重组质粒pcDNA3.1/C-sis对大鼠FHF的影响  

The effect of the recombinant plasmid pcDNA3.1/C-sis on FHF of rats

作　　者：蔡鹏程[1] 元文峰 丁浩[1] 

机构地区：[1]南昌大学第二附属医院消化内科,南昌330006 [2]乐安县人民医院内二科,江西抚州344300

出　　处：《重庆医学》2017年第19期2604-2607,2611,共5页Chongqing medicine

基　　金：国家自然科学基金资助项目(81300348);江西省青年科学基金资助项目(20151BAB215006);江西省教育厅科学技术研究项目(GJJ150259);江西省卫生计生委科技计划项目(20165199)

摘　　要：目的探讨重组质粒pcDNA3.1/C-sis导入大鼠对暴发性肝功能衰竭(FHF)的影响。方法利用流体力学的方法将重组质粒pcDNA3.1/C-sis导入大鼠肝脏,48h后用内毒素(LPS)+D-半乳糖胺(D-GalN)诱导大鼠FHF;利用荧光定量PCR及蛋白质印迹法(Western blotting)检测C-sis表达;利用苏木精-伊红(HE)染色及测定半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性检测凋亡;利用Western Blotting检测Bcl-2和Bax的表达变化;计算大鼠24h观察期内的死亡率。结果 FHF+C-sis质粒组的C-sis mRNA和蛋白表达水平较正常对照组和FHF+空载质粒组明显增加,差异均有统计学意义(P<0.01)。与正常对照组相比,FHF+林格液注射组和FHF+空载质粒组肝脏细胞凋亡增多;与FHF+空载质粒组相比,FHF+C-sis质粒组肝脏细胞凋亡减少。与正常对照组相比,FHF+林格液注射组大鼠肝脏组织中Caspase-3的表达增多(P<0.01);与FHF+空载质粒组相比,FHF+C-sis质粒组大鼠肝脏组织中Caspase-3的表达减少(P<0.05)。与正常对照组相比,FHF+林格液注射组大鼠肝脏组织中Bcl-2表达明显减少(P<0.01),Bax表达明显增多(P<0.01);而与FHF+空载质粒组相比,FHF+C-sis质粒组大鼠肝脏组织中Bcl-2表达增多(P<0.05),Bax表达减少(P<0.05)。在24h观察期内,正常对照组大鼠均存活,FHF+林格液注射组及FHF+空载质粒组大鼠死亡率分别为70.0%和80.0%;而FHF+C-sis质粒组大鼠仅20.0%死亡。结论 C-sis基因可减弱LPS+D-GalN诱导的大鼠FHF。Objective To explore the effect of the recombinant plasmid pcDNA3.1/C-sis on fulminant hepatic failure （FHF） in rats. Methods 48 h after recombinant plasmid pcDNA3.1/C-sis being imported into rat liver by using the method of fluid me- chanics,FHF in rats was induced by endotoxin （LPS）＋D-galactosamine （D-GaIN）. With fluorescence quantitative PCR and West- ern blotting,C-sis expression was tested. The apoptosis of rat liver was detected by using HE staining and measuring Caspase-3 ac- tivity. The expression changes of Bcl-2 and Bax were examined through using Western blotting. The mortality rate of rats was cal- culated during 24 h observation period. Results Compared with the normal control group and FHF＋ empty plasmid group, C-sis mRNA and protein expression levels were increased significantly in the FHF＋C-sis plasmid group,there were statistically signifi- cant differences （P〈0. 01）. Compared with the normal control group, the apoptotic hepatocytes were increased in the FHF＋ Ringer＇s solution injection group and FHF＋ empty plasmid group;compared with FHF＋ empty plasmid group, the apoptotic hepa- tocytes in the FHF＋C-sis ptasmid group were decreased. Compared with the normal control group,Caspase-3 expression level was increased in the FHF＋Ringer＇s solution injection group （P〈0.01） ;compared with the FHF＋empty plasmid group,Caspase-3 ex- pression level in the FHF＋C-sis plasmid group was decreased （P〈0.05）. Compared with the normal control group,Bcl-2 expres- sion level was decreased significantly （P〈0.01） ,and Bax expression level was increased significantly （P〈0.01） in the FHF＋ Rin- gerI s solution injection group ; compared with the FHF＋ empty plasmid group, the Bcl-2 expression level was increased （P〈0.05）, and Bax expression was decreased （P〈0.05）in the FHF＋C-sis ptasmid group. During the 24 h observation period,all rats in the normal control group were alive;the mortality rates of the FHF＋ Ringerrs solution injection group

关 键 词：原癌基因蛋白质C-sis 重组质粒 大鼠 肝功能衰竭 急性 

分 类 号：R575[医药卫生—消化系统]