原发性肾病综合征患儿外周血单个核细胞对肾小球上皮细胞和系膜细胞的直接影响

摘要

Objective　To investigate the effect of immune cell from idiopathic nephrotic children on extracellular matrix (ECM) synthesis by cultured rat glomerular epithelial cell (GEC) and on the proliferation of mesangial cell (GMC). rnMethods　Twenty-eight children with idiopathic nephrotic syndrome and 15 age-matched healthy children were randomly selected and divided into 4 groups: Group 1, untreated nephrotic children; Group 2, glucocorticoid treated nephrotic children; Group 3, children undergoing glucocorticoid treatment with negative proteinuria; and Group 4, normal control. The peripheral blood mononuclear cells (PBMC) were collected from these children and PBMC conditioned medium (PBMC-CM) were prepared. The PBMC-CM was co-cultured with GEC and GMC respectively. The concentrations of collagen, laminin, collagen Ⅲ and collagen Ⅳ in the GEC and PBMC-CM co-culture medium were investigated. The GMC proliferation was measured by the 3　H-thymidin incorporation method. rnResults　The 3 H-proline incorporation coefficients of the GEC treated with the PBMC-CM of the 4 groups were 0.93, 1.24, 1.23, and 1.11, respectively. The laminin inhibitory coefficients of the 4 groups were 0.95, 1.02, 1.01, and 1.04, respectively. The inhibitory coefficients of collagen Ⅲ were 0.97, 1.00, 0.99, and 1.01, respectively, for the 4 groups. All these parameters showed a significant difference between Group 1 and the other 3 groups (P<0.05). However, there was no significant difference in the inhibitory coefficient of collagen Ⅳ between each two of the 4 groups (1.04, 1.05, 1.04, 1.08, P>0.05). The 3　H-thymidine incorporation coefficients of GMC responsive to PBMC-CM were 1.21, 1.53, 1.50, and 1.10, respectively, and no significant difference was found between the 4 groups (P>0.05). rnConclusion　The results suggested that the circulating immune cells from idiopathic nephrotic children have a direct effect on some ECM component synthesis in cultured rat GEC; the bio-activity of immune cells could be neutralized by administering glucocorticoid; and the circulating immune cells of nephrotic children have no direct effect on GMC proliferation.%目的明确原发性肾病患儿的免疫细胞对培养的大鼠肾小球固有细胞的直接作用。rn方法肾病患儿28名，根据皮质激素治疗与否、尿蛋白转阴与否分为未治组、治疗组、尿蛋白阴转组，正常对照组为健康体检儿15名。制备外周血单个核细胞条件液（PBMC-CM），运用肾小球细胞体外培养技术、同位素掺入技术、放射免疫竞争性抑制法，检测1、PBMC-CM刺激后的大鼠肾小球上皮细胞合成层粘连蛋白（LN）、总胶原、Ⅲ型前胶原、Ⅳ型胶原等基质成份的含量；2、PBMC-CM 对大鼠的肾小球系膜细胞3H-TdR掺入指数的影响。rn结果 1、四个观察组的LN抑制指数为0.95、1.02、1.01、1.03；3H-羟脯氨酸的掺入指数为0.93、1.24、1.23、1.11；Ⅲ型前胶原的抑制指数为0.96、1.00、0.99、1.01；Ⅳ型胶原抑制指数为1.04、1.05、1.04、1.08；显示：未治组患儿PBMC-CM作用后的上皮细胞合成LN、总胶原、Ⅲ型前胶原的指数与其它三组有显著差异（P<0.05)；2、四组观测儿的PBMC-CM作用后的大鼠系膜细胞的3H-TdR掺入指数分别为1.21、1.53、1.50、1.10，四组指数之间无显著差异（P>0.05）。rn结论原发性肾病患儿的免疫细胞产物具有影响上皮细胞合成部分细胞外基质的活性，此活性可被整体使用皮质激素纠正；免疫细胞对系膜细胞的增殖功能无直接影响。