CAJ | 학술논문

目的观察肝移植排斥反应中移植肝脏内糖皮质激素诱导的肿瘤坏死因子受体配体(GITRL)的表达.方法采用Kamada's二袖套法建立从Lewis到Brown Norway(BN)大鼠的肝移植排斥模型为排斥组(n=5),从BN到BN的肝移植模型为耐受组(n=5).术后24h,抽取血液,取肝脏及分离库普弗(Kupffer)细胞,检测肝脏上GITRL及肿瘤坏死因子(TNF)-α的表达,Kupffer细胞上GITRL的表达,检测血清及细胞上清液中TNF-α的表达.免疫组织化学染色强度采用Image-Pro Plus 6.0图像分析软件分析.结果免疫耐受组和排斥组肝脏内的CITRL的平均染色强度分别为0.113±0.007和0.270±0.018(P＜0.05),TNF-α平均染色强度分别为0.114±0.004和0.141±0.005(P＜0.05),耐受组和排斥组的Kupffer细胞GITRL平均染色强度分别为0.206±0.017和0.337±0.018(P＜0.05),Kupffer细胞的培养上清液中,耐受组和排斥组TNF-α的值分别为(68.66±21.12)、(178.33±29.39)ng/L(P＜0.05).结论在排斥的早期阶段肝脏及Kupffer细胞的GITRL表达增高,监测和干扰GITRL可能有益于肝移植急性排斥反应的早期诊断和处理.
목적 관찰간이식배척반응중이식간장내당피질격소유도적종류배사인자수체배체(GITRL)적표체.방법 채용Kamada's이수투법건립종Lewis도Brown Norway(BN)대서적간이식배척모형위배척조(n=5),종BN도BN적간이식모형위내수조(n=5).술후24h,추취혈액,취간장급분리고보불(Kupffer)세포,검측간장상GITRL급종류배사인자(TNF)-α적표체,Kupffer세포상GITRL적표체,검측혈청급세포상청액중TNF-α적표체.면역조직화학염색강도채용Image-Pro Plus 6.0도상분석연건분석.결과 면역내수조화배척조간장내적CITRL적평균염색강도분별위0.113±0.007화0.270±0.018(P＜0.05),TNF-α평균염색강도분별위0.114±0.004화0.141±0.005(P＜0.05),내수조화배척조적Kupffer세포GITRL평균염색강도분별위0.206±0.017화0.337±0.018(P＜0.05),Kupffer세포적배양상청액중,내수조화배척조TNF-α적치분별위(68.66±21.12)、(178.33±29.39)ng/L(P＜0.05).결론 재배척적조기계단간장급Kupffer세포적GITRL표체증고,감측화간우GITRL가능유익우간이식급성배척반응적조기진단화처리.
Objective To investigate te changes of glucocorticoid induced tumor necrosis factor receptor ligand (GITRL) in hepatic allograft rejection. Methods Liver transplantation from Lewis rats (n = 5 ) to Brown Norway (BN) rats was performed by Kamada' s two-cuff technique as acute rejection group. Liver transplantation from BN to BN rats ( n = 5 ) was performed as tolerance group. Recipients were sacrificed at 24th h postoperation. Blood samples were collected and grafts were harvested, then Kupffer cells were isolated. GITRL and tumor necrosis factor (TNF)-α protein expression in the hver was tested by immunohistochemistry, and the GITRL expression in Kupffer cells by immunocytochemistry. Enzyme linked immunosorbent assay (ELISA) was employed to detect the changes of TNF-α protein in the serum and supernatant. The staining intensity was analyzed by Image-Pro Plus 6. 0 image analysis software. Results At 24th h postoperation, the liver GITRL expression levels in tolerance and rejection groups were 0. 113 ± 0. 007 and 0. 270 ±0. 018, respectively (P ＜0. 05). The TNF-α expression levels in the liver in tolerance and rejection groups were 0. 114 ± 0. 004 and 0. 141 ± 0. 005 respectively ( P ＜ 0.05 ). The GITRL expression levels in Kupffer cells in tolerance and rejection groups were 0. 206 ±0. 017 and 0. 337 ±0. 018 respectively (P ＜0. 05 ). As compared with tolerance group (68. 66 ±21.12) ng/L, TNF-α protein expression levels were up-regulated in the supernatant of rejection group ( 178.33 ± 29. 39 ) ng/L ( P ＜ 0. 05 ).Conclusion The expression of GITRL in the liver and Kupffer cells was increased in the early stage of rejection, and monitoring and interfering GITRL may be useful for the early diagnosis and management of an acute rejection in liver transplantation.