Mechanism Of Water And Land Two Elixir Treatment Of Proteinuria Of Nephrotic Syndrome In Rats

Objective:Based on the theories of Traditional Chinese Medicine and modem medicine. This experimental research is designed to study the impact of Shuiluerxian Pellet on Adrianmycin-inducded proteined urine in rats and part of the mechanism of action from the perspectives of Hemo-Biochemistry. Histo-pathology and Molecule-Biology.Methods:50 male wistar rats were divided at random into the following groups: the normal group (n=10), the model group(n=10), the high-dosed Shuiluerxian Pellet group (n=10), the low-dosed Shuiluerxian Pellet group (n=10) and the Bennazepril group(n=10). The nephritic syndrome model was made by injecting Adrianmycin once into the tail vein of the rat. The administrations of medicin were as follows: In the normal and model groups, 0.9% saline was poured into the stomach of rat at a dose of 1ml/100g (body weight); In the high-dosed Shuiluerxian Pellet group, high-dosed abstracted Liquid of Shuiluerxian Pellet was poured into the stomach at a dose of 1ml/100g(body weight); In the low-dosed Shuiluerxian Pellet group, low-dosed abstracted Liquid of Shuiluerxian Pellet was poured into the stomach at a dose of 1ml/100g(body weight); In the Bennazepril group, Bennazepril was poured into the stomach at a dose of 0.3mg/100g(body weight). All the administrations metioned above began one week after the models had been made, each medicine was administered once a day, the period of treatment was three weeks.â‘ Observe the general conditions and weight of body; check 24 hour urea protein quantity(24HUPQ), 7days before and after the model had been made, and on the 7th, 14th, 21th day after the administation of related medicine; checkβ2-MG, ALB and IgG in the urine on the 21th day after the administration of medicine.â‘¡24hours after the last administration of medicine, collect blood with the method of excising the eyeball to check Scr, Bun, ALB, TP, TG and TCH. TNF-αand HA were examined by radiommunoassay(RIA). The positive cell numbers of CD₄₄ in peripheral blood were examined through flow cytometer.â‘¢24hours after the last administration of medicine, the rats were killed at once and the left kidneys were removed, from which 50mg of kidney tissues were weighed and the expression of CD₄₄mRNA of the kidney tissues of rats were examined through RT-PCR method.â‘£24hours after the last administration of medicine, the removed kidney tissues, after being dealt with, were made into super-thin slices, which were observed under H-600 penetrating electric microscope; The remaining kidney tissues, which were dyed with HE and PAS after being dealt with, were observed under light microscope.⑤Statistical method: all the data were analysed with SPSS12.0 statistical software, experimental data were indicated with Mean±SD. Quantitative data were analysed with single-factor analysis of Variance, LSD method was adopted for comparison between groups.Results:Seven days after Adrianmycin was injected through tail vein, 24HUPQ of the rats in each model group increased, compared with that of the rats in normal group. Each of the 24HuPQ＞50mg, which showed that the NS models were established successfully.(1)The research of the impact of the abstracted Liquid of Shuiluerxian Pellet on the Hemo-biochemistry and urea protein of the Adrianmycin-induced rats with kidney disease. On the 7th, 14th, 21th day after the administration of medicine, 24HUPQ of the high-dosed, low-dosed and Bennazepril groups was significantly lower than that of the model group(P＜0.01); The urea ALB, ureaβ2-MG and urea IgG quaqntity of the high-dosed, low-dosed and Bennazepril groups were significantly lower than those of the model group. (P＜0.01) 3 weeks after the administration of medicine, the serum ALB and TP of the rats in the high-dosed, low-dosed and Bennazepril groups were examined to be significantly higher that those in the model group (P＜0.01); the serum level of ALB and TP in the high-dosed and low-dosed groups was examined to be higher than that in the Bennazepril group (P＜0.05, low-dosed group P＜0.01). So far as blood lipid is concerned, the results showed that the content of TG and CHOL in the high-dosed, low-dosed and Bennazepril groups was significantly lower than that in the model group (P＜0.01); The level of TG and CHOL in the high-dosed and low-dosed groups was significantly lower than that in the Bennazepril group (P＜0.01). (In the high-dosed group, CHOL P＜0.05). there was no significant difference when the levels of Bun and Scr in different groups were compared with each other (P＞0.05).All these results indicates that Shuiluerxian Pellet could effectively reduce the Proteinuria of the Adrianmycin-induced rats with kidney disease, and could play a positive role in improving the nutrient condition of the Adrianmycin-induced rats and regulating the metabolism of protein and Lipid.