摘要
目的:观察丹芍化纤胶囊(DSHX)含药血清对大鼠肝星状细胞(HSC)Gremlin和骨形态发生蛋白7(BMP7)表达的影响,探讨该药治疗肝纤维化的可能机制。方法:将20只雄性Wistar大鼠随机分为4组,每组5只,分别经DSHX低剂量(0.5 g·kg-1),DSHX高剂量(1.0 g·kg-1),扶正化瘀胶囊(0.8 g·kg-1)及生理盐水ig 5 d,2次/d,其后股动脉取血制备大鼠DSHX含药血清、扶正化瘀胶囊含药血清及生理盐水血清,4组血清分别干预体外培养的HSC-T6,用MTT比色法测定各组HSC的增殖效应,ELISA法测培养液上清Ⅰ型胶原(ColⅠ),Ⅲ型胶原(ColⅢ)及α-平滑肌肌动蛋白(α-SMA)的含量,免疫细胞染色法检测各组HSC内转化生长因子-β1(TGF-β1)的表达,采用RT-PCR法检测各组HSC Gremlin,BMP7 mRNA的表达,Western blot检测各组HSC Gremlin,BMP7蛋白的表达。结果:与生理盐水血清组比较,DSHX低、高剂量组含药血清均可显著抑制大鼠HSC的增殖(P<0.05),抑制率与扶正化瘀胶囊血清组相当;DSHX低、高剂量血清组培养液上清ColⅠ,ColⅢ与α-SMA的含量明显减少(P<0.01,P<0.05);HSC内TGF-β1蛋白的表达强度显著降低(P<0.01),与扶正化瘀胶囊血清组相当。与生理盐水血清组比较,DSHX低、高剂量血清组Gremlin mRNA及蛋白表达显著下降,而BMP7 mRNA及蛋白表达显著增多(P<0.01),与扶正化瘀胶囊血清组相当。结论:丹芍化纤胶囊可抑制体外培养的大鼠HSC增殖与活化,减少细胞外基质的合成,对大鼠肝纤维化有潜在的治疗作用,其机制可能与下调HSC Gremlin,上调BMP7的表达有关。
Objective：To investigate the effect of Danshao Huaxian capsule （DSHX） on the expressions of Gremlin and bone morphogenetic protein 7 （BMP7） in rat hepatic stellate cell （HSC） and to explore its possible mechanism in treating hepatic fibrosis.Method：A total of 20 male Wistar rats were randomly divided into control group,Fuzheng Huayu capsule group （0.8 g · kg^-1）,low-and high-dose DSHX treated group （0.5,1.0 g·kg^-1） （n =5）.Rats in each group were intragastrically administered with the corresponding medicines,respectively,twice per day for 5 days.Then,the drug-containing serums were collected and added to the cultured HSC-T6.HSC proliferation was detected using MTT colorimetric assay.Collagen （Col） Ⅰ,Col Ⅲ and α-smooth muscle actin （SMA） of culture supernatant were detected by ELISA.The expression of transforming growth factor （TGF）-β31 in HSC was measured by immunocytochemistry staining,and the expressions of Gremlin and BMP7 were determined by RT-PCR and Western blot,respectively.Result：Compared with the control group,the HSC proliferation was significantly inhibited （P 〈 0.05）,the concentrations of Col Ⅰ,Col Ⅲ,α-SMA and the expression levels of TGF-β1 were decreased （P 〈0.01,P 〈0.05）,the mRNA and protein expressions of Gremlin were decreased （P 〈 0.01）,and the mRNA and protein expression of BMP7 were elevated （P 〈 0.01） in the lowand high-dose DSHX serum groups.The above results in DSHX serum groups were the same as the positive control serum group.Conclusion：DSHX has potential effect in treating hepatic fibrosis in rats.It could inhibit the proliferation and activation of HSC and reduce the synthesis of extra cellular matrix,which are partly achieved by down-regulating gremlin and up-regulating BMP7.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2015年第2期182-186,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
贵州省科技计划课题(黔科合SZ字[2008]3049)
