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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(P<0.05);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。
OBJECTIVE: To investigate the effect of traditional Chinese medicine of promoting blood circulation and removing blood stasis on the expression of PDGF-BB in the kidney tissue of adriamycin nephropathy rat (AIN) The mechanism of interstitial protection, provide experimental evidence for the early prevention and treatment of tubulointerstitial fibrosis. Methods: 55 SPF male Sprague-Dawley rats weighing 250 ± 20g were randomly divided into 4 groups. 12 normal control group, model control group 13, blood circulation group 15, replenishing qi and Yin group 15. Except the normal group, adriamycin nephropathy model was established by injecting a single dose of ADR 6.5mg / kg into the caudal vein of each group. The normal control group was injected with the same amount of normal saline through the caudal vein. On the day of model start treatment, replenishing qi and activating blood circulation and blood stasis Chinese medicine group twice daily by gastric perfusion Chinese medicine decoction treatment, continuous treatment for 8 weeks. Observed the general situation of experimental rats. At the end of the experiment, blood samples were collected for ALT, ALB, BUN and Scr; The rats were sacrificed at the end of the experiment. The expression of PDGF-BB protein in renal tissue was determined by immunohistochemistry and the expression of PDGF-BB mRNA in rat renal tissue by semi-quantitative RT-PCR. Pathological changes of renal tubules were detected by HE staining. Results: (1) Urine protein excretion peaked at about 8 weeks in adriamycin-induced nephrotic rats; (2) Compared with the model group, each TCM treatment group could slow down the growth of urinary protein excretion; (3) After the experiment, Immunohistochemistry and RT-PCR showed that PDGF-BB protein and mRNA expression were significantly up-regulated compared with the normal control group (P <0.05). Compared with the model control group, the Chinese medicine treatment group could down-regulate PDGF-BB, Huayu group effect is more obvious. (2) The protein and mRNA expression of PDGF-BB in renal tissue, especially in renal tubular epithelial cells of adriamycin-induced nephropathy rats were increased , Consistent with the progression of nephropathy; (3) Chinese medicine for activating blood circulation and removing blood stasis has the effect of slowing the growth of proteinuria, reducing renal pathological damage, especially reducing renal atrophy and interstitial fibrosis, and protecting renal function; 4) Chinese Herbs for Activating Blood Circulation and Removing Blood Stasis can significantly inhibit the expression of PDGF-BB protein and mRNA in renal tissue, and has an early preventive and therapeutic effect on tubulointerstitial fibrosis in rats with adriamycin-induced nephropathy.