吡咯二硫氨基甲酸对移植肾细胞毒作用的影响

目的探讨核因子-κB(NF-кB)抑制剂-吡咯二硫氨基甲酸(PDTC),对大鼠移植肾急性排斥反应(AR)中细胞毒作用的影响。方法以SD大鼠和Wistar大鼠作为供鼠,Wistar大鼠作为受鼠,建立同系和同种大鼠原位肾移植模型。实验共分三组,对照组取同系移植受鼠,AR组和PDTC组取同种移植受鼠,其中PDTC组受鼠于术后腹腔注射PDTC 100 mg.kg-1.d-1,其余两组注射等量生理盐水。术后1、3、5和7 d,取各组血清和移植肾组织,检测血清肌酐(Cr)和尿素氮(BUN)水平;观察肾脏组织学改变;real time PCR检测移植肾穿孔素(PFP)、颗粒酶B(GzmB)及FasL mRNA的表达;Western blot检测移植肾NF-кB p65、PFP、GzmB及FasL蛋白的表达。结果 PDTC组大鼠肾功能及肾组织病理损伤均较AR组显著减轻。术后各时间点,AR组和PDTC组PFP、GzmB和FasL mRNA表达均明显高于对照组(P<0.01),PDTC组各mRNA表达较AR组显著降低(P<0.01)。AR组和PDTC组NF-кB P65、PFP、GzmB及FasL蛋白的表达均较对照组显著增高(P<0.01),PDTC组各蛋白表达较AR组明显降低(P<0.05,P<0.01)。结论 PDTC通过抑制NF-кB活化,进而减少PFP、GzmB和FasL的表达,减轻移植肾AR中的细胞毒损伤作用。

Effect of Pyrrolidine Dithiocarbamat on Cytotoxicity in Renal Transplant

Objective To investigate the effect of NF-κB inhibitor-pyrrolidine dithiocarbamat （PDTC） on cytotoxicity of acute rejec- tion in renal transplant. Methods With male SD rats and Wistar rats as the donors and Wistar rats as the recipients as well, syngeneic and allogeneic orthotopic renal transplantation models were established; 3 groups were involved in the experiment： control group （synge- neic, normal saline, i.p.）, AR group （allogeneic, normal saline, i.p.） and PDTC group （allogeneic, NF-r.B inhibitor-pyrrolidine dithiocar- bamat （PDTC） i.p.）; the serum samples and tissues of the transplanted kidney were collected at the time points of ld, 3d, 5d and 7d after transplantation and the levels of Cr and BUN were detected; the histological changes of the kidney were observed; real time PCR detec- tion was made to the expression ofperforin （PFP）, granzymeB （GzmB） and FasL mRNA; the expression ofNF-κBp65, PFP, GzmB and FasL protein were detected by Western blot. Results The renal function in PDTC group was better than that in AR group and the patho- logical injuries were much more alleviated; at each time point after transplantation, the levels of PFP, GzmB and FasL mRNA in AR group and PDTC group were much higher than those in control group （P〈0.01） while the expressions of mRNA in PDTC group de- creased much more than those in AR group （P〈0.01）; the expressions of NF-κB P65, PFP, GzmB and FasL protein in AR group and PDTC group were increased much more than those in control group （P〈0.01）, and the expressions of proteins in PDTC group decreased obviously more than those in AR group （P〈0.05, P〈0.01）. Conclusions PDTC relieves the cytotoxicity in renal transplant of acute rejec- tion by down-regulating the expressions of PFP, GzmB and FasL through inhibiting NF-κB.