白介素-10基因干预对纤维化大鼠TNF-α、PDGF-β和COX-2表达的影响  

Effect of intravenous injection of recombinant IL-10 gene vector on expression of TNF-α, PDGF-β and COX-2 in experimental fibrosis in rats

作　　者：林颖[1] 黄月红[1] 陈治新[1] 王小众[1] 张莉娟[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福建省福州市350001

出　　处：《世界华人消化杂志》2013年第25期2571-2577,共7页World Chinese Journal of Digestology

基　　金：福建省自然基金(青年)资助项目;No.2010J05062;No.2010J01165;福建省卫生厅青年基金资助项目;No.2010-1-102012;福建省临床医学重点专科基金资助项目~~

摘　　要：目的:探讨大鼠白介素-10(rat interleukin-10,rIL-10)基因干预对猪血清诱导的大鼠肝纤维化的拮抗作用及其对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、血小板衍生生长因子-(platelet derivative growth factor-α,PDGF-)和环氧合酶-2(cyclooxygenase-2,COX-2)表达的影响.方法:30只清洁级SD大鼠被随机分为正常对照组(N组)和纤维化模型组,N组每周腹腔注射2次生理盐水,每次0.5 mL,共8 wk;纤维化模型组每周腹腔注射2次猪血清,每次0.5 mL,共8 wk.至造模第5周开始纤维化模型组随机分为纤维化组(M组),rIL-10质粒治疗组(I组),空质粒对照组(P组).N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pcDNA3-rIL-10质粒,P组大鼠尾静脉注射pcDNA3.0空质粒,1次/wk.所有大鼠在第8周末处死,通过HE染色检测各组大鼠肝组织病理改变;SP免疫组织化学检测各组大鼠肝脏TNF-α、PDGF-和COX-2表达的情况.结果:肝组织学病理显示猪血清诱导肝纤维化模型造模成功,rIL-10基因干预能明显降低大鼠肝纤维化的程度,与M组和P组相比,I组纤维化程度明显下降.免疫组织化学结果显示:M组、P组TNF-α、PDGF-和COX-2较N组表达明显增加(0.2206±0.0434,0.2217±0.0518 vs 0.1860±0.0104;0.2891±0.0417,0.2818±0.0272 vs 0.2514±0.0228;0.2174±0.0429,0.2117±0.0221 vs 0.1987±0.0106,均P<0.01);I组TNF-α、PDGF-和COX-2较M组、P组表达显著减低(0.2048±0.0124 vs0.2206±0.0434,0.2217±0.0518;0.2513±0.0165 vs 0.2891±0.0417,0.2818±0.0272;0.1961±0.0142 vs 0.2174±0.0429,0.2117±0.0221,均P<0.01).结论:rIL-10基因干预能有效拮抗猪血清诱导的大鼠肝纤维化形成,其机制可能与其抑制肝组织中TNF-α、PDGF-β和COX-2的表达相关.AIM： To study the effect of intravenous injection of recombinant IL-10 （rIL-10） gene vector on the expression of tumor necrosis factor-α （TNF-α）, platelet derivative growth factor-β （PDGF-β） and cyclooxygenase-2 （COX-2） in pig serum-induced experimental liver fibrosis in rats. METHODS： Thirty SD rats were divided into a normal control and a fibrosis model group. The normal control group was intraperitoneally injected with 0.5 mL of normal sodium twice a week for 8 wk, while the fibrosis model group was injected with equal volume of pig serum for the same duration. At the beginning of the 5th week, the fibrosis model group was further randomly divided into a fibrosis model subgroup, a rIL-10 gene therapy subgroup and an empty vector control subgroup. Rats in the normal control group and fibrosis model subgroup were injected with Ringer’s solution （as a reagent control） via the tail vein weekly, the rIL-10 gene therapy subgroup was injected with rIL-10 plasmid pcDNA3-rIL-10, and the empty vector control subgroup was injected with empty vector pcDNA3. All rats were sacrificed at the end of the 8th week, and liver tissue samples were collected to observe pathological changes in liver tissue by HE staining and to detect the expression of TNF-α, PDGF-β and COX-2 in liver tissue by immunohistochemistry. RESULTS： Histopathology analysis proved that experimental liver fibrosis was induced successfully with pig serum. Compared with the fibrosis model subgroup and empty vector control subgroup, the rIL-10 gene therapy subgroup showed mild liver cell degeneration, decreased inflammatory cell infiltration and collagen deposition. Compared with the normal control group, the expression of TNF-α, PDGF-β and COX-2 was significantly increased in the fibrosis model subgroup and empty vector control subgroup （0.2206 ± 0.0434, 0.2217 ± 0.0518 vs 0.1860 ± 0.0104; 0.2891 ± 0.0417, 0.2818 ± 0.0272 vs 0.2514 ± 0.0228; 0.2174 ± 0.0429, 0.2117 ± 0.0221 vs 0.1987 ± 0.0106, all P 〈 0.01）.

关 键 词：肝纤维化 白介素-10 大鼠 肿瘤坏死因子-α 血小板衍生生长因子-β 环氧化酶-2 

分 类 号：R575.2[医药卫生—消化系统]