| Objective:To explore the mechanism of uric acid nephropathy and the treatment of clear heat and drain dampness for this disease; to explore whether Bixiechubi decoction could interfere the uric acid nephropathy by down-regulating the PI3K/AKT/mTOR pathway to reduce the expression of NLRP3 inflammasome.Methods:The theoretical research was retrieving and analyzing relative documentation to explore the etiology and pathogenesis of uric acid nephropathy. The experimental research was divided into three parts. The first part was to screen the optimal rat for model of uric acid nephropathy. Differences of three kinds of rat model of uric acid nephropathy were compared by several aspects as renal mass, xanthine oxidase, serum uric acid, creatinine, urea nitrogen, and renal pathology, ect. The level of kidney damage was observed under different modeling methods for rats of uric acid nephropathy to select the best animal model for the subsequent experiment. Secondly, six groups of modeling experimental animals were established as blank group, model group, allopurinol group, and three Bixiechubi decoction groups from high, median, and low doses. Model animals were intragastric administrated with adenine and amiodarone. The general condition of the rats and renal pathology were observed with score. The serum uric acid, creatinine, and urea nitrogen of rats were examined, and the protein of NLRP3, caspase-1, ASC, IL-1β were measured by Western blot and their mRNA expression were detected by RT-PCR. Thirdly, the rat macrophage cell NR8383 and its descendants were routinely cultured. Serum contained different concentrations of Bixiechubi decoction was interfered after LPS stimulation was applied onto NR8383. The existence of IL-1β in cell supernatant was examined by EL1SA, the protein expression of p-PI3K, p-AKT, p-mTOR, NLRP3, caspase-1, and ASC wereexamined by Western blot, and the mRNA expression of NLRP3, caspase-1, ASC, IL-1β were examined by RT-PCR.Results:1 Results of theoretical researchThe basic pathogenesis of uric acid nephropathy is Qi stagnation by dampness-heat, causing meridian blockage, belonging to the "pattern of kidney excess". According to meta-analysis, treating this disease by applying clear heat and drain dampness can effectively reduce the indexes of serum uric acid, creatinine, urea nitrogen, and urinary protein, etc. of the patients to protect the renal function. Modern medical research provides that this disease is closely relative to the pathway of PI3K/AKT/mTOR and its downstream NLRP3 inflammasome. Treating uric acid nephropathy by interfered this pathway could be an important mechanism.2 Results of experimental research2.1 Applying adenine and ethambutol can increase the level of xanthine oxidase and uric acid of rat, and enhanced its expression of creatinine and urea nitrogen. Pathological results revealed that lots of uric acid crystal deposition appeared in renal tissue with obvious infiltration by macrophage.Bixiechubi decoction can effectively reduce the serum xanthine oxidase, uric acid, creatinine, urea nitrogen, and the active oxygen in renal tissue to improve the renal pathology of rats and reduce its pathological score, and the protein and gene expression of NLRP3, caspase-1, ASC, and IL-1β.Bixiechubi decoction can reduce the phosphorylation protein expression of PI3K, AKT, and mTOR in NR8383 cell, and decrease its protein and gene expression of NLRP3, caspase-1, ASC, and IL-1β.Conclusion:For theoretical aspect, the mothod of clear heat and drain dampness is an important way to treat uric acid nephropathy for reducing serum uric acid effectively and protecting the renal function. For the experimental aspect, adenine and ethambutol can be applied to create decent model of uric acid nephropathy. Bixiechubi decoction could reduce the xanthine oxidase, uric acid, creatinine, and urea nitrogen of the rat model to protect the renal function and the expression of inflammatory target NLRP3 inflammasome. Cell experiments confirmed that Bixiechubi decoction can alleviate the inflammatory reaction in kidney by reduce the phosphorylation of PI3K, AKT, and mTOR, and reduced the expression of NLRP3. |
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