rIL-10基因干预对肝纤维化大鼠肝组织中胶原、MMP13及TIMP1表达的影响

摘要

Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.%目的：观察胶原、基质金属蛋白酶13（Matrix metalloproteinase 13，MMP13）及其抑制物基质金属蛋白酶抑制物-1（Tissue inhibitor of metalloproteinase 1，TIMP1）在猪血清诱导的肝纤维化大鼠肝组织中的表达及大鼠白介素10（rat inter-leukin-10， rIL-10）基因干预的影响，探讨rIL-10基因抗肝纤维化的作用机制。方法：30只清洁级SD大鼠随机分为正常对照组（C组）和纤维化模型组。 C组每周腹腔注射2次生理盐水，每次0．5 ml，共8周；纤维化模型组每周腹腔注射2次猪血清，每次0．5 ml，共8周。至造模第5周开始模型组随机分为纤维化组（M组），rIL-10基因干预组（I组）及空质粒对照组（P组）。 C组和M组大鼠尾静脉注射林格氏液作为试剂对照，I组大鼠尾静脉注射rIL-10质粒pcDNA3-rIL-10， P组大鼠尾静脉注射pcD-NA3空质粒，每周1次。于第8周末处死所有大鼠。天狼猩红染色检测各实验组大鼠肝组织胶原沉积情况，S-P免疫组化检测各组大鼠肝脏MMP13及TIMP1表达的情况。结果：天狼猩红染色显示：与C组比较，猪血清诱导的肝纤维化模型M组及空质粒对照P组大鼠肝组织中胶原沉积面积显著增多（P＜0．01）；与M组和P组比较，rIL-10基因干预I组大鼠肝组织中胶原的沉积面积显著减少（ P＜0．01）；免疫组织化学染色结果显示：与C组比较，猪血清诱导的肝纤维化模型M组及空质粒对照P组大鼠肝组织中MMP13及TIMP1表达明显增强（P＜0．01），I组纤维化大鼠肝组织中MMP13表达上调而TIMP1表达显著减少（P＜0．01），与M组和P组比较，I组纤维化大鼠肝组织中MMP13及TIMP1表达显著减少（P＜0．01）。结论：rIL-10基因抑制肝纤维化大鼠肝脏胶原沉积与其抑制基质金属蛋白酶抑制物TIMP1的表达相关。