摘要
目的研究大鼠肝移植缺血再灌注后 Kupffer 细胞 CD14基因及蛋白的表达,探讨其在再灌注损伤中的作用。方法分离培养大鼠肝移植缺血再灌注后0(对照组)、2、6、12h(IR 组)的Kupffer 细胞,用逆转录聚合酶联反应(RT-PCR)检测 Kupffer 细胞 CD14 mRNA 的表达,用免疫印迹检测 CD14蛋白合成,用酶联免疫吸附试验(ELISA)法测定培养上清 TNF-α的分泌量。然后在上述时间点的细胞培养液中加入抗 CD14抗体(anti-CD14组),观察 CD14抗体对 TNF-α分泌的影响。结果再灌注后 Kupffer 细胞 CD14 mRNA、蛋白以及 TNF-α随观察时间点呈逐步上升趋势(与对照组相比,P<0.01)。应用抗 CD14抗体后,TNF-α表达较 IR 组明显降低(P<0.01)。结论再灌注后Kupffer 细胞 CD14基因及蛋白的表达明显升高,TNF-α的合成和分泌也明显增强;抗 CD14单抗能明显抑制了 NF-α的产生;CD14在介导 Kupffer 细胞激活和肝移植缺血再灌注损伤中可能起重要作用。
Objective To study the expression of lipopolysaccharide receptor CD14 gene and protein in Kupffer cells and its role in ischemia-reperfusion injury （IRI） of rat liver graft. Methods The Kupffer cells were isolated at 0 （control group）, 2, 6, 12 h （IR group） following IRI. The mRNA expression of CD14 was detected by RT-PCR, protein synthesis of CD14 determined by Western blotting and TNF-α level in the supematant measured by ELISA. Then the Kupffer cells were incubated with anti-CD14 polyclonal antibody and the TNF-α level was determined again. Results After IRI, the expression level of CD14 and TNF-α level were significantly increased in the IR group as compared with the control group （P〈0.01）. Then they were markedly decreased after treatment with anti-CD14 antibody in the IR group as compared with the control group （P〈0.01）. Conclusions LPS following IRI can up-regulate CD14 gene and protein expression in Kupffer cells and synthesis of TNF-α. Anti-CD14 antibody can inhibit the production of TNF-α induced by LPS. CD14 might play an important role in Kupffer cell activation and IRI.
出处
《中华肝胆外科杂志》
CAS
CSCD
2006年第4期264-266,共3页
Chinese Journal of Hepatobiliary Surgery
基金
国家自然科学基金(No.30300337)
关键词
肝移植
缺血再灌注损伤
CD14
KUPFFER细胞
Liver transplantation
Ischemia-reperfusion injury
Lipopolysaccharide receptor CD14
Kupffer cell
