活血化瘀方有效单体对活化的肾成纤维细胞株和系膜细胞株增殖的影响  

Influences of effective monomers in Huoxue Huayu Fang on proliferation of activated renal fibroblast cell line and mesangial cell line

作　　者：王东[1,2] 张江[3] 何立群[2] 

机构地区：[1]安徽中医药大学第一附属医院,安徽230031 [2]上海中医药大学附属曙光医院 [3]合肥师范学院

出　　处：《北京中医药大学学报》2013年第11期762-767,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家科技重大专项(No.2009ZX09311-003);教育部高等学校博士点专项(No.20093107110006)

摘　　要：目的观察活血化瘀方有效单体对活化的肾成纤维细胞株(NRK-49F)和系膜细胞株(HBZY-1)的影响。方法采用体外培养NRK-49F和HBZY-1的方法,分为空白对照组、模型组(TGF-β15 mg/L)、高剂量中药组(TGF-β15 mg/L加中药10 mg/L)和低剂量中药组(TGF-β15 mg/L加中药1 mg/L)。先用2.5 g/L胰蛋白酶-0.2 g/L乙二胺四乙酸消化NRK-49F和HBZY-1细胞株,制成单个细胞悬液,接种于96孔板中。在细胞生长同步化后,加入TGF-β1生长因子,作用24 h后吸去培养板中的培养液,再加入中药单体,培养24 h后,采用MTT法观察24 h中药单体对2种细胞株的作用以及ELISA法对TGF-β的分泌情况。结果与空白对照组相比,加入TGF-β1刺激因子可以促进2种细胞株的活化(P<0.05),以及TGF-β的分泌(P<0.01),加入中药单体后可以抑制活化的细胞株增殖和TGF-β的分泌(P<0.05或P<0.01)。其中,对活化的2种细胞株抑制效果最好的是丹酚酸B,其次是丹酚酸A、大黄酚和大黄酸。结论活血化瘀方中药单体可以抑制活化的肾成纤维细胞株和系膜细胞株的增殖,有效防治肾纤维化,其可能的作用机制与其抑制TGF-β的分泌有关。Objective To observe the influences of effective monomers in Huoxue Huayu Fang on the proliferation of activated renal fibroblast cell line （NRK-49F） and mesangial cell line （ HBZY-1 ）. Methods NRK-49F and HBZY-1 were cultured in vitro and then divided into blank group, model group （TGF-ββ, 5 mg/L）, high-dose group （TGF-β1, 5 mg/L ＋Chinese medicinal, 10 rag/L） and low-dose group （TGF-β1,5 mg/L ＋ Chinese medicinal, 1 mg/L）. NRK-49F and HBZY-1 were digested with 205 g/Ltrypsin-0.2 g/L EDTA for making single cell suspension, and the suspension was inoculated to 96- well plates. After the synchronization of cell growth, TGF-131 was added for 24 hours, then the cultural medium was removed, and finally the monomers of Chinese medicinal was added. After culturing for 24 hours, the effects of Chinese medicinal on NRK-49F and HBZY-1 were observed by using methyl thia-zolyl tetrazolium test immunosorbent assay （MTr）, and the secretion of TGF-[3 was observed by using enzyme-linked （ ELISA ）. Results Compared with blank group, TGF-β1-stimulating factor improved the activation of NRK-49F and HBZY-1 （ P 〈 0. 05 ） and secretion of TGF-β（ P 〈 0. 01 ）. After adding the monomers of Chinese medicinal, the proliferation of activated NRK-49F and HBZY-1 and secretion of TGF-β were inhibited （P 〈 0.05 or P 〈 O. 01 ）. Salvianolic acid B had the best inhibitory effect, and followed by salvianolic acid A, chrysophanol and rhein. Conclusion The monomers of Huoxue Huayu Fang can inhibit the proliferation of activated NRK-49F and HBZY-1 and prevent renal fibrosis, and the possible mechanism is related to the inhibition of TGF-β secretion.

关 键 词：成纤维细胞株 系膜细胞株 体外培养 活血化瘀方 中药单体 转化生长因子-Β 

分 类 号：R285.5[医药卫生—中药学]