摘要
目的:观察褪黑素对自身免疫性肝炎大鼠模型外周血淋巴细胞亚群的影响方法:实验于2004-10/2006-10在解放军第一二三医院南京军区肝病中心实验室完成。①实验材料:Wistar大鼠,雄性,3月龄,体质量(230±20)g,购自上海斯莱克实验动物有限责任公司。褪黑素:美国Sigma公司产品,临用前以无水乙醇溶解,再加生理盐水配制,使乙醇浓度为0.1%,置4℃冰箱保存备用。②实验方法:采用弗氏完全佐剂加肝细胞特异性脂蛋白法制作大鼠自身免疫性肝炎模型。将建模成功大鼠随机分为模型对照组、褪黑素注射组及猪促肝细胞生长素注射组,每组20只。褪黑素注射组褪黑素2mg/kg腹腔注射,1次/d,猪促肝细胞生长素注射组2mg/kg猪促肝细胞生长素腹腔注射,1次/d,模型对照组与正常对照组均用含0.01%乙醇的生理盐水腹腔注射。③实验评估:60d后检测各组大鼠外周血淋巴细胞亚群浓度。结果:①CD4+细胞≤39.5%18只,均为褪黑素注射组动物,肝炎组织活动性指数≤8分。22只CD4+细胞>39.5%动物中,4只肝炎组织活动性指数≤8分,其中2只为褪黑素注射组动物,2只为模型对照组动物。18只肝炎组织活动性指数>8分,均为模型对照组动物。②18只CD4+细胞≤39.5%褪黑素注射组动物中,17只肝纤维化指数≤4分,1只肝纤维化指数4分;22只动物CD4+细胞>39.5%,2只肝纤维化指数≤4分,为模型对照组动物,20只肝纤维化指数>4分,褪黑素注射组2只,模型对照组18只。③CD4+细胞≤39.5%,肝组织血管病变均为1级,>39.5%时,2级以上血管病变为90.9%。18只CD4+细胞≤39.5%褪黑素注射动物血管病变为1级。22只CD4+细胞百分比>39.5%,2只血管病变为1级,为模型对照动物,17只血管病变为2级,褪黑素注射组2只,模型对照组15只。3只血管病变为3级,为模型对照动物。④CD4+细胞≤39.5%时,83.3%血管内皮细胞生长因子表达呈弱阳性,CD4+细胞>39.5%时,81.8%血管内皮细胞生长因子表达呈强阳性,提示CD4+细胞与血管内皮细胞生长因子表达有关。⑤模型对照组外周血CD4+细胞数和CD4+/CD8+比值均明显高于其他组(P<0.05),CD8+细胞与其他组无明显差异(P>0.05)。褪黑素注射动物与猪促肝细胞生长素注射动物相比,无明显差异(P>0.05)。结论:褪黑素对自身免疫性肝炎模型大鼠外周血CD4+细胞有较强的抑制作用。
To study the effects of melatonin （MT） on peripheral blood lymphocyte subpopulation of rats with autoimmune hepatitis （AIH）. METHODS： The experiment was performed from October 2004 to October 2006 at Center of Liver Disease of Nanjing Military Area Command of Chinese PLA, the 123 Hospital of Chinese PLA. ①Male Wistar rats aged 3 months with the body mass of （230±20） g were bought from SLAKE experimental animal limited company in shanghai. MT was produced by Sigma in America. MT was dissolved with dehydrated alcohol before utilization and prepared with saline to 0.1% alcohol and stored in refrigerator at 4 ℃. ②The AIH rat models were made by Freund＇s complete adjuvant and liver-specific protein. The rats with AIH were divided randomly into model group, MT group and porcine hepatocyte growth-promoting factors （pHGF） group with 20 in each group. Rats in the MT group received 2 mg/kg MT intraperitoneaUy, once a day. Rats in the pHGF group underwent 2 mg/kg pHGF intraperitoneaUy, once a day. Rats in the model group and normal control group were treated with saline of 0.01% alcohol intraperitoneally, once a day. ③The level of serum peripheral blood lymphocyte subpopulation was detected of each group after 60 days. RESULTS： ①There were 18 rats whose percentages of CD4^＋ cells were lower than or equal to 39.5% and they were all in MT group and their hepatitis histological activity index （HAl） were at most 8 points. In the other 22 rats whose percentages of CD4^＋cells were higher than 39.5%, 4 rats＇ （2 in MT group and 2 in model group） HAl were lower than or equal to 8 points, and 18 rats＇ （all in model group） HAl were higher than 8 points. ②There were 18 rats whose percentages of CD4^＋ cells were lower than or equal to 39.5% and they were all in MT group. Seventeen rats＇ HFI were lower than or equal to 4 points and 1 rat＇s HFI was higher than 4 points. In the other 22 rats whose percentages of CD4^＋cells were higher than 39.5%, 2 rats＇ （all in model group） HFI were lower or equal to 4 points, 20 rats＇ （2 in MT group and 18 in model group） HFI were higher than 4 points. ③Liver vasculopathy were all in Grade 1 in rats whose percentages of CD4^＋cells were lower than or equal to 39.5%. Liver vasculopathy were 90.9% all in Grade 2 in rats whose percentages of CD4^＋ cells were over 39.5%. There were 18 rats whose percentages of CD4^＋ cells were lower than or equal to 39.5% and they were all in MT group and their liver vasculopathy were all in Grade 1. In the other 22 rats whose percentages of CD4^＋ cells were higher than 39.5%, 2 rats＇ （all in model group） liver vasculopathy were in Grade 1, 17 rats＇ （2 in MT group and 15 in model group） liver vasculopathy were in Grade 2, 3 rats＇ （all in model group） liver vasculopathy were in Grade 3. ④83.3% rats＇ expression of vascular endothelial cell growth factor （VEGF） in liver cells was weak positive when the percentages of CD4^＋ cells were lower than or equal to 39.5%. 81.8% rats＇ expression of VEGF in liver cells was strong positive when the percentages of CD4^＋ cells were higher than 39.5%. It suggested that the percentages of CD4^＋ cells in peripheral blood were related with the expression of VEGF in liver cells. ⑤The number of CD4^＋ cells and the proportionality of CD4^＋/CD8^＋ in model group were significantly higher than those in other groups （P 〈 0.05）. There was no significant deviation of the number of CD8^＋ cells between the model group and other groups （P 〉 0.05）. To contrast with those of pHGF group, the number of CD4~ cells, CD8^＋ cells and the proportionality of CD4^＋/CD8^＋ in MT group had no significant deviation （P 〉 0.05）. CONCLUSION： MT can strongly inhibit the CD4^＋ cells of peripheral blood lymphocyte subpopulaUon of AIH rats.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第33期6577-6580,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
作者简介
丁体龙，男．江苏省盐城市人．汉族．1998年第一军医大学毕业，主治医师．主要从事肝病的基础与临床研究。dtLdragon2003@yahoo.com．cn