(2)The research of the impact of Shuiluerxian Pellet on the TNF-αand HA of Adrianmycin-induced rats with kidney disease; the TNF-αand HA in the high-dosed, low-dosed and Bennazepil groups were significantly lower than those in the model group (P＜0.01); There was no significant difference when the TNF-αand HA in the high-dosed, low-dosed and Bennazepril groups were compared with each other (P＜0.05); The value of TNF-αand HA was in direct proportion to 24HUPQ, urea-ALB, urea-β2- MG and urea -IgG, which shows that Shuiluerxian Pellet could reduce the Proteinuria probably through affecting the regulation web of cytokine and relieving the inflammatory reactions.(3)The research of the impact of Shuiluerxian Pellet on the positive cell numbers of CD₄₄ in peripheral blood of Adrianmycin-induced rats with kidney disease: The positive cell numbers of CD₄₄ in peripheral blood of the rats in the high-dosed, low-dosed and Bennazepril groups were significantly lower than those in the model group (P＜0.01); The positive cell numbers of CD₄₄ in peripheral blood in the high -dosed and low-dosed and low-dosed groups were lower than those in the Bennazepril group (P＜0.05); The positive cell numbers of CD₄₄ in peripheral blood of rats were in direct proportion to 24HUPQ, urea-ALB, urea-β2-MG and urea-IgG (P＜0.01), which shows that Shuiluerxian Pellet could bring down the positive cell numbers of CD₄₄ of the Adrianmycin -induced rats with kidney disease and reduce the inflammatory injury, and therefore could reduce the Proteinuria.(4)The research of the impact of Shuiluerxian Pellet on the expression of CD₄₄mRNA of the kidney tissues of the Adrianmycin-induced tats with kidney disease: The expression of CD₄₄mRNA of the kidney tissues of the rats in the high-dosed, low-dosed and Bennazepril groups was significantly lower than that in the model group (P＜0.01); there was no significant difference when the expressions of CD₄₄ mRNA of the kidney tissues of the rats in different groups were compared with each other (P＜0.05); The expression of CD₄₄ mRNA in the kidney tissues was in direct proportion to 24HUPQ, urea -ALB, urea-β2-MG and urea -IgG (P＜0.01), which shows the probable mechanism at the gene level that shuiluerxian Pellet could reduced the Proteinuria of the Adrianmycin-induced rats with kidney disease and obstruct the inflammatory injuries of the kidney.(5)The pathological research of the effect of Shuiluerxian Pellet on the Adrianmycin-induced rats with kidney disease. Under the light microscope, the slight hyperplasia of mesangium was seen in the model group, in other groups, there was no significant pathological change in glomerulus of kidney and tubule of kidney. It could be seen under penetrating electric microscope that the main patholigical changes of Adrianmycin -induced rats with kidney disease were the widespread fusion of foot protrusion in the glomerulus of kidney and hyperplasia of local mesanguim cells and matrix, and the peripheral oedema around tubular cells of kidney, sometimes the apoptosis of the endothelium cells was found; In the high-dosed,low-dosed and Bennazepril groups the degree of pathological changes in rats was obviously slighter than that in the model group, and only part of the fusion of foot protrusion could be seen.Conclusion:1.This research proved that Shuiluerxian Pellet could effectively bring down the Proteinuria of the Adrianmycin-induced rats with kidney disease, and play a wholesome role in improving the nutrient conditions of Adrianmycin-induced rats and regulating the metabolism of protein and lipid.2.After the treatment with Shuilurexian Pellet the pathological change were significantly relieved, this provided a histo -pathological evidence for the suggestion that Shuiluerxian Pellet could be used to treat Adrianmycin-induced rats with kidney disease and reduce the Proteinuria.3.The mechanism of how Shuiluerxian Pellet could bring down the Proteinuria of the Adrianmycin-induced rats with kidney disease:Shuiluerxian Pellet could reduce the TNF-α,HA and CD₄₄ possitive cell numbers in the peripheral blood of Adrianmycin-induced rats with kidney disease,and through functioning on the web of functional cytokine-cell adhesion molecule- matrix among cells, it could influence the inflammatory and immuning functions of the organisms and then could relieve the occurrence of inflammatory injuries, and therefore could reduce the Proteinuria: In the meantime,through reducing inflammatory reaction,obstructing the activation of cells,interfering with the expression of CD₄₄ and the reaction between CD₄₄ and its ligand,it could reduce the expression of CD₄₄ mRNA of the kidney tissues of the Adrianmycin-induced rats with kidney disease. and therefore it could delay the injury in kidney.