Study on the inhibitory effect of ligustrazine on the proliferation and collagen production of human embryonic lung fibroblast MRC-5 / 国际中医中药杂志

ABSTRACT

Objective:To study the effect of ligustrazine on the proliferation and collagen production of human embryonic lung fibroblasts MRC-5.Methods:MRC-5 was cultured and divided into control group, low-dose group, medium-dose group and high-dose group. The control group was treated with DMEM without drugs, while the low-dose group, medium-dose group and high-dose group were treated with DMEM with different doses of ligustrazine. After 24, 36 and 48 h treatment, the cell proliferation activity was detected by MTS assay; the cell cycle was detect by flow cytometry; the apoptosis was measured by TUNEL staining; Western blot was used to detect the expression of Bcl-2, Bax, CyclinD1, P27 protein, and ELISA was used to detect the levels of TGF-β1, Col-Ⅰ and Col-Ⅲ.Results:After 24, 36 and 48 h treatment, compared to the control group, the proliferation inhibitory rate of low-, medium-, and high-dose group increased significantly ( P<0.05). After 48 h treatment, compared to the control group, the G0/G1 phase proportion of cell cycle in different doses of ligustrazine group significantly increased, and the S phase proportion of cell cycle in different doses of ligustrazine group significantly decreased ( P<0.05). Compared to the control group, the apoptosis rate (6.59% ± 0.95%, 10.92% ± 2.25%, 16.58% ± 3.25% vs. 1.38% ± 0.34%) in different doses of ligustrazine group significantly increased ( P<0.05). Compared to the control group, the expression of Bcl-2 (0.79 ± 0.13, 0.52 ± 0.06, 0.31 ± 0.05 vs. 0.91 ± 0.15) and CyclinD1 (0.62 ± 0.08, 0.50 ± 0.06, 0.27 ± 0.04 vs. 0.83 ± 0.14) in different doses of ligustrazine group significantly decreased, while the expression of Bax (0.45 ± 0.07, 0.50 ± 0.06, 0.79 ± 0.13 vs. 0.32 ± 0.06) and p27 (0.39 ± 0.07, 0.75 ± 0.13, 0.96 ± 0.16 vs. 0.20 ± 0.05) significantly increased ( P<0.05). The content of TGF-β1, Col-Ⅰ and Col-Ⅲ in different doses of ligustrazine group were significantly decreased ( P<0.05). Conclusions:Ligustrazine can inhibit the proliferation and collagen production of human embryonic lung fibroblasts, which may be related to the induction of cell cycle arrest, regulation of proliferation and cell cycle related proteins expression.