Xiao Hai, Xu Yi-ni, Luo Hong, Chen Yan, Zhang Yan-yan, Tao Ling, Jiang Yan, Shen Xiang-chun
Zhongguo Zhong Yao Za Zhi. 2015 Jun;40(11):2168-73.
To investigate the inhibitory effects of OMT on TGF-β1-induced CFBs proliferation, and then explore the mechanism.
The experiment was randomly divided into 6 groups as following: control group (serum free DMEM), model group (20 μg x L(-1) TGF-β1), OMT low dose group (1.89 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT medium dose group (3.78 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT high dose group (7.56 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), SB203580 group (p38MAPK blocking agent, 1 x 10(-5) mol x L(-1) + 20 μg x L(-1) TGF-β1). Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type I and collagen type III deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type I and collagen type III.
MTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-β1 (P < 0.01) α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type I and collagen type III by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-β1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-β1 (P < 0.01).
OMT can inhibit the CFBs proliferation induced by TGF-β1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.
研究氧化苦参碱（OMT）对转化生长因子-β1（TGF-β1）诱导的心脏成纤维细胞（CFBs）增殖的抑制作用，并探讨其机制。
实验随机分为6组：对照组（无血清DMEM）、模型组（20μg/L TGF-β1）、OMT低剂量组（1.89×10⁻⁴mol/L + 20μg/L TGF-β1）、OMT中剂量组（3.78×10⁻⁴mol/L + 20μg/L TGF-β1）、OMT高剂量组（7.56×10⁻⁴mol/L + 20μg/L TGF-β1）、SB203580组（p38丝裂原活化蛋白激酶（p38MAPK）阻断剂，1×10⁻⁵mol/L + 20μg/L TGF-β1）。采用免疫细胞化学方法鉴定CFBs的波形蛋白，以及肌成纤维细胞（myFBs）的α-平滑肌肌动蛋白（α-SMA）。采用MTT法检测OMT对CFBs增殖的抑制作用。苦味酸天狼星红染色分析Ⅰ型和Ⅲ型胶原沉积。蛋白质免疫印迹法检测p38MAPK、磷酸化p38MAPK（p-p38MAPK）、Ⅰ型胶原和Ⅲ型胶原的表达。
MTT结果显示，OMT显著抑制TGF-β1诱导的CFBs增殖（P < 0.01）。α-SMA免疫细胞化学实验表明，OMT可抑制CFBs增殖。通过蛋白质免疫印迹法和苦味酸天狼星红染色，OMT可显著减少Ⅰ型和Ⅲ型胶原的沉积。蛋白质免疫印迹结果显示，TGF-β1增强p38MAPK磷酸化，而OMT减弱TGF-β1诱导的p38MAPK磷酸化（P < 0.01）。
OMT可抑制TGF-β1诱导的CFBs增殖，其机制可能与抑制p38MAPK磷酸化有关。
