【作者】 李显波；

【导师】 黄春林；

【作者基本信息】 广州中医药大学 ， 中医内科学， 2009， 博士

【摘要】 1.目的意义本研究主要是对尿毒灵的临床疗效进行观察,对于严格按照诊断标准、纳入标准的情况下选取患者用药,并观察其证候积分、生化等指标变化;同时对于尿毒灵的作用机制,以TGF/smad转导通路为研究指标,进行探索性实验研究;至在寻找DN新的治疗药物,对名老专家经验方进行临床疗效及作用机制同时研究,为开发新药做好准备。2.方法住院和门诊的糖尿病人中筛选出符合DN诊断标准的病人,符合诊断标准,血肌酐178≤(Cr)≤442ummol/l,排除其它原因引起的肾功能不全,观察30例患者,观察三个月,观察用药前后患者症候积分、生化、血脂、肝功、血常规变化情况,及最后疗效。SPF级雄性Wister大鼠48只,体重180-220g,适应性喂养一周。按体重随机分为正常对照组12只,剩余36只用于造模。造模方法选为STZ(链脲佐菌素)+腺嘌呤灌胃:将STZ用缓冲液(2.1g柠檬酸溶于100ml双蒸水中配成A液,2.44g柠檬酸钠溶于100ml双蒸水中为B液,A、B液混合即为缓冲液)配置,36只大鼠禁食12小时,STZ以剂量为60mg/kg体重,一次性腹腔注射,三天后测随机血糖,血糖大于16.1mmol/l的大鼠,造模成功,为减少死亡率血糖大于25mmol/l的大鼠皮下注射普通胰岛素1-2iu,血糖不足16.1mmol/l的大鼠,补注射STZ 20mg/kg;全部成模后,予2%腺嘌呤,300mg/kg灌胃,每7天取一次眶静脉血直至成模。造模过程中死亡5只,成模后剩余31只大鼠,按随机表随机分组,分为模型组、尿毒灵组、尿毒清组;模型组灌胃给予2-4ml生理盐水,尿毒灵组予尿毒灵组0.432g/kg,尿毒清组予尿毒清0.27g/kg;给药两个月后,眶静脉丛取血测生化指标,取部分肾脏做免疫组化,部分肾脏做RT-PCR。3.结果实验研究部分:给药组与模型组相比较,尿毒灵组血肌酐、尿素氮下降有统计学意义;尿毒灵组与模型组相比胆固醇、甘油三酯、低密度脂蛋白胆固醇差异不显著,无统计学意义;TGF-β1、Smad3、Smad4、PAI-1在模型组的表达均较空白组有明显增加,有统计学意义,P＜0.05,尿毒灵组上述蛋白的表达均较模型组有明显下降;RT-PCR检测大鼠肾组织TGF-β1mRNA、Smad3 mRNA、Smad4 mRNA,荧光定量PCR检测各指标在肾组织的表达,模型组与空白组比较有显著性差异,P＜0.05,尿毒灵组、尿毒清组与模型组相比差异显著,P＜0.05,给药后有统计学意义。临床观察部分:尿毒灵疗效:显效5例,占16.67%;有效21例,占70%;无效4例,占13.33%;总有效率86.67%。用药前后对照,患者症候积分下降有统计学意义(P＜0.01)。用药前后对照,患者尿素氮,血肌酐下降有统计学意义(P＜0.05);患者CHOL、TG和LDL-L的水平下降有统计学意义(P＜0.05);安全性指标肝功能用药前后比较无统计学意义,血常规白细胞、红细胞无明显变化,血红蛋白有所上升。4.结论经实验研究得出如下结论:尿毒灵有很好的清除血中肌酐,尿素氮等毒素的作用。,尿毒灵具有很好的降低转化生长因子(TGF-β)、纤溶酶原激活物抑制物(PAI-1)在肾脏小球、间质、小管的表达,抑制TGF-β1、smad3、smad4 mRNA合成的作用。所以可以得出结论尿毒灵可以抑制TGF-β/Smad转导通路。尿毒灵可能是通过抑制TGF-β/Smad转导通路,而达到抑制肾小球、肾间质、肾小管纤维化,从而起到肾保护作用。通过对尿毒灵的临床观察,对观察指标及安全性指标的前后对比。尿毒灵可以很好的改善糖尿病肾病肾功能不全患者临床症状,清除血中小分子毒素,增加患者血红蛋白的作用;未发现有明显的降血尿酸作用;未发现患者有任何不良反应,及不良事件的发生,毒副作用不明。更多还原

