摘要
目的研究当归补血总苷(TGDGBX)对转化生长因子β1(TGF-β1)诱导的人胚肺成纤维细胞(HLF-Ⅰ)转分化及胶原表达的调控作用。方法体外培养HLF-Ⅰ,根据不同的处理方法分为5组,A组:对照组;B组:加入TGF-β1(1ng/ml);C组:加入TGF-β1(1 ng/ml)+TGDGBX(0.03 mg/ml);D组:加入TGF-β1(1 ng/ml)+TGDGBX(0.09 mg/ml);E组:加入TGF-β1(1 ng/ml)+TGDGBX(0.27 mg/ml)。MMT法检测各组HLF-Ⅰ的增殖情况,羟脯氨酸消化法检测细胞上清液中羟脯氨酸(HYP)含量,ELISA法检测细胞上清液基质金属蛋白酶-9(MMP-9)的含量。半定量RT-PCR法检测Ⅰ型胶原(COL-Ⅰ)mRNA、Ⅲ胶原(COL-Ⅲ)mRNA、α-平滑肌肌动蛋白(α-SMA)mRNA以及MMP-9 mRNA的表达情况。结果与B组相比,C、D、E组明显抑制TGF-β1诱导的HLF-Ⅰ细胞增殖(P<0.05);降低细胞上清液中HYP的含量(P<0.05);减少HLF-Ⅰ的COL-ⅠmRNA、COL-ⅢmR-NA以及α-SMA mRNA的表达(P<0.05);增加细胞上清液中MMP-9的含量(P<0.05)。D和E组与B组相比,显著提高HLF-Ⅰ的MMP-9 mRNA表达水平(P<0.05,P<0.01)。结论 TGDGBX可抑制TGF-β1诱导的HLF-Ⅰ转分化和胶原表达,其抑制胶原表达的作用可能是通过增加MMP-9的表达来实现的。
Objective To investigate the mechanism of TGDGBX regulating expression of collagen in human fetal lung fibroblast（HLF-Ⅰ） triggered by transforming growth factor-β1（TGF-β1）.Methods Fibroblasts was cultured by α-MEM medium and was grouped according to treatment.Groups as follows,A： control;B： TGF-β1（1 ng/ml）;C：TGF-β1（1 ng/ml） ＋ TGDGBX（0.03 mg/ml）;D：TGF-β1（1 ng/ml） ＋TGDGBX（0.09 mg/ml）,E：TGF-β1（1 ng/ml）＋TGDGBX（0.27 mg/ml）.Proliferation of cells was measured by MTT assay.The changes of collagen-Ⅰ（COL-Ⅰ）,collagen-Ⅲ（COL-Ⅲ）,α-SMA and MMP-9 mRNA was detected by reverse transcription-polymerase chain reaction（RT-PCR） and the expression of MMP-9 in the supernatants were detected with ELISA.Results The cell multiplication of groupe TGDGBX was significantly higher than that in group TGF-β1（P0.05）;The collagen Ⅰ（COL-Ⅰ）,collagen Ⅲ（COL-Ⅲ） and α-SMA mRNA expression of fibroblasts in group TGDGBX were significantly lower than that in group TGF-β1（P0.05）,but the expression of MMP-9 mRNA in group TGDGBX was higher than that in group TGF-β1（P0.01）;TGDGBX could induce up-regulation of MMP-9 content of the supernatant.Conclusion TGDGBX can inhibite collagen expression by reducing HFL-I to MFB transdifferentiation and increase the expression of MMP-9 mRNA of HLF-Ⅰ.
出处
《安徽医科大学学报》
CAS
北大核心
2011年第10期1018-1022,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然基金青年科学资金资助项目(编号:30801535)
