目的:观察中药“复方鳖甲软肝片方”对TGF-β1诱导的肾间质成纤维细胞(NRK-49F)增殖及分泌细胞外基质的影响。方法:以2 ng/ml hTGF -β1诱导 NRK -49F 细胞,并以软肝片方大、中、小剂量(分别以7g/kg、3.5 g/kg、1.75 g/kg)药物血清进行干预,分别应用MTT法、ELISA、免疫细胞化学方法,观察肾间质成纤维细胞增殖、细胞上清FN浓度及ColⅠ、ColⅢ的表达。结果:软肝方含药血清对hTGF-β1诱导的细胞增殖无影响;但大、中剂量软肝方能不同程度地降低FN的分泌及ColⅠ、ColⅢ的表达,软肝方大剂量组作用最强(P〈0.01)。结论:复方鳖甲软肝片方在一定程度上抑制TGF-β1诱导的肾间质成纤维细胞ColⅠ或ColⅢ的表达,大剂量软肝方作用最强。
Objective:Objective:in vitro,we object the effect of Compound Biejia Ruangan Precription( CBRP) on renal in-terstitial fibroblast proliferation and excreting extracellular matrix induced by TGF-β1 . Methods:Using method of serological pharma-cology, rat renal interstitial fibroblast (NRK-49F cell) stimulated with TGF-β1, simultaneously 10% different rat serum was add-ed to for 48 h as follows:the normal rat serum( male SD rats were given physiological saline by gastric gavages) , the low dose CBRP-medicated rat serum (given CBRP 7 g/kg), the middling dose one(given CBRP 3. 5 g/kg), the high dose one(given CBRP 1. 75 g/kg), applying to immunocytochemistry, MTT assay, ELISA, cell proliferation and extracellular matrix such as fibronectin, typeⅠcollagen, typeⅢ collagen were detected. Results:Contrasted with the normal rat serum: rat serum medicated-CBRP had no effect on NRK-49F cell proliferation which induced by TGF-β1 , but in the groups of rat serum medicated- high dose CBRP, the high level of cell secretion fibronectin, collagenⅠor collagenⅢwhich boosted by TGF-β1 had been decreased, especially the group of high dose CBRP had best effect (P〈0. 01). Conclusion:In some extent CBRP could restrain renal interstitial fibroblast secrete or synthesize extracellular matrix such as FN, Col, ColⅢwhich enhanced by TGF-β1 , especially the high dose CBRP exert preferable effects.