| Purpose:In this study, we observe renal AQP1 and tumor necrosis factor-α(TNF-α) expression changes in the rats that are set up as the model of chronic renal failure with kidney-yang deficiency, according to biological and morphological technique, in order to explore the Pharmacological treatment mechanism of zhenwu decotion in disorder of water metabolism of CRF, and to provide experimental evidence for pathophysiological mechanisms of "obstruction of qi activities" of Kidney-Yang Deficiency.Materials and methods:1. Setting up Model and Medicating:Thirty Healthy male wistar rats weighting 200土20g provided by experimental animal center of liaoning university of traditional chinese medicine, adaptive feeding for three days. Then randomized by weight into normal group, model group, zhenwu decotion group with 10 rats in each group. The rats model of chronic renal failure with kidney-yang deficiency was set up by Adenine stomach irrigating. From frist day of experiment, rats of model group and zhenwu decotion group were given by 10ml/kg/d adenine stomach irrigation every morning, while the normal control group rats were given only irrigation stimulability. It lasts 21 days. After two weeks, rats of model group and zhenwu decotion group were given saline solution (at 10ml/kg/d) and zhenwu decotion (atlOml/kg/d) respectively every afternoon besides the Adenine irrigation in the morning for 21 days.2. In experimental process, the general behavior was observed:Body weights were measured to observe changes of average weight. One day before drawing materials, the rats were taken to metabolic cage to reserve 24h urine volumes, in order to measured 24h urine volume, urine specific gravity changes.3. Drawing materials from rats and indexes were detected:Rats were anesthetized by intraperitoneal injection of urethane (at 5ml/kg).2ml blood were taken from abdominal aorta for the detection of Scr, BUN; the renal tissue was plucked off to observe pathological analysis and detection of AQP1, TNF-αby immunohistochemistry method; Plucking off the other side of the kidney, to determine expression of renal AQP1 Western blot analysis.4. Statistical data analysis:The data were expressed as x土SD. Completely randomized design one-way ANOVA was used to deal with data by SPSS13.0 statistical software, P<0.05 for significantly difference.Results:1. Impact of zhenwu decotion on renal function of chronic renal failure rats with kidney-yang deficiencyCompared with normal group, the model group rats appeared intolerance to cold, cold four limbs, loss of appetite, weight loss, less activity, a marked increase in urine volume, decreased specific gravity, increase of blood Scr and BUN content significantly (P<0.05), Renal tissue by light microscopy observation shows glomerular hyperemia, expand cysts, interstitial fibrosis and a large amount of inflammatory cell infiltration, tubular compensatory enlargement. And zhenwu decotion group rats show better feeding, stronger activity, more shiny hair, pretty sencitive reaction, body weight increasing, urine volume decreased, urine specific gravity increased, Scr and BUN content in blood decreased (P<0.05), Renal tissue pathological injury relieved2. Influence of zhenwu decotion on the expression of renal tissue AQP1 of rats of chronic renal failure with kidney-yang deficiencyImmunohistochemical detection shows:Compared with the normal group, AQP1 expression of model group rats in renal proximal tubule epithelial cell decreased and the integrated optical density (IOD) decreased deeply (P<0.05); Compared with model group, AQP1 expression of zhenwu decotion group rats in renal proximal tubule epithelial cell membrane and cytoplasm of kidney connecting duct increased. Imaging analysis showed IOD increased deeply (P<0.05). Western blot proved that AQP1 expression of renal tissue decreased obviously in model group rats. Gray scales was determined with image analysis system. Compared with normal group, gray scales of renal tissue AQP1 expression decreased in model group rats (P<0.05). Compared with model group, AQP1 expression of renal tissue increased obviously in zhenwu decotion group rats with western blot, gray scales of renal tissue AQP1 expression was higher in zhenwu decotion group rats than that in model group (P<0.05)3. Influence by zhenwu decotion to the expression of TNF-a in renal tissue of chronic renal failure rats with kidney-yang deficiency.The normal control group, model group and zhenwu decotion group of interstitial cells of rat kidney membrane and cytoplasm of brown granules can be seen. model group cytoplasm of interstitial cells in the membrane TNF-a expression was significantly increased compared with model group (P<0.05). zhenwu decotion group cytoplasm of interstitial cells in the membrane TNF-a expression was significantly reduced compared with model group (P<0.05).The theory of Chinese medical considered kidney governing water.. It mainly manifested in maintaining the balance of the body water metabolism. During kidney yang deficiency, it will occur obstruction of qi activities and result in disturbance of water metabolism. The experimental results indicate that one of the essence of obstruction of qi activities is AQP1 expression decreased because of increased content and expression of TNF-α, it has influence on renal reabsorb ability.Conclusion:1. Chronic renal failure rats with kidney-yang deficiency have a marked increase in urine volume, decreased specific gravity, increase of blood Scr and BUN content and renal tissue pathological injury significantly. After treatment zhenwu decotion can put it into reverse.2. AQP1 expression of renal tissue of Chronic renal failure rats with kidney-yang deficiency decreased. After treatment with zhenwu decotion, AQP1 expression of renal tissue increased.3. TNF-a expression of renal tissue of Chronic renal failure rats with kidney-yang deficiency increased. After treatment with zhenwu decotion, TNF-αexpression of renal tissue decreased. |
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