The Effect Of Qingrezhixue Recipe And GTW On Mesangial Cell-podocyte Axis Injury Caused By IgA1 In Children With HSPN

Henoch-Schonlein purpura nephritis(HSPN)is one of the main manifestations of Henoch-Schonlein.It is the most important factor of prognosis of Henoch-Schonlein.In our country,HSPN is at the head of the secondary kidney disease in children.In recent years,its incidence has a rising trend.It serious harm to children’ s health.The main pathological changes of HSPN is IgA depositing in mesangial area and mesangial cell proliferation,accompanied podocyte swelling and falling off,or fusion of podocyte foot processes.These lesions is closely related to the level of albuminuria,and it is one of the important factors of disease and prognosis.So intervention of proteinuria is the key to the clinical outcome of HSPN.Till now there are some article reports about the damage of mesangial cells by IgAI,but the damage of podocyte and mesangial cells and podocyte axis is not reported.The subject of professor Ding reported that Qingre Zhixue granule combined GTW can reduce the level of albuminuria.In order to further study we conduct the research.This study,we used the algAI of HSPN in children’S patient to stimulated mesangial cells and podocyte and discussed the damage mechanism.we established the damage model of Mesangial cells and podocyte axis cultured with conditioned medium prepared from MES incubated with aIgA1.We Comparised the influence of pharmacological intervention by Qingre Zhixue granule combined GTW,Qingre Zhixue granule,GTW and prednisone combined heparin with the damage modle of mesangial cells and podocyte axis.To observe the affect of cell proliferation,tumor necrosis factor-α(TNF-α)production and the expression of mRNA of podocyte molecules,we discuss the possible mechanism to reduce the proteinuria by using the four groups of medicine to intervene the damage modle of mesangial cells and podocyte axis.From this research we want to provide some support for the clinical application and experimental basis.Objective:To study the damage mechanism of using algAl of HSPN in children’s patient to stimulated mesangial cells and podocyte.To discuss the affect of cell proliferation,tumor necrosis factor-α(TNF-α)production and the expression of mRNA of podocyte molecules by interving the damage modle of mesangial cells and podocyte axis for Qingre Zhixue granule and GTW.Methods:We were purified the mIgA1 and pIgA1 from the serum of HSPN children’ s patients and healthy children by Jacalin affinity chromatography,and compared the difference of two groups for concentration.Then we adopted the method of thermal polymerization and ultrafiltration concentration aggregated mlgA1 into aIgA1.With different concentrations(0,50,100,250,500,750,500 ug/ml)of aIgA1 from HSPN and healthy children,we stimulated mouse mesangial cells(MES)and mouse podocyte cells(MPC),and contrasted to observe the effects of different concentration aIgA1 on cells proliferation and the changes of secretion level for TNF-α.Using the conditioned medium prepared from MES incubated with aIgA1(100 ug/ml)for 48 hours(aIgA1 MES conditioned medium)stimulated podocyte,contrast with aIgA1 form HSPN and healthy children directly stimulated podocyte.Then observed the effects of cell proliferation,the changes of secretion level for TNF-α,and the expression of mRNA of podocyte molecules such as nephrin,podocin and podocalyxin.Preparing 4 groups of medicated serum,include Qingre Zhixue granule combined GTW,Qingre Zhixue granule,GTW and prednisone combined heparin.Using the medicated serum to intervene the damage modle of mesangial cells and podocyte axis,then observed the effects of cell proliferation,the changes of secretion level for TNF-α,and the expression of mRNA of podocyte molecules.Results:1.The concentration of mIgA1 and pIgA1 with HSPN children’s patient in serum was higher than healthy children.2.The aIgA1 of Children’ s patients with HSPN can stimulate the proliferation of mesangial cells,and stimulated it secrete more TNF-α,and the results is more apparent with the concentration increased in the beginning,and when it reached a certain concentration the proliferation was restrained;The aIgA1 of Healthy children in high concentration can stimulated the proliferation of mesangial cells.3.Two groups of aIgA1 in low concentration can not effected podocyte directly.It can’t effected not only the proliferation but also the level of TNF-α;But both them can effected the cells in high concentration.4.We succeed established the damage model of mesangial cells and podocyte axis by using the conditioned medium prepared from MES incubated with aIgA1(aIgA1 MES conditioned medium)stimulated podocyte.The aIgA1-MES conditioned medium inhibited the proliferation of podocyte,and secreted more TNF-α,and reduced the expression of mRNA with nephrin,podocin and podocalyxin.5.Qingre Zhixue granule combined GTW,GTW and prednisone combined heparin improved the proliferation inhibition of cells,but Qingre Zhixue granule could not;Four groups of drug serum significantly inhibited the increasing of TNF-α and have equivalent efficacy indepression;Qingre Zhixue granule combined GTW,GTW and prednisone combined heparin improved the expression of mRNA with nephrin,podocin and podocalyxin but Qingre Zhixue granule could not.Conclusion:The concentration of both mIgA1 and pIgA1 with HSPN children’ s patient in serum were increasing.The aIgA1 could stimulate the mesangial cells to increased proliferation of cells and secretion of TNF-α.But it shouldn’ t effected podocyte directly.It stimulated podocyte through the mesangial cells and podocyte axis to make the level of TNF-α rasing and related the expression mRNA of molecular.Traditional Chinese medicine Qingre Zhixue granule combined GTW could improve the damage model of mesangial cells and podocyte axis and reduced the secretion of TNF-α and raised expression mRNA of molecular,Inoder to reduce the level of albuminuria.as to reduce the effectiveness of proteinuria.In this link,GTW played a main role.