【作者】 王芳；

【导师】 陆付耳；

【作者基本信息】 华中科技大学 ， 中西医结合基础， 2006， 博士

【摘要】 第一部分鱼腥草对1型糖尿病大鼠肾脏早期病变的影响目的为探讨清热解毒法在糖尿病肾病中可能的治疗作用，观察鱼腥草对1型糖尿病大鼠早期及肾功能的影响。方法采用一次性尾静脉注射链脲佐菌素建立Wistar大鼠1型糖尿病模型，随机分为模型组、鱼腥草组、黄芪组、鱼腥草加黄芪组、洛汀新组，另设正常对照组；分别以蒸馏水及四种药物干预8周，检测血糖和血清胰岛素、血脂和24小时尿白蛋白、24小时尿β2微球蛋白、肌酐清除率等的变化。结果与正常对照组相比，模型组及各药物治疗组大鼠体重、胰岛素水平降低，血糖值增高(P＜0.01)；糖尿病模型组大鼠TC、TG、LDL-C升高，HDL-C降低，与正常对照组相比，具有显著性差异(P＜0.05)。鱼腥草、黄芪、鱼腥草加黄芪、洛汀新四组中TC、TG、LDL-C较模型组有所降低，HDL-C升高，具有显著性差异(P＜0.05)，但鱼腥草与其他三种药物干预组之间没有显著性差异(P＞0.05)。与正常对照组相比，24小时尿白蛋白、24小时尿β2微球蛋白、肌酐清除率在模型组中明显增加，具有显著性差异(P＜0.01)；而各治疗组与模型组比较，以上各值降低，具有显著性差异(P＜0.05)，但鱼腥草与其他三种药物干预组之间没有显著性差异(P＞0.05)。与正常对照组相比，模型组及各药物治疗组大鼠肾重/体重比值增加，肾小球面积增加，具有显著性差异(P＜0.05)；各治疗组与模型组比较，肾重/体重比值没有显著性差异，但肾小球面积减小，具有显著性差异(P＜0.05)，各治疗组之间未见显著性差异(P＞0.05)。结论鱼腥草可调节糖尿病大鼠血脂，降低TG、TC、LDL-C水平，升高HDL-C水平；可减少糖尿病大鼠早期肾脏尿白蛋白、β2微球蛋白排泄，降低肌酐清除率，保护肾功能。提示鱼腥草，同黄芪、洛汀新一样，可降低糖尿病肾脏早期高滤过、高灌注压、高通透性，对糖尿病肾脏具有保护功能。第二部分鱼腥草对1型糖尿病大鼠肾脏保护作用的机理探讨目的检测鱼腥草干预后糖尿病大鼠肾脏组织中转化生长因子β1(TGF-β1)、肝细胞生长因子(HGF)、核因子-kappa B(NF-kappa B)等的变化，探讨鱼腥草保护糖尿病大鼠肾脏的机理。方法采用一次性尾静脉注射链脲佐菌素建立Wistar大鼠1型糖尿病模型，随机分为模型组、鱼腥草、黄芪、鱼腥草加黄芪、洛汀新五组，另设正常对照组，治疗8周，应用免疫组化、RT-PCR等方法检测糖尿病大鼠肾脏组织中TGF-β1、HGF、NF-kappa B等的表达。结果糖尿病模型组大鼠肾脏中，TGF-β1、HGF、collagenⅠ及其mRNA、NF-kappa B表达增加，BMP-7及其mRNA表达减少，与正常对照组相比，具有显著性差异(P＜0.05)；各治疗组与之相比，TGF-β1、collagenⅠ及其mRNA、NF-kappa B表达下降，而HGF、BMP-7及其mRNA表达增加(P＜0.05)，同样在各治疗组之间未见显著性差异(P＞0.05)。结论鱼腥草可降低糖尿病大鼠肾脏中炎症调节因子NF-kappa B的表达，降低肾脏组织中TGF-β1、collagenⅠ及其mRNA的表达，增加HGF、BMP-7及其mRNA的表达，提示鱼腥草对肾脏的保护功能可能与其降低糖尿病肾脏进展中促进因子TGF-β1和炎症调节因子NF-kappa B的表达，减少collagenⅠ表达增强HGF、BMP-7的表达有关。第三部分鱼腥草对2型糖尿病大鼠肾脏的影响及机制探讨目的探讨鱼腥草对2型糖尿病大鼠血脂及肾脏功能的影响，并探讨其对肾脏组织中TGF-β1、HGF、BMP-7、NF-kappa B及细胞外基质成分CollagenⅢ表达的影响。方法采用小剂量STZ加高糖高脂高蛋白饮食诱导建立2型糖尿病模型，随机分为鱼腥草、阿司匹林、黄连素、模型组，另设正常对照组；各组动物灌胃治疗8周后，检测血脂、肾功能，并采用免疫组化检测肾脏组织中TGF-β1、HGF、BMP-7、NF-kappa B及细胞外基质成分CollagenⅢ的蛋白表达。结果与正常组比较，模型组大鼠甘油三酯、总胆固醇、游离脂肪酸增高，具有显著性差异(P＜0.05)；与模型组比较，鱼腥草组TC、TG、FFA降低，具有显著性差异(P＜0.05)；鱼腥草组与阿司匹林组、黄连素比较，无显著性差异(P＞0.05)。免疫组化显示，模型组TGF-β1、NF-kappa B、CollagenⅢ、HGF表达增加，BMP-7表达降低，与正常组比较，具有显著性差异(P＜0.05)；鱼腥草组TGF-β1、NF-kappaB、CollagenⅢ表达下降，HGF、BMP-7表达增加，与模型组比较具有显著性差异(P＜0.05)，与阿司匹林、黄连素比较，无显著性差异(P＞0.05)。结论鱼腥草可调节2型糖尿病大鼠，增加胰岛素敏感性，降低24h尿白蛋白、β2微球蛋白及肌酐清除率，保护肾脏功能；鱼腥草可降低2型糖尿病大鼠肾脏组织中TGF-β1、NF-kappa B、CollagenⅢ的表达，增加HGF、BMP-7的表达。鱼腥草对肾脏的保护作用，可能与其抗炎、提供胰岛素敏感性有关，其分子机制可能与鱼腥草降低TGF-β1、NF-kappaB、CollagenⅢ的表达，增加HGF、BMP-7的表达有关。更多还原

【Abstract】 Chapter1 Effects of Houttuynia cordata Thunb on Early Nephropathy in Type 1 Diabetic RatsObjectiveIn order to explore the effect of therapeutic method of clearing-heat and detoxification in traditional Chinese medicine on diabetic renal complication, the study was carried out to investigate the effects of Houttuynia cordata Thunb (HCT) on lipid metabolism and kidney function in type 1 diabetic rats.MethodsMurine type 1 diabetic model was induced by the intravenous injection of streptozotocin and the modeled rats were randomly divided into 5 groups: model group, HCT group, astragalus group, HCT add astragalus group, lotensin