| Research objectives:On the basis of inheriting the academic experience of Professor Zhao Shaoqin and Professor Peng Jianzhong and combining the researchprogress of modern medicine on the theory of "Gut-Kidney Axis".By observing the effects of Tongfu Huazhuo Liangxue Recipe on intestine,kidney and uremic toxin in rats with chronic renal failure,to explore the effects of Tongfu Huazhuo Liangxue Recipe on the producti on,absorption and nephrotoxicity of indolephenol sulfate,to elucidate the protective mechanism of this prescription on renal injury induced by indole sulfate.Research methods:1.Literature researchAt the same time,from the ancient literature,we can find out the causes of the formation of dampness,heat and toxicity in the body of the patients with chronic renal failure,by summing up the experience of treating chronic renal failure from the point of view of "dampness,heat and toxicity",we can verify the theoretical significance and superiority of Tongfu Huazhuo Liangxue recipe.2.Experimental research2.1 To explore the protective effect of Tongfu Huazhuo Liangxue Recipe onthe kidney of rats with chronic renal failure.Sixty SD rats were randomly divided into six groups: Sham operation group,model group,Tongfu Huazhuo Liangxue Formula high dose group,Tongfu Huazhuo Liangxue Formula medium dose group,Tongfu Huazhuo Liangxue Formula low dose groupand Niaoduqing group,with 10 rats in each group.Except blank control group,the other groups established 5/6 chronic renal failure rat models by nephrectomy.The model group was given normal saline by intragastric administration according to 12.06g/kg body weight.The low,medium and high dose groups of Tongfu Huazhuo Liangxue Prescription were given 6.04 g/kg,12.06g/kg and 24.12g/kg body weight respectively.The Niao duqing group was given 12.06g/kg body weight Niaoduqing granule solution by intrag astric administration daily,starting from the fifth week,and continuing intragastric administration until the sixteenth weekend.The rats were killed at the 16 th week,Urea nitrogen(BUN),creatinine(SCr),uric acid(UA)and 24-hour urinary protein were measured in each group,and intestinal and renal tissues were removed.The pathological changes of kidney tissue were observed by HE staining.2.2 To explore the mechanism of Tongfu Huazhuo Liangxue Prescription in alleviating renal function injury caused by uremic toxin indole sulfate through different links of "Gut-Kidney Axis".Sixty SD rats were randomly divided into six groups: Sham operation group,model group,Tongfu Huazhuo Liangxue Formula high dose group,Tongfu Huazhuo Liangxue Formula medium dose group,Tongfu Huazhuo Liangxue Formula low dose groupand Niaoduqing group,with 10 rats in each group.Except blank control group,the other groups established 5/6 chronic renal failure rat modelsby nephrectomy.The model group was given normal saline by intragastric administration according to 12.06g/kg body weight.The low,medium and high dose groups of Tongfu Huazhuo Liangxue Prescription were given 6.04 g/kg,12.06g/kg and 24.12g/kg body weight respectively.The Niaod uqing group was given 12.06g/kg body weight Niaoduqing granule solution by intragas tricadministration daily,starting from the fifth week,and continuing intragastric admin istration until the sixteenth weekend.At the 16 th week,rats were killed,the intestinal and renal tissues of rats were removed and the pathological changes of colon tissues were observed by HE staining.The content of Indolesulfate in serum was determined by high performance liquid chromatography with fluorescence detection.The levels of heme oxidase HO-1 in kidney tissues of rats in each group were detected by Elisa method.Microbial community composition in colonic contents was analyzed based on 16 S rRNA gene sequencing.Research Result:1.Indicators of renal functionTwo months later,compared with the model group,the contents of creatinine,urea nitrogen,uric acid and 24-hour urinary protein in the low-dose group of Tongfu Hu azhuo Liangxue Prescription were significantly lower(P <0.01),and the contents of serum urea nitrogen,creatinine,uric acid and 24-hour urinary protein in the high-dose group,the middle-dose group and the low-dose group of Tongfu Huazhuo Liangxue Prescription were significantlylower(P < 0.05).Tongfu Huazhuo Liangxue prescription group has more therapeutic advantages than Niaoduqing group.Three months after administration,the levels