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目的研究久效磷对体外培养的大鼠皮肤成纤维细胞生长及胶原Ⅰ和Ⅲ合成的影响。方法取4只出生3 d清洁级SD大鼠背部皮肤,以组织块法培养大鼠皮肤成纤维细胞,正常传代。取第4代成纤维细胞,按1.0×106个/瓶接种于25 cm2培养瓶中,当细胞至亚融合状态,分别加入0(对照)、0.01、0.1、1μg/L的久效磷溶液,培养24 h,镜下观察细胞生长状况,收集各组细胞。采用实时定量PCR(RT-PCR)检测大鼠皮肤成纤维细胞胶原Ⅰ和ⅢmRNA的表达;采用噻唑兰(MTT)比色法检测皮肤成纤维细胞的细胞活性。结果久效磷对体外培养的皮肤成纤维细胞有显著的抑制作用,且抑制率呈剂量依赖性;0.01、0.1、1μg/L久效磷染毒组大鼠皮肤成纤维细胞抑制率均高于对照组,差异有统计学意义(P<0.05或P<0.01)。且随着久效磷染毒剂量的升高,大鼠皮肤成纤维细胞胶原Ⅰ和胶原ⅢmRNA的表达水平呈下降趋势;与对照组相比,0.01、0.1、1μg/L久效磷染毒组大鼠皮肤成纤维细胞胶原Ⅰ和胶原ⅢmRNA的表达水平均下降,差异有统计学意义(P<0.05或P<0.01)。结论久效磷可抑制大鼠皮肤成纤维细胞的生长,降低大鼠皮肤成纤维细胞胶原Ⅰ和ⅢmRNA的表达。
Objective To study the effects of monocrotophos on the growth of rat skin fibroblasts and the synthesis of collagen Ⅰ and Ⅲ in vitro. Methods Four dorsal skin of SD rats of 3 days of age were born. The rat skin fibroblasts were cultured by tissue block method and passaged normally. The 4th generation of fibroblasts were inoculated into 25 cm2 culture flasks at a density of 1.0 × 106 per flask. When the cells were sub-fused, 0 (control), 0.01, 0.1, 1 μg / After cultured for 24 h, the growth of the cells was observed under microscope, and the cells in each group were collected. Real-time quantitative PCR (RT-PCR) was used to detect the mRNA expression of collagen Ⅰ and Ⅲ in rat skin fibroblasts. The cell viability of the skin fibroblasts was detected by MTT assay. Results Monocrotophos significantly inhibited skin fibroblasts cultured in vitro and the inhibitory rate was dose-dependent. The inhibitory rates of skin fibroblasts in 0.01, 0.1 and 1 μg / L monocrotophos groups were significantly higher than those in monocrotophos Control group, the difference was statistically significant (P <0.05 or P <0.01). And with the increase of monocrotophos dose, the expression of collagen Ⅰ and collagen Ⅲ mRNA in rat skin fibroblasts decreased. Compared with the control group, the concentrations of monocrotophos in the groups of 0.01, 0.1 and 1 μg / L monocrotophos The expression of collagen Ⅰ and collagen Ⅲ mRNA in rat skin fibroblasts decreased, with statistical significance (P <0.05 or P <0.01). Conclusion monocrotophos can inhibit the growth of rat skin fibroblasts and reduce the expression of collagen Ⅰ and ⅢmRNA in rat skin fibroblasts.