Experimental Study Of Regulatory Role Of Jian Pi Shen Shi Decoction On Urate Transporter Protein In Gout

Objective：In order to disclose the part molecular mechanisms of the treatmentof gout and hyperuricemia with Jian Pi Shen Shi Decoction, provide theoreticalbasises for it in treatment of gout and enrich theories of preventing and treating goutwith traditional Chinese medicine,we study the regulating effect of Jian Pi Shen ShiDecoction for urate transporter proteins （UAT,URAT1,OAT1and OAT3）in renaltubular epithelial cells of hyperuricemia rat model and the human renal tubularepithelial cells (HK-2cells) cultured in vitro.Methods：1According to serum uric acid levels,48SD rats are divided equallyinto into6groups.That is the control group,the model group, the positivegroup(benzbromarone), Jian Pi Shen Shi Decoction low dose group, Jian Pi Shen ShiDecoction middle dose group and Jian Pi Shen Shi Decoction high dose group. Allgroups except the control group are fed with fodder yeast plus adenine to get thehyperuricemia rat model. After14days, we test uric acid, creatinine and urea nitrogenof the serum of grouped rats,calculate viscera index of the kidney,liver andspleen,observe the pathological injury of rat renal with HE staining of paraffinsections,and measure the expression of urate transporter protein(UAT,URAT1,OAT1and OAT3) in renal tubule of rat model byimmunohistochemistry.2After HK-2cells cultured in vitro were treated by diffierentconcentration of the containing serum of Jian Pi Shen Shi Decoctio or normal salinecontaining serum,we detect the expression of mRNA and protein ofhUAT,hURAT1,hOAT1,hOAT3by RT-PCR and Western blotting. Results：1Three dose groups of Jian Pi Shen Shi Decoction can effectivelyreduce serum uric acid (P<0.05), serum urea nitrogen and creatinine of thehyperuricemia rat model (P<0.05，P<0.01) without liver injury caused by positivedrug benzbromarone. Jian Pi Shen Shi Decoction high dose group could reduce uratecrystal deposition and inflammatory cell infiltration in renal tubule and interstitialsites of hyperuricemia rat model,raise the protein expression of OAT3and couldlower the the protein expression of URAT1, but it has no significant effect of theprotein expression of UAT,OAT1.2For HK-2cells cultured in vitro, compared withthe same concentration of normal saline serum, the5%containing serum of Jian PiShen Shi Decoction can raise the mRNA expression of hUAT and the10%containingserum of Jian Pi Shen Shi Decoction significantly lower the mRNA expression ofhURAT1, while diffierent concentration of the containing serums of Jian Pi Shen ShiDecoctio have no difference with the mRNA expression of hOAT1,hOAT3.5%and10%containing serum of Jian Pi Shen Shi Decoction significantly raise the proteinexpression of hUAT and lower the protein expression of hURAT1, while diffierentconcentration of the containing serums of Jian Pi Shen Shi Decoctio have nodifference with the protein expression of hOAT1,hOAT3.Conclusions:1Jian Pi Shen Shi Decoction can effectively protect the renalfunction and significantly reduce the serum uric acid,which is worth furtherresearch,development and utilization.2Jian Pi Shen Shi Decoction can regulate theprotein expression of URAT1,OAT3in renal tubular epithelial cells of hyperuricemiarat model.3Jian Pi Shen Shi Decoction can regulate the protein and mRNAexpression of hUAT,hURAT1in HK-2cells cultured in vitro.4The upregulation ofexpression of UAT,OAT3and the downturn of expression of URAT1in renal tubulemay be one of the mechanisms in treatment of gout and hyperuricemia with Jian PiShen Shi Decoction.