【Abstract】 1.Objective and significanceThe purpose of this study is the clinical effect of uremic Ling observed in strict accordance with the diagnostic criteria for inclusion criteria to select patients with drug use,and observe the points syndromes,biochemical indicators such as changes in;At the same time,the role of Hope for the uremic mechanisms to TGF/smad pathway targets for research,experimental study of exploratory;aimed at finding new treatments for DN on the experience of veteran experts who carried out the role of the clinical efficacy and mechanism at the same time,in order to prepare for the development of new drugs.2.MethodInpatient and outpatient diabetes were selected in line with the diagnostic criteria of DN patients,in line with the diagnostic criteria,serum creatinine 178≤(Cr)≤442ummol/l,to exclude other causes of renal dysfunction observed in patients with 30 cases observed for three months observed in patients with symptoms of drug use before and after integration, biochemical,blood lipids,liver function,blood changes,and the final effect.SPF level 48 male Wister rats,weighing 180-220g,adaptive feeding week. Were randomly divided into normal control group 12,36 for the remainder of model.Selected as the model way to STZ(streptozotocin) + adenine ig:STZ used to buffer(2.1g of citric acid dissolved in 100ml double distilled water pair A liquid,2.44g sodium citrate dissolved in 100ml double distilled water for the B solution,A,B buffer solution is mixed) configuration,36 rats were fasted for 12 hours,STZ with a dose of 60mg/kg body weight,intraperitoneal injection of one-time,three days Random post-test blood glucose,blood glucose is greater than 16.1mmol/l of the rat,model successful,To reduce the mortality,rate of blood glucose greater than 25mmol/l in rat subcutaneous insulin ordinary 1-2iu,inadequate blood glucose 16.1mmol/l in rats, complement injection STZ 20mg/kg;all into a mold,the 2 percent to adenine, 300mg/kg administered every 7 days until the check into one orbital venous mode..Model the process five rats dead,after the mold into the remaining 31 rats,high to low according to serum creatinine levels,divided into model group,Niaoduling group、Niaoduqing group;model group,2-4ml orally given normal saline,NiaoduLing group to 0.432g/kg,Niaoduqing Niaoduqing group to 0.27g/kg;administration two months after the orbital venous plexus biochemical blood test,so take some kidney immunohistochemistry,kidneys do some RT-PCR.3.ResultExperimental study components:treatment group compared with the model group,serum creatinine uremic spiritual group,blood urea nitrogen were statistically significant decline;Ling uremic group compared with the model group of cholesterol,triglyceride,low-density lipoprotein cholesterol were no significant differences,no statistical significance;TGF-β1,Smad3, Smad4,PhI-1 expression in model group than those in blank control group significantly increased,with statistical significance,P＜0.05,Ling uremic group than those in the above-mentioned protein expression in model group decreased significantly;RT-PCR detection of rat renal tissue TGF-β1mRNA, Smad3 mRNA,Smad4 mRNA,fluorescence quantitative PCR detection of various targets in the kidney tissue,the model group and blank group were significantly different,P＜0.05,Ling-uremic group Niaoduqing group compared with the model group significantly different,P＜0.05,statistically significant after administration.Clinical Observation Part:Effect of uremic Ling:5 cases markedly,accounting for 16.67%;21 cases effective,accounting for 70%;nulland void four cases,accounting for 13.33%; the total effective rate 86.67%.Control before and after medication,the patient symptom score was statistically significant decrease(P＜0.01).Before and after contrast administration,patients with blood urea nitrogen,serum creatinine was statistically significant decrease(P＜0.05); patients with CHOL,TG and LDL-L reduction in the level of statistical significance(P＜0.05);the safety of drug use indicators of liver function no statistical significance before and after,blood leukocytes,no significant changes in red blood cells,hemoglobin increased.4.ConclusionExperimental studies by the following conclusion:a good soul uremic serum creatinine clearance,blood urea nitrogen,such as the role of toxins.Uremic Ling has a good reduction of transforming growth factor(TGF-β),plasminogen activator inhibitor(PAI-1) in the kidney of small ball,interstitial,tubular expression of TGF-β1 inhibition of mRNA synthesis.Therefore,it can be concluded that uremic TGF-β/Smad Ling can inhibit transduction pathway.Ling uremic TGF-β/Smad may be through inhibiting the transduction pathway,to achieve the inhibition of glomerular,renal interstitial fibrosis,tubular fibrosis,and thus play a role in renal protection.Through clinical research,to observe the targets and safety indicators before and after contrast.A very good effect can be gained,renal failure symptoms in patients with uremic diabetic nephropathy was improved,removal of small and medium-sized molecules of blood toxins,to increase the role of hemoglobin;not found a clear role in lowering serum uric acid;not found any adverse reactions in patients,and adverse events the occurrence of unknown side effects.更多还原