RIP1及RIP3表达量增加(P均<0.01);与D、F组比较,E组细胞活性增加、RIP1和RIP3表达量降低(P均<0.01);与A、B、C组比较,E组细胞活性降低、RIP1和RIP3表达量增加(P均<0.01)。结论 THS肠淋巴液能够导致血管内皮细胞程序性坏死,该作用能被Nec-1部分抑制。
Objective To observe the influence of mesenteric lymph on the vascular endothelial cell necmptosis of rats with trauma-hemorrhagic shock (THS).Methods We constructed THS mt models and collected mesenteric lymph from trauma-hemorrhagic shock (T/HS) rats and trauma-sham shock (T/SS) rats.The subcultured human umbilical vein endothelial cells(HUVECs) were divided into 3 groups.T/HS group was cultivated with T/HS mesenteric lymph and T/SS group was cultivated with T/SS mesenteric lymph (all mesenteric lymph were diluted 1∶1 with PBS),and the control group was cultivated with the same amount PBS.After being cultured for 3 h,the cell viability was measured by MTT,and receptor interaction protein 1 (RIP1) and RIP3 were analyzed by Western blot.The subcultured HUVECs were divided into 6 groups,PBS,Nec-1,solvent DMSO were respectively added into A (D),B (E),C (F)groups,1 hour later,T/HS mesenteric lymph that diluted 1 ∶ 1 with PBS was added into D,E and F groups.After 3 h incubation,the cell activity and necroptosis was observed.Results The cell viability was lower,and the expression levels of RIP1 and RLP3 were increased in T/HS group as compared with those of the control group and T/SS group (all P ≤ 0.01).The cell viability was higher,and the expression levels of RIP1 and RIP3 were decreased in the E group as compared with those of D and F groups (all P ≤0.01),but they were respectively lower and increased as compared with A,B and C groups (all P ≤0.01).Condusion Mesenteric lymph of rats with THS can induce vascular endothelial cell necroptosis,and Nec-1 can inhibit the occurrence of necroptosis partially.
出处
《山东医药》
CAS
2014年第8期5-8,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81301624)
国家临床重点专科建设项目(2011-873)