group, and normal rats without any treatment were set as normal control group. The modeled rats were treated correspondingly with distilled water, HCT, astragalus, HCT add astragalus, lotensin according their grouping for 8 weeks. Serum TC, TG, HDL-C and LDL-C levels, clearance creatinine, 24h total albumin and 24h β-2 microglobin in urine were determined. ResultsCompared with normal group, murine body weight and insulin level decreased while serum glucose increased significantly in model groups (P<0.01), and furthermore the levels of TC, TG, LDL-C elevated while HDL-C reduced obviously (P<0.05). In comparison with model group, serum levels of TC, TG, LDL-C were decreased (P<0.05) and HDL-C increased in the four treated groups (P<0.05), but there was no statistical difference among these treated groups (P>0.05).It was further demonstrated that albumin in 24h urine and beta 2 microglobin in 24h urine and clearance creatinine increased significantly in model group in comparison with normal group (P<0.05), while the above-mentioned parameters in the four treated groups were remarkably decreased as compared with model group (P<0.05), and there was little difference of those parameters among the four treated groups (P>0.05).Compared with normal group, the glomerular area and the ratio of kidney weight to body weight increased obviously in model group and the other treated groups (P<0.05). Compared with model group, there was no significant difference in K/W ratio among HCT, astragalus, HCT add astragalus, and lotensin groups, but the murine glomerular areas in treated groups decreased significantly (P<0.05), and there was no obviously difference among the treated groups (P>0.05).ConclusionIt is suggested that Houttuynia cordata Thunb might regulate the lipid metabolism in diabetic rats, through which it could decrease the levels of TC, TG and LDL-C and increase HDL-C level. It is also implicated that HCT decoction could protect the kidney function in diabetes rats in view of its reducing the excretion of urine albumin, beta2 microglobin, and clearance rate of creatinine. It could be further inferred that Houttuynia cordata Thunb, like Astragalus membranaceus Bunge and lotensin, might realize its protective effects on renal function through its decreasing the high filtration, high infusion and high permeability of glomeruli in murine early diabetic nephropathy. Chapter 2 Exploration on Protective Effects of Houttuynia Cordata Thunb on Murine Type 1 Diabetic NephropathyObjectiveTo examine the expression of TGF-beta 1, HGF and NF-kappa B and so on in kidney tissue of diabetic rats and to explore the mechanism of Houttuynia cordata Thunb on kidney protection in diabetes rats.MethodsThe type 1 diabetes rat model was induced by the intravenous injection of streptozotocin and the modeled rats were randomly divided into 5 groups and treated with