人白细胞介素10基因治疗大鼠重症急性胰腺炎实验研究

目的　观察人白细胞介素 10 (IL 10 )基因治疗大鼠重症急性胰腺炎 (SAP)的疗效。方法　SD大鼠随机分为正常对照组、SAP组、脂质体组、不包含或包含IL 10基因的真核表达质粒pcDNA3与阳离子脂质体复合物组 (n =2 0 )。采用胰腺被膜下均匀注射牛磺胆酸钠建立大鼠SAP模型。SAP诱发 0 5h后腹腔注射给药。采用双抗体夹心ELISA方法检测胰腺、肝脏和肺组织中IL 10浓度 ,并观察血淀粉酶、组织学、组织肿瘤坏死因子 (TNF)及一周死亡率的变化。结果　IL 10基因治疗 2 4h后胰腺、肝脏和肺组织IL 10浓度较SAP组显著升高 (P <0 0 5 ) ;血淀粉酶由 2 0 30 0U/L± 110 0U/L降至 680 0U/L± 70 0U/L(P <0 0 5 ) ;胰腺组织学评分由 4 1± 0 2减轻至 3 2± 0 3(P <0 0 5 ) ;胰腺、肝脏和肺组织TNF浓度显著降低 (P <0 0 5 ) ;一周死亡率由 90 %降至 30 % (P <0 0 5 )。而治疗对照组与SAP组相比无显著性差异。结论　阳离子脂质体介导的pcDNA3 IL 10基因治疗能够显著改善重症急性胰腺炎病情 ,降低死亡率

Therapeutic effects of human interleukin 10 gene transfer on severe acute pancreatitis in rats,an experimental study

OBJECTIVE: To study the therapeutic effects of human interleukin 10 (IL-10) gene transfer on severe acute pancreatitis (SAP) in rats. METHODS: Twenty healthy SD rats were injected intraperitoneally with SA liposome, SA liposome/pcDNA3 or SA liposome/pcDNA3-IL-10. Another twenty SD rats were randomly divided into five groups: rats in one group underwent laparotomy only (normal control), and SAP was induced in the other 4 groups induced by homogeneous injection of sodium taurocholate beneath the pancreatic capsule. Among the 4 SAP groups, one group did not receive any drugs, and liposomes, pcDNA3 or pcDNA3-IL-10 complexed with cationic liposomes were administered to the other groups. Drugs were administered by a single intraperitoneal injection thirty minutes after SAP had been induced. The levels of IL-10 in pancreas, liver and lungs were determined by ELISA kits. The level of serum amylase, histology, and tissue tumor necrosis factor (TNF) were assessed and mortality rate was observed in different groups for one week. RESULTS: The levels of IL-10 in the pancreas, liver and lung 24 hours after IL-10 gene transfer, increased significantly (all > 350 pg/g), and then gradually decreased, however, the levels of IL-10 were still significantly higher that those in the control groups (P < 0.05) 96 hours later and decreased to normal in one week. The levels of IL-10 of transfer control group were not significantly different from those of the normal control group. The levels of IL-10 expression in pancreas, liver and lungs were increased significantly in the gene therapy group, compared with the SAP group. The serum amylase level was (4 300 +/- 700) U/L in normal control group, increased to (20 300 +/- 1 100) U/L 24 hour after SAP induction without a difference between the therapy control group and SAP group, and decreased to (6 800 +/- 700) U/L after IL-10 gene therapy (P < 0.05). The histological score of pancreas was 4.1 +/- 0.2 24 hours after the induction of SAP, and was 3.2 +/- 0.3 in the IL-10 therapy group. The level of TNF in pancreas, liver, and lungs 24 hours after the induction of SAP was significantly higher than that in normal control group (P < 0.05) and was not different from that in therapeutic control group. However, it was decreased markedly in IL-10 therapy group (P < 0.05). No rat in any group died within 2 days after onset. There was no difference of mortality between SAP group and therapeutic control group. The one-week mortality was 90% in the whole SAP group. The one-week mortality of IL-10 gene therapy group was 30 %, significantly lower than that in SAP group (P < 0.05). There was no significant difference in the therapeutic control groups and the SAP group. The values of relative risk of SAP group, SA liposome group, and pcDNA3 group were 12, 8, and 11 times higher than that of gene therapy group (P < 0.05). CONCLUSION: Cationic liposome mediated pcDNA3-IL-10 gene therapy decreases significantly the severity and mortality of SAP.