of BUN and SCr in serum of model group were higher than those in two months.Compared with model group,the middle and high doses of Tongfuhua Zhuoliangxue Decoction could reduce the level of BUN in serum of model rats(P < 0.01),and the low,middl and high doses could signif icantly reduce the level of SCr(P < 0.01),(P <0.001),(P < 0.001),with significant difference.Quantitative results of 24-hour urinary protein test showed that after three months of administration of Tongfu Huazhuoliangxue Formula,compared with thes ham operation group,rats in the model group showed significant proteinuria(P < 0.001).Compared with the model group,the 24-hour urinary protein of rats in the threedoses of Tongfu Huazhuo Liangxue Formula group decreased in varying degrees(P < 0.001),and the difference was statistically significant.2.Pathological changes of kidney and colonGlomerular vascular loops and perivascular fibrosis in model group.Tongfu Huazhuo Liangxue Decoction group was lighter than model group and Niaoduqing group.Compared with the model group,the number of tubular types in Tongfu Huazhuo Liangxue Formula group and Niaoduqing group decreased.Compared with Niaoduqing group,the number of tubular types and glomerular inflammation in Tongfu Huazhuo Liangxue Formula group decreased.Inflammatory cell infiltration was the main pathological change of intestine.The number and area of inflammatory cells were the largest in model group,followed by Niaoduqing group.The high,middle and low dose groups of Tongfu Huazhuo Liangxue recipe had lower number of inflammatory cells than the model group.Niuduqing group had an effect on the inflammatory state of intestinal cells,but had a worse effect than Tongfu Huazhuo Liangxue recipe.3.The Serum Content of Uremic Toxin Indole SulfateThe level of Indole Sulfate in serum of rats with chronic renal failureat three months was significantly higher than that of the sham operation group(P < 0.001).The low,middle and high Tongfu Huazhuo Liangxue recipe could reduce the content of In dole Sulfate in serum of rats(P < 0.001).4.Intestinal microorganisms in each groupThe number of intestinal species of rats in each group was 18/34/12 in blank group,model group and Tongfu Huazhuo Liangxue recipe group,respectively,these results suggest that this prescription may cause changes of intestinal flora in rats with renal failure.Principal coordinate analysis was used to study the similarity or difference of sample community composition,the results showed that Tongfu Huazhuo Liangxue Prescription group migratedabnormal intestinal flora to normal intestinal flora,and intestinal flora was between model group and blank group.The relative abundance of Bacteroides in intestinal flora of rats in model group was significantly higher than that in sham operation group(P < 0.05).Elisa method was used to detect the changes of HO-1 level in the kidneys of rats in each group.Compared with the sham operation group,the HO-1 level in the kid neys of rats in the model group was significantly lower(P < 0.05).Compared with the model group,the middle dose Tongfu Huazhuo Liangxue recipe could significantly increase the HO-1 level in the kidneys(P < 0.01).Conclusion:1.Tongfu Huazhuo Liangxue Prescription can remove the "dampness,heat and toxicity" accumulated in kidney and triple Jiao in patients with chronic renal failure.2.Tongfu Huazhuo Liangxue Prescription is effective in treating chronic renal failure model rats.Compared with uremic clearance,Tongfu Xiezhuo Liangxue Prescription can alleviate renal injury,improve the accumulation of uremic toxins,especially the intestin al protein-binding toxin indole sulfate,regulate the intestinal flora of rats with chronic kidney disease,thus delaying the progression of chronic renal failure.3.One of the possible mechanisms of Tongfu Huazhuo Liangxue Prescription in treating chronic renal failure is to intervene in the mechanism of renal damage related to indole sulfate in the Gut-Kidney Axis.Tongfu detention reduces intestinal decay,improves intestinal flora imbalance,and then reduces the production of indolephenol sulfate in the intestine;removes turbidity and detoxification,restores the function of intestinal mucosal barrier,reduces the toxin from the intestine into the blood,there by reducing the content of blood toxicants;cools blood and detoxifies,and inhibits the renal toxicity of indolephenol sulfate. |
PDF Full Text Request