CAJ | 학술논문

目的观察氯化钆(GdCl3)阻断枯否细胞(KCs)功能对大鼠移植肝脏中KCs核因子(NF)-kB活性的影响,探讨NF-KB活性与FasL及细胞因子基因转录的关系.方法肝移植术后1、2和4 h活杀动物,取肝脏分离KCs,分别用凝胶电迁移率改变分析法(EMSA)、Western blot蛋白印迹法和逆转录-聚合酶链反应(RT-PCR)法检测KCs中NF-KB活性、Fas/FasL基因和蛋白质的表达及细胞因子白细胞介素(IL)-4和TNF-α基因转录.结果 GdCl3组KCs中NF-KB活性受到明显抑制,术后2、4 h FasL和细胞因子IL-4基因转录和蛋白的表达也明显减少(P<0.05),但术后2h TNF-α基因转录没有显著减少(P>0.05).结论 GdCl3可能阻断KCs中NF-kB活性,使KCs中FasL及IL-4的基因和蛋白表达减少,影响移植肝脏免疫耐受的形成.
목적 관찰록화구(GdCl3)조단고부세포(KCs)공능대대서이식간장중KCs핵인자(NF)-kB활성적영향,탐토NF-KB활성여FasL급세포인자기인전록적관계.방법 간이식술후1、2화4 h활살동물,취간장분리KCs,분별용응효전천이솔개변분석법(EMSA)、Western blot단백인적법화역전록-취합매련반응(RT-PCR)법검측KCs중NF-KB활성、Fas/FasL기인화단백질적표체급세포인자백세포개소(IL)-4화TNF-α기인전록.결과 GdCl3조KCs중NF-KB활성수도명현억제,술후2、4 h FasL화세포인자IL-4기인전록화단백적표체야명현감소(P<0.05),단술후2h TNF-α기인전록몰유현저감소(P>0.05).결론 GdCl3가능조단KCs중NF-kB활성,사KCs중FasL급IL-4적기인화단백표체감소,영향이식간장면역내수적형성.
Objective To observe the change of nuclear factor (NF)-kB in Kupffer cells (KCs) ,demonstrate the inhibitory effects of GdCl3 on NF-KB activity and the expression of FasL mRNA and protein,and elucidate the molecular mechanism of liver allograft acceptance. Methods Nuclear pro-tein,membrane protein and total RNA were extracted from KCs in the liver grafts of the rats. NF-kB activi-ty was measured by using electrophoretic mobility shift assay (EMSA). The expression of Fas/FasL mRNA and proteins was detected by using RT-PCR and Western blot, respectively. The expression levels of IL-4 and TNF-α mRNA were examined by using RT-PCR. Results NF-kB activity in the liver transplantation group was higher than that in GdCl3 group 1 h,2 h and 4 h after operation. The expression of FasL and IL-4 mRNA in the GdCl3 group was decreased as compared with that in liver transplantation group both 2 h and 4 h after operation (P < 0.05). The expression of TNF-α mRNA had no significant difference in GdCl3 group both 2 h and 4 h after operation (P > 0.05). Conclusion GdCl3 could suppress the expres-sion of FasL in KCs via inhibiting activity of NF-kB and blocking induction of liver graft tolerance.