【摘要】 目的:探讨槐耳菌质、枸杞子和黄精组方(简称槐杞黄)对免疫性肾病的疗效及其机制。方法:雌性BABL/c小鼠随机分为3组,即正常对照组、IgA肾病模型组、槐杞黄治疗组,每组10只。IgA肾病组和槐杞黄治疗组用牛血清白蛋白酸化水灌胃、牛血清白蛋白以及葡萄球菌肠毒素尾静脉注射免疫建立IgA肾病模型,正常对照组以酸化水灌胃,磷酸盐缓冲溶液尾静脉注射作为对照。槐杞黄治疗组于免疫8周后按4g/kg体重给予槐杞黄浸膏干预,正常对照组、IgA肾病模型组以等量的生理盐水作为对照。观察3组小鼠血清白蛋白、尿素、肌酐24h尿蛋白变化以及肾脏病理改变。12周后处死小鼠,无菌分离脾淋巴细胞,并将各组细胞悬液接种于24孔板中置于37℃、50ml/LCO2条件下孵育51h。用ELISA方法检测各组培养细胞上清中细胞因子IL-2、IL-4、INF-γ的水平,比较各组小鼠INF-γ/IL-4的比值。留取血清,用ELISA方法定量检测血清总IgA的浓度,比较各组动物血清IgA水平。结果:1)第八周和第十二周模型组24h尿蛋白均较对照组明显增加(P<0.05),治疗前(8周)槐杞黄治疗组和模型组24h尿蛋白差异无统计学意义(P>0.05),治疗4周后,槐杞黄治疗组24h尿蛋白较模型组明显降低(P<0.05);2)实验结束时模型组血清白蛋白较对照组明显下降(P<0.05),槐杞黄治疗组血清白蛋白较模型组明显升高(P<0.05);尿素氮、肌酐含量三组之间差异无统计学意义;3)病理检查显示对照组无明显异常,模型组出现较显著的肾小球系膜细胞增生和系膜基质增多,毛细血管袢受压,开放欠佳,且免疫荧光显示肾小球系膜区和毛细血管壁有较强的颗粒状或者团块状IgA沉积;槐杞黄治疗组肾小球系膜细胞增生和系膜基质增多情况和IgA沉积程度较模型组明显减轻,毛细血管袢开放良好;4)模型组IL-2、INF-γ的分泌较对照组下降(P<0.01)、IL-4分泌较对照组升高(P<0.01);槐杞黄治疗后的IL-2、INF-γ的分泌较模型组升高(P<0.01),IL-4分泌较模型组下降;5)模型组INF-γ/IL-4的比值较对照组下降(P<0.05),槐杞黄治疗组INF-γ/IL-4的比值较模型组升高(P<0.01),血清总IgA的浓度模型组较对照组升高(P<0.01),槐杞黄治疗组较模型组降低(P<0.05)。结论:槐杞黄治疗可以显著减少小鼠IgA肾病尿蛋白,减轻肾小球系膜增生。其机制可能与其上调IL-2、INF-γ表达以及下调IL-4表达,纠正Thl/Th2失衡的作用密切相关。更多还原

【Abstract】 Objective:Clinical observation on immunity nephritis model mouse treated by Huai Qi Huang and related analysis of mechanism.Methods:Female BABL/c mouse were divided into three groups:normal control group,IgA nephropathy model group and Huai Qi Huang treatment group.The number of every group was equal (n=10).The mouse from model group and Huai Qi Huang treated group were treated with bovine serum albumin by gastric-perfusion, injected with bovine serum albumin and staphylococcal enterotoxin Through the tail wein in order to establish IgA nephropathy model,normal control group were treated with acidified water by gastric-perfusion, injected with phosphate buffer Through the tail wein as control. Huai Qi Huang treated group were treated with Huai Qi Huang (4 g/kg) by gastric-perfusion every day after eight weeks, normal control group and IgA nephropathy group were treated with equal volume of saline by gastric-perfusion every day as control.Twenty-four hours urinary protein excretion,serum biochemical value such as albumin, urea nitrogen, creatinine and pathological changes of kidney.were examined.All mouse were harvested at the 12 th week, The splenic lymphocytes were prepared by aseptic technique.The cell suspention were inoculated in 24-well flat-bottom plates unde37 ℃,50 ml/L CO2, 2.5 μg/L COnA fter 51 h, and the supernatants were harvested for the detection of IL-2、IL-4、INF-γ by ELISA,compare the ratio of INF-γ/ IL-4 among every group. With ELISA method to detect the total IgA concentration in serum among every group.Results:1) The twenty-four hours urinary protein exceretion was higher in model and Huai Qi Huang treatment group than that in control group from 8th week to 12th week(P<0.05), there were no difference between model group and Huai Qi Huang treatment group in urinary protein exceretion befour treament(P>0.05),four weeks after treatment, the twenty-four hours urinary protein exceretion in huai er group was lower than in model group(P<0.05). 2) At the 12 weeks, the serum albumin was significantly lower in model group than in control group(P<0.05),but the serum albumin was significantly higher in huai er group than in model group(P<0.05),there were no difference among three groups in creatinine and urea nitrogen. 3) There is no pathological changes in control group, light microscopy examination showed proliferation of mesangial cells and matrix,partial nephrectomy capillary loops the ball under pressure,open up poor in model group,and immunofluorescence examination showed that srong granular or mass of deposits of IgA were detected in mesangial regions and capillary walls in model group. but the proliferation of mesangial cells and matrix and the IgA deposition was significantly lower in Huai Qi Huang treatment group than in model group,capillary loops well-opening. 4) The secretion of IL-2、INF-γ was significantly lower in model group than in control group(P<0.01),while the secretion of IL-4 was higher in in model group than in control group(P<0.01);but the secretion of IL-2、INF-γ was higher in huaier group than in model group(P<0.01), while the secretion of IL-4 was lower in Huai Qi Huang group than in model group. 5) The ratio of INF-γ/ IL-4 was lower in model group than in control group(P<0.05),but the ratio of INF-γ/IL-4 was higher in Huai Qi Huang group than in model group(P<0.01); Total IgA concentration in serum was higher in model group than in control group(P<0.01),but total IgA concentration in serum was lower in Huai Qi Huang group than in model group(P<0.05).Conclusion:Huai Qi Huang could reduce the urinary protein exceretion relieve the hyperplasia of mesangial cells on immunity IgA nephritis model mouse.This effect is surposed to be closely related to upregulation of INF-γ and IL-2 and downregulation of IL-4 secretion in splenic lymphocytes and then correct the imbalance of TH1/TH2.更多还原