【作者】 黄吉峰；

【导师】 李显筑；

【作者基本信息】 黑龙江中医药大学 ， 中西医结合临床， 2010， 博士

【摘要】 目的：本课题通过实验研究,观察肾白宁对糖尿病肾病模型大鼠的影响,探讨糖尿病肾病的中医发病机制及其肾白宁的作用机理,为临床合理用药奠定一定的基础。方法：研究采用左侧肾脏摘除+STZ诱发糖尿病肾病的大鼠模型,中药组将肾白宁浸膏溶于生理盐水,配制成混悬液,以2.97g/kg(含生药量),1次／天,灌胃,2ml/次；将阳性对照组洛汀新溶于生理盐水,配制成混悬液,予以洛汀新0.9mg/kg,1次／天,灌胃,2ml/次。空白对照组及假手术组予以同体积的生理盐水溶液2ml,1次／天,灌胃。观察肾白宁对其尿微量蛋白排出量、尿NAG.ACR.实验鼠的肾重量及体重量的变化情况；采用免疫组化、RT-PCR及彩色病理图像分析系统,定性与定量检测肾组织TNF-α、Bax.Bcl-2.Co-Ⅳ、LN、FN及CTGFmRNA的表达情况；以原位末端标记法检测细胞凋亡情况。结果：12周后光镜下可见空白对照组肾小球毛细血管球萎缩、球囊腔略增大、肾小球出入球小动脉有轻度内皮细胞增生及纤维化、基底膜略增厚,远曲小管扩张,肾间质充血、水肿、血细胞浸润、并有轻度间质纤维化。中药组与阳性对照组比较,体重量的增加程度明显优于阳性对照组,肾脏体重指数均呈下降趋势,但差异不显著。中药组在第12周时与假手术组、空白对照组相比较尿微量白蛋白、NAG、ACR有显著性差异；中药组TNF均有显著降低；肾小球中Bax、Bcl-2表达各组几乎为阴性,肾小管中Bax、Bcl-2表达假手术组显著低于其他三组,中药组与空白对照组比较细胞凋亡率显著降低、Bax阳性表达低,而Bcl-2表达高,中药组与阳性对照组比较无明显差异。空白对照组、中药组、阳性对照组肾组织CTGFmRNA较假手术组高,有显著性差异,中药组、阳性对照组与空白对照组比较阳性表达低,中药组与阳性对照组有显著性差异。结论：肾白宁可增加实验鼠体重量,改善肾重／体重的比值,能明显改善尿微量白蛋白蛋白定量、NAG.ACR,显著降低肾组织CTGFmRNA含量,TNF-α含量,能明显抑制肾组织细胞凋亡、能明显降低Bax表达,提高Bcl-2表达,保护肾小球和肾小管结构,改善肾功能。更多还原

【Abstract】 Objective:We observed systematically curative effect of preventing and treating Diabetic nephropathy(DN) rats by Shenbaining through clinical and experiment researches.We inquired into mechanism of traditional Chinese medicine(TCM) on DN and mechanism of Shenbaining on Preventing and treating DN.Methods:In the study,we use the left kidney of the STZ-induced DN rat.The TCM group will dissolve shenbaining in saline, prepared into suspension and it was given 2.97g/kg,1 time/day, orally,2ml/time. The positive control group lotensin was prepared into a suspension,and also was given 0.9mg/kg,2ml/time, 1time/day, orally.Control group and the sham group to the same volume of saline solution 2m,ltime/day.Then we observed the condition of their urine protein excretion,urine NAG,ACR,rat kidney weight and body weight changes.Using immunohisto chemistry,RT-PCR and color image analysis system to qualitative and quantitative detection of kidney TNF-α,Bax,Bcl-2,ColⅣ,LN, FN and expression of CTGF mRNA.Detect cell apoptosis with the original end-labeling method.Results:12 weeks later,it can be found in the control group that glomerular atrophy ball, balloon chamber slightly increased the glomerular afferent artery with slight endothelial cell hyperplasia and fibrosis,basement membrane thickening slightly,distal convoluted tubule expansion,renal interstitial congestion,edema,blood cell infiltration and mild interstitial fibrosis.Compared with the positive control group,the TCM group’s body weight increase was significantly better,the kidney weight index were decreased,but the difference was not significant. At the 12th week,the TCM group urine albumin,NAG,ACR was significant different from the control group and the sham group;TNF condition of the TCM group were significantly lower with;Glomerular Bax, Bcl-2 expression of almost all groups were negative,tubular in Bax, Bcl-2 expression in sham group was significantly lower than the other three groups,the TCM group and the control group had significantly lower apoptosis,Bax expression is low as the expression of Bcl-2 higher,the TCM group and the positive control group showed no significant difference. The control group,the TCM group and the positive control group′s renal CTGF mRNA were higher than the sham operation group, there were significant differences.The TCM group,positive control group and blank control group expression is low,There was a significant difference between the TCM group and the positive control group.Conclusion:Shenbaining can increase the DN rat’weight, improve the kidney weight/body weight ratio,significantly reduced renal CTGF mRNA content, TNF-αlevels,improve urinary albumin protein, NAG, ACR,inhibit renal apoptosis,decrease the expression of Bax,Bcl-2,increased the expression of glomerular and tubular structure to protect and improve renal function. NAG,ACR and microalbuminuria are more sensitive than the response to renal injury in control of renal hypertrophy.更多还原