TNF-α a antisense transfer improves ischemia / reperfusion injury of the hepatic graft

移植肝的冷缺血/再灌注损伤，这是肝移植中尚未解决的问题，其原因是激活的枯否细胞释放炎性细胞因子。我们最近设计了一种由日本血凝病毒融合的阳离子脂质体组成的新型载体，即阳离子脂质体(HVJ-CL)，并利用该载体系统评价了异硫氰酸荧光素(FITC)标记的寡脱氧核苷酸(FITC-α)对库普弗细胞的细胞毒性和效率，以及反义肿瘤坏死因子-ODN对鼠肝移植缺血/再灌注损伤的保护作用。移植肝中95%以上的ED2阳性Kupffer细胞在FITC-ODN转移后4小时内识别到FITC信号。在八分之一的载药量和脂类的情况下，L组的转移率(18.5±3.1%)明显高于L组(10.4±1.2%，p&lt；0.01)，而载体诱导的肝细胞胞浆酶的释放在两组间无显著差异。肿瘤坏死因子-α反义转移治疗可显著抑制肝移植后4h肝细胞胞浆酶的释放、血清肿瘤坏死因子-α水平及枯否细胞肿瘤坏死因子-α的免疫组织化学表达。该方法为大鼠移植肝枯否细胞提供了一种安全有效的ODN载体

Cold ischemia/reperfusion injury of the hepatic graft, an unsolved problem in liver transplantation that has been attributed to the release of inflammatory cytokines from activated Kupffer cells. We recently designed a novel vehicle composed of cationic liposomes fused with hemagglutinating virus of Japan (HVJ), namely HVJ-cationic liposomes (HVJ-CL), and using this vector system, we assessed the cytotoxicity, efficiency of Fluorescein isothiocyanate (FITC)-labeled oligodeoxynucleotides (FITC-ODNs) delivery to Kupffer cells and effects of TNF-α antisense delivery against ischemia/reperfusion injury in a rodent hepatic graft. FITC signals were recognized in over 95% of ED2-positive Kupffer cells of transplanted liver 4hrs after FITC-ODNs transfer. The transfer efficiency (18.5 ± 3.1%) using HVJ-CL applied at one-eighth the amount of HVJ and lipids of conventional HVJ-liposomes (HVJ-L) was much higher than that achieved by HVJ-L (10.4 ± 1.2%, p<0.01), while there was no signincant difference in the vector-induced release of cytosolic enzymes from hepatocytes into the blood circulation between HVJ-CL and HVJ-L groups. Treatment with TNF-α antisense transfer significantly suppressed the release of cytosolic enzymes from hepatocytes, the serum levels of TNF-α and immunohistochemical expression of TNF-α by Kupffer cells 4hrs after liver transplantation. This HVJ-CL method provides a safe and effective ODNs delivery to Kupffer cells of hepatic grafts in rats