ADR肾病大鼠肾组织中PDGF-BB表达及活血化瘀中药干预的实验研究

目的:通过观察PDGF-BB在阿霉素肾病(AIN)大鼠肾组织的表达及活血化瘀中药对其表达的影响,探讨活血化瘀中药对在阿霉素肾病(AIN)大鼠肾小管间质保护作用的机理,为临床早期防治肾小管间质纤维化提供实验依据。方法:SPF级雄性SD大鼠55只,体重250±20g,随机分为4组。正常对照组12只、模型对照组13只、活血化瘀组15只、补益气阴组15只。除正常组外,其余各组均采用阿霉素(ADR)6.5mg/kg一次性尾静脉注射,建立阿霉素肾病模型,正常对照组予等量生理盐水自尾静脉注射。造模当天开始治疗,补益气阴和活血化瘀中药组每日两次经胃灌服中药汤剂治疗,连续治疗8周。观察实验大鼠的一般情况。造模后第1、2、8周留取尿标本,于实验结束留取血检测谷丙转氨酶(ALT)、白蛋白(ALB)、血尿素氮(BUN)、血肌酐(Scr)水平;在实验结束时宰杀大鼠,取左肾用免疫组化测定大鼠肾组织中PDGF-BB蛋白的表达和RT-PCR半定量法测大鼠肾组织中PDGF-BBmRNA表达;取肾组织以光镜HE染色检测肾小管病理学改变。结果:(1)阿霉素肾病大鼠8周左右尿蛋白排泄达到高峰;(2)各中药治疗组较模型组均能减缓尿蛋白排泄的增长速度;(3)模型对照组在实验结束后免疫组化及RT-PCR观察PDGF-BB蛋白和mRNA表达情况与正常对照组比较明显上调(P0.05);各中药治疗组与模型对照组比较,中药治疗组能下调PDGF-BB,而活血化瘀组效果更明显。结论:(1)实验后2周各造模组大鼠蛋白定量均大于100mg/24h模型制作成功;(2)阿霉素肾病大鼠肾组织尤其肾小管上皮细胞PDGF-BB蛋白和mRNA表达增强,与肾病的进展一致;(3)活血化瘀中药对阿霉素肾病大鼠有减缓蛋白尿增长速度,减轻肾病理损害,尤其减少肾小官萎缩及间质纤维化,保护肾功能;(4)活血化瘀中药还可以明显抑制PDGF-BB蛋白和mRNA在肾组织中的表达,对阿霉素肾病大鼠肾小管间质纤维化具有早期防治作用。

Expression of PDGF-BB in Kidney of Adriamycin Nephropathy Rats and Experiment Research of the Intervetion of Promoting Blood Circulation and Removing Blood Stasis of Traditional Chinese Medicine

Objectives:Through observation of PDGF-BB expression in adriamycin induced nephritis(AIN) rats kidney,the influence of traditional Chinese medicine on its expression was observed.Discussion of promoting blood circulation and removing blood stasis of traditional Chinese medicine in adriamycin induced nephritis(AIN) rats tubulointerstitium protection mechanism for clinical early renal fibrosis was made to provide the evidence.Methods:55 SPF male SD rats,weighted(250±20)g,were randomly divided into 4 groups:the normal control group(12 rats),the model group(13 group),invigorate group(15 rats),relatively group(15 rats).Except the normal group,other groups using adriamycin(6.5mg/kg) to give tail injection.On the day started treatment,Chinese traditional medicine of invigorating qi and Yin was given twice a day,continuously for 8 weeks.Observe the experimental rats generally.After the first 1,2,8 weeks,the blood test alanine aminotransferase(ALT),albumin(ALB),and blood urea nitrogen(BUN) and creatinine(Scr) levels were measured.In the end,rats were killed and left kidney was taken out to test PDGF-BB expression by immunohistochemical determination and RT-PCR half quantitative method was used to test PDGF-BBmRNA expression.HE staining was used to observe the pathology changes.Results:①oxorubicin nephropathy rats at 8 weeks urinary protein achieved excretion peak.②The traditional Chinese medicine treatment groups are able to decrease the growth rate of urinary protein excretion than the model group.3.After the end of the experiment,immunohistochemical observations and PDGF-BB protein expression and mRNA were obviously increased than those in normal control group(P<0.05).Each TCM treatment can decrease PDGF-BB expression compared with model group.Conclusions:①weeks after modeling,rats urinary protein was higher than 100mg/24h which means successful modeling.②PDGF-BB expression in renal tubular epithelial cells of adriamycin nephropathy rats were increased and corresponding to the renal disease developing.③Traditional Chinese medicine of promoting blood circulation and removing blood stasis has effect on reducing kidney damage and protecting kidney.④The drugs also can inhibit PDGF-BB protein and mRNA expressions in kidney tissues of adriamycin nephropathy rats and has prevention effect on tubulointerstitium fibrosis.