Heat-clearing And Hemostatic, Ra Masato Glycosides By In Vitro Study Of Glomerular Mesangial Cells, Foot Cell Axis And TNF-α, Nephrin Treatment Of Urinary HSPN Protein Intervention

Objective The purification of Henoch-Schonlein purpura nephritis(hematuria and proteinuria type) the effect of IgA1on experimental animalglomerular mesangial cells, to observe the effect of Qingre Zhixue Granules, RaMasato was, Qingre Zhixue Granules+Ra Masato wilfordii, prednisone plusheparin four groups of drugs on experimental animal after stimulation ofmesangial cells, podocytes damage model of TNF-α, nephrin secretion and theexpression;Understanding of Qingre Zhixue party, whether GTW by blockingthe mesangial cells and podocytes axis and (or) directly protect podocyte inHSPN treatment to reduce proteinuria curative effect;Confirmed in QingreZhixue decoction combined with GTW in the application process, heat-clearingand hemostatic prescription,the role of GTW is important or synergistic effectsof two drugs.Methods The purification of Henoch-Schonlein purpura nephritis(hematuria and proteinuria in children with type IgA1) pathogenic role inmesangial cells of SD rats, SPF mouse immortal podocyte preparation HSPNmesangial cells, podocytes injury model.Taking SD rats as the experimentalanimal preparation Qingre Zhixue Granules, Ra Masato was,Qing re Zhi xueGranules+Ra Masato wilfordii,prednisone plus heparin four groups containingserum on cell injury model.Observation of mesangial cells, podocytes morphology, secretion was examined by TNF-a ELISA method, expression ofnephrin protein was detected by indirect immunofluorescence distribution, wasused for detecting Nephrin nucleic acid Quantitative real-time PCR, Westernblot to detect nephrin protein expression.Results Using affinity chromatography and molecular sievechromatography, extraction of high purity mIgA1, and has carried on theappraisal and thermal polymerization; the inhibition of aIgA1can induce cellproliferation and mesangial cells were found by MTT method.By the method ofELISA found that aIgA1can stimulate mesangial cells and podocytes secretionof TNF-α in large Quantities, and caused the mesangial cell podocyte injurymodel. Through the heat hemostatic prescription Qingre Zhixue Fang,+TW, TW,prednisone plus heparin four groups of drug intervention of mesangial cells,podocytes injury model, proved that heat hemostatic+TW inhibit mesangial cellproliferation effect.Four groups of drugs could inhibit the secretion of TNF-αcell model, but with heat-clearing and hemostatic party+TW, TW two groupseffect is outstanding; four groups of drugs were upregulated in podocytes(HSPN IgA1-in patients with mesangial cell co-culture supernatant group) theexpression of nephrin mRNA, but with Qing re Zhi xue party+TW,heat-clearing and hemostatic two group the effect is more outstanding.Fourgroups of drugs were upregulated in podocytes (HSPN patients in IgA1group)the expression of nephrin mRNA, but with+TW, TW, heat-clearing andhemostatic prescription Qingre Zhixue recipe three groups effect.In addition,the western medicine group and prednisone plus heparin in the inhibition ofmesangial cells, podocytes damage model of secretion of TNF-α increased,podocyte injury model (supernatant group and HSPN group IgA1patients werecultured with HSPN in patients with IgA1-mesangial cells) are effectiveexpression of nephrin in mRNA, special effects, but no obvious difference. Conclusion One of the main factors of aIgA are the podocyte injurymesangial cell line, HSPN； TNF-α secretion is an important manifestation ofglomerular mesangial cells, podocytes injury; Qingre Zhixue recipe, heat-clearingand hemostatic prescription+GTW, GTW, prednisone plus heparin four groups byblocking the mesangial cells and podocytes axis in HSPN (hematuria and proteinuriatype) treatment to reduce proteinuria effect of Qingre Zhixue;+GTW, GTW,heat-clearing and hemostatic in direct protection of podocyte function is better thanprednisone plus heparin, prednisone plus heparin in preventing mesangial cells andpodocytes is better than single use Qingre Zhixue recipe; Qingre Zhixue decoctioncombined with GTW in the application process, in preventing mesangial cell andSertoli cell axis in GTW effects due to heat hemostatic, rather, the two effects in thedirect protection of podocytes in general, the two coordinated application is superiorthan single use of one.