本文選題：紫癜性腎炎 + 清熱止血顆粒��； 參考：《河南中醫(yī)學(xué)院》2013年碩士論文

【摘要】：【目的】本實(shí)驗(yàn)通過(guò)體外純化HSPN（血尿蛋白尿型）患兒致病性IgA1作用于實(shí)驗(yàn)動(dòng)物腎小球系膜細(xì)胞、足細(xì)胞損傷模型，比較清熱止血顆粒、GTW、清熱止血顆粒+GTW、強(qiáng)的松+肝素四組藥物對(duì)aIgA1刺激后實(shí)驗(yàn)動(dòng)物腎小球系膜細(xì)胞、足細(xì)胞損傷模型TNF-α、nephrin的分泌和表達(dá)的影響；了解清熱止血方、GTW是否通過(guò)阻斷系膜細(xì)胞－足細(xì)胞軸和（或）直接保護(hù)足細(xì)胞而在HSPN治療中起到快速降蛋白尿療效的作用；驗(yàn)證在清熱止血方聯(lián)合GTW應(yīng)用過(guò)程中，清熱止血方、GTW的作用孰輕孰重或是兩藥的協(xié)同作用。 【方法】本實(shí)驗(yàn)通過(guò)制備清熱止血顆粒、GTW、清熱止血顆粒+GTW、強(qiáng)的松+肝素四組實(shí)驗(yàn)動(dòng)物含藥血清；將體外純化的HSPN(血尿蛋白尿型）患兒致病IgA1作用于SD大鼠系膜細(xì)胞、SPF級(jí)小鼠永生化足細(xì)胞，制備HSPN系膜細(xì)胞、足細(xì)胞損傷模型；將各組藥物含藥血清干預(yù)系膜細(xì)胞、足細(xì)胞損傷模型。觀察HSPN患者和健康對(duì)照者血清aIgA1對(duì)系膜細(xì)胞、足細(xì)胞的增殖影響；檢測(cè)系膜細(xì)胞、足細(xì)胞損傷模型及藥物干預(yù)后兩種細(xì)胞損傷模型TNF-a分泌情況；運(yùn)用RT-PCR檢測(cè)足細(xì)胞損傷模型和含藥血清干預(yù)后足細(xì)胞損傷模型的Nephrin mRNA表達(dá)量的變化。 【結(jié)果】我們利用親和層析聯(lián)合分子篩層析的方法,提取了純度較高的mIgA1,并對(duì)其進(jìn)行了鑒定及熱聚合，通過(guò)MTT法發(fā)現(xiàn)由HSPN患兒血清中mIgA1熱聚合后的aIgA1可以引起實(shí)驗(yàn)動(dòng)物系膜細(xì)胞和足細(xì)胞的損傷；通過(guò)ELISA法發(fā)現(xiàn)HSPN患兒aIgA1可以刺激系膜細(xì)胞及足細(xì)胞產(chǎn)生大量的TNF-α；通過(guò)清熱止血方等四組藥物干預(yù)HSPN系膜細(xì)胞、足細(xì)胞損傷模型，發(fā)現(xiàn)清熱止血方+GTW在抑制系膜細(xì)胞增殖中效果明顯，清熱止血方+GTW、GTW、強(qiáng)的松+肝素三組含藥血清抑制系膜細(xì)胞、足細(xì)胞產(chǎn)生TNF-α中較單用清熱止血方效果明顯；四組藥物均可以上調(diào)足細(xì)胞損傷模型（HSPN患者IgA1-系膜細(xì)胞共培養(yǎng)液上清組、HSPN患者IgA1組）nephrin mRNA的表達(dá)，但以清熱止血方+GTW、GTW、清熱止血方三組效果突出，，強(qiáng)的松+肝素在上調(diào)足細(xì)胞損傷模型（HSPN患者IgA1組）nephrin mRNA的表達(dá)中較空白對(duì)照組差異無(wú)統(tǒng)計(jì)學(xué)意義。 【結(jié)論】aIgA是HSPN腎小球系膜細(xì)胞、足細(xì)胞損傷的重要因素之一；腎小球系膜細(xì)胞異常增殖和系膜細(xì)胞、足細(xì)胞產(chǎn)生大量TNF-α是HSPN腎臟損傷的重要表現(xiàn)；清熱止血方、清熱止血方+GTW、GTW、強(qiáng)的松+肝素四組藥物均可通過(guò)阻斷系膜細(xì)胞-足細(xì)胞軸在HSPN（血尿+蛋白尿型）治療中起到快速降低蛋白尿的作用；清熱止血方+GTW、GTW、清熱止血方在直接保護(hù)足細(xì)胞作用方面優(yōu)于強(qiáng)的松+肝素，強(qiáng)的松+肝素在阻斷系膜細(xì)胞-足細(xì)胞損傷軸方面優(yōu)于單用清熱止血方；清熱止血方聯(lián)合GTW應(yīng)用過(guò)程中，在阻斷系膜細(xì)胞-足細(xì)胞損傷軸中GTW的作用優(yōu)于于清熱止血方，在直接保護(hù)足細(xì)胞中二者效果相當(dāng)，總體而言，二者聯(lián)合應(yīng)用優(yōu)于單用一種。
[Abstract]:[objective] to study the effect of purified HSPN (hematuria proteinuria type) IgA1 on glomerular Mesangial cells and podocytes injury model in vitro. To compare the effects of Qingre Zhixue granule (GTW), Qingre Zhixue granule (GTW) and prednisone heparin (prednisone) on the secretion and expression of TNF- 偽 nephrin in glomerular Mesangial cells and podocytes of experimental animals after aIgA1 stimulation. To understand whether Qingre Zhixue prescription GTW plays a rapid role in reducing proteinuria in the treatment of HSPN by blocking Mesangial cell-podocyte axis and / or protecting podocytes directly, and validating the effect of Qingre Zhixue recipe combined with GTW. The action of GTW of clearing away heat and stopping bleeding is important or the synergistic effect of the two drugs. [methods] in this experiment, four groups of experimental animals containing serum of Qingre Zhixue granule (GTW), Qingre Zhixue granule (GTW) and prednisone heparin (prednisone) were prepared. The model of HSPN Mesangial cell and podocyte injury was established by treating the