Mechanisms of sclerosis in the glomerulus as a representative of microcirculation

在上述基金的资助下，我们对肾小球硬化症的发病机制进行了研究和报道。（1）人肾小球系膜细胞（human mesangial cells，HMC）在重组基底膜上培养时，Matrigel HMC聚集形成孤立的集落，然后延伸出一系列细胞突起，形成树突状网络结构，增殖缓慢。在I型胶原凝胶上，HMC显示出快速的细胞生长。(2)在含有III型胶原、肝素和硫酸乙酰肝素的凝胶上，系膜细胞增殖明显受到抑制; IV型胶原抑制HMC增殖。(3)条件培养液（CM）对系膜细胞的增殖有抑制作用，并可被抗TGF-β抗体吸收。在CM中，少于10%的TGF-β以活性形式存在。直接添加抗TGF-β抗体到培养基中增强系膜细胞增殖。(4)在共培养系统中检测内皮细胞和系膜细胞之间的直接细胞接触对系膜细胞行为的影响。MoAb.1-22-3优先结合于面向内皮细胞的有限系膜细胞表面。(5)MoAb.1-22-3与抗Thy-1.1抗体（OX-7）的反应性存在差异，目前我们正在从细胞（免疫电镜）和分子（利用Thy-1基因cDNA）水平研究MoAb.1-22-3检测的新表位。

With aid of the above grant, we have researched and reported on the mechanisums of glomerulosclerosis. We have obtained the following results during the period of research project :(1) When human mesangial cells(HMC) were cultured on a reconstituted basement membrane, Matrigel HMC aggregated and formed isolated colonies, then extended an array of cell processes to form a dendritic network structure and proliferated very slowly. On type I collagen gel, HMC showed rapid cell growth.(2) Mesangial cell proliferation was inhibited markedly on gels containing type III collagen, heparin and heparan sulphate ; type IV collagen suppressed HMC proliferation.(3) Conditioned medium(CM) of cultured mesangial cells showed the inhibitory activity on mesangial cell proliferation, which was absorbed with anti-TGF-beta antibody. In CM, less than 10% of TGF-beta was present in active form. Direct addition of anti-TGF-beta antibody to culture media enhanced mesangial cell proliferation.(4) The effect of direct cell contact between endothelial and mesangial cells on mesangial cell behavior was examined in a coculture system. MoAb.1-22-3 was bound preferentially to the limited mesangial cell surface facing endothelial cells.(5) There is a difference in reactivity between MoAb.1-22-3 and anti-Thy1.1 Ab(OX-7).Now, we are investigating a new epitope detected with MoAb.1-22-3 at the cellular (immune electronmicroscopic) and molecular (using cDNA for Thy-1 gene) levels.