HCT, astragalus, HCT add astragalus, lotensin and saline respectively for 8 weeks. The expression of TGF-beta 1, HGF and NF-kappa B so on in kidney tissue in diabetes rats were determined with imminohistochemistry and RT-PCR.ResultsCompared with normal group, the expression of NF-kappa B, TGF-beta 1, HGF, Collagen I and its mRNA increased significantly, expression of BMP-7 and its mRNA descreased obviously in diabetes model group and the four treatment groups (P<0.05). In comparision with diabetes model group, the expression of NF-kappaB, TGF-beta 1, Collagen I and its mRNA was down-regulated and while the expression of HGF, BMP-7 and its mRNA was up-regulated significantly in the four treatment groups (P <0.05). Regarding the these parameters, there was no obviously difference among the four treatment groups (P >0.05).ConclusionThe result suggested that HCT might protect the kidney tissue of diabetic rats through reducing the expression of NF-kappa B, TGF-beta 1, Collagen I and its mRNA and upregulating the expression of HGF, BMP-7 and its mRNA.Chapter 3 Protective Effect of Houttuynia Cordata Thunb on Murine Type 2 Diabetic Nephropathy and Its MechanismObjectiveTo investigate the effect of Houttuynia cordata Thunb (HCT) on type 2 diabetic nephropathy and the expression of TGF-beta 1, HGF, BMP-7, Collagen III, NF-kappa B in kidney tissue of type 2 diabetic rats.MethodsMurine model of type 2 diabetes was induced by injecting small dose STZ and high lipid and hign glucose forage. The modeled rats were randomly divided into model, HCT, aspirin, berberine groups treated correspondongly by gastrogage for 8 weeks. The murine blood lipids and kidney function were determined. Expression of TGF-beta 1, HGF, BMP-7, Collagen III, NF-kappa B in diabetic kidney were checked.ResultsCompared with normal control group, the serum levels of TC, TG, FFA increased in type 2 diabetes model group, meanwhile total 24h albumin, beta 2 microglobin in urine and clearance creatinine descreased. Compared with model group, the levels of TC, TG, FFA in HCT, aspirin and berberine groups were significantly lowered, but there was no statistical difference among the treatment groups. The expression of TGF-beta 1, NF-kappa B, HGF, Collagen III increased and BMP-7 decreased in type 2 diabetes model group, as compared with normal group. Compared with model group, the expression of TGF-beta 1, NF-kappa B, Collagen III in HCT group decreased while HGF and BMP-7 increased significantly, but among the treatment groups, there was no statistical difference.ConclusionHCT could regulate lipids metabolism and improve insulin sensitivity, protect kidney function. The therapeutic mechanisms might be closely correlated with its improving insulin resistance and up-regulating the kidney expression of HGF, BMP-7 and downregulating the expression of TGF-beta1, Collagen III and NF-kappa B.更多还原