purified HSPN (hematuria proteinuria type) pathogenic IgA1 on rat Mesangial cells of SD rats in vitro, and the serum of each group was used to interfere with the Mesangial cells. Model of podocyte injury. The effects of serum aIgA1 on the proliferation of Mesangial cells and podocytes were observed in patients with HSPN and healthy controls, and the secretion of TNF-a in Mesangial cells, podocytes injury models and two kinds of cell injury models after drug intervention were detected. The changes of Nephrin mRNA expression in podocyte injury model and podocyte injury model were detected by RT-PCR. [results] the mIgA1 with high purity was extracted by affinity chromatography combined with molecular sieve chromatography, and its identification and thermal polymerization were carried out. By MTT method, it was found that the damage of Mesangial cells and podocytes of experimental animals could be caused by aIgA1 after hot polymerization of mIgA1 in serum of children with HSPN. It was found by ELISA method that aIgA1 could stimulate Mesangial cells and podocytes to produce a large amount of TNF- 偽 in children with HSPN. Four groups of drugs, such as Qingre Zhixue prescription, were used to interfere with HSPN Mesangial cells and podocyte injury model. It was found that Qingre Zhixue recipe (GTW) had obvious effect in inhibiting the proliferation of Mesangial cells. The effect of Qingre Zhixue recipe GTW and prednisone heparin on inhibiting Mesangial cells in three groups was more obvious than that of Qingre Zhixue recipe alone. All the four groups could upregulate the expression of nephrin mRNA in the IgA1 group of the patients with HSPN in the IgA1-Mesangial cell co-culture supernatant group, but the effect of the three groups was remarkable. There was no significant difference between prednisone heparin and control group in upregulating the expression of nephrin mRNA in IgA1 patients with podocyte injury. [conclusion] aIgA is one of the important factors of glomerular Mesangial cell and podocyte injury in HSPN. The abnormal proliferation of Mesangial cells and Mesangial cells in glomerular Mesangial cells and the production of a large amount of TNF- 偽 by podocytes are important manifestations of renal injury in HSPN. GTW and GTW, prednisone heparin could reduce proteinuria rapidly by blocking Mesangial cell-podocyte axis in the treatment of HSPN (hematuria proteinuria type). The effects of GTW and GTW on direct protection of podocytes were superior to those of prednisone heparin, and prednisone heparin was superior to Qingre Zhixue recipe in blocking the injury axis of Mesangial cell-podocyte. During the application of Qingre Zhixue recipe combined with GTW, the effect of GTW in blocking the injury axis of Mesangial cell-podocyte was better than that of Qingre Zhixue recipe, and the effect of both was the same in direct protection of podocyte.
【學(xué)位授予單位】：河南中醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R272

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