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[目的]探讨RGD短肽对二氧化硅(SiO2)刺激引起的肺成纤维细胞激活效应的抑制作用。[方法]用无血清的SiO2培养基刺激巨噬细胞(AM)24h,离心收集培养液上清。加入RGD使其终浓度为50μg/ml、100μg/ml、150μg/ml,分别刺激LF;阳性对照未加RGD;阴性对照用不含SiO2的AM上清培养肺成纤维细胞(LF)。RT-PCR检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ⅰ型胶原(collagenⅠ,COLⅠ)及Ⅲ型胶原(collagenⅢ,COLⅢ)的mRNA水平;ELISA法测定COLⅠ和PCⅢ(procollagenⅢ,PCⅢ)。[结果]RGD浓度为100μg/ml时α-SMA、COLⅠ、COLⅢ的转录水平为(2.79±0.23、3.76±0.31、2.01±0.41),低于阳性对照组(4.47±0.84,5.03±0.15,3.07±0.41),差异有统计学意义(P﹤0.01);浓度为150μg/ml时,培养上清中COLⅠ、COLⅢ和总蛋白的表达为(1.82±0.45)μg/L、(1.91±0.41)μg/L、(0.09±0.06)μg/ml低于阳性对照组(3.00±0.69)μg/L,(3.47±0.96)μg/L,(0.28±0.06)μg/ml,差异有统计学意义(P﹤0.05)。[结论]RGD抑制COLⅠ、COLⅢ和总蛋白的表达,说明其对矽尘诱导大鼠肺成纤维细胞的激活效应有抑制作用。
[Objective] To investigate the inhibitory effect of RGD short peptides on the activation of lung fibroblasts induced by silica (SiO2) stimulation. [Method] The macrophages (AM) were stimulated with serum - free SiO2 medium for 24 hours, and the supernatant of the medium was collected by centrifugation. Addition of RGD to final concentrations of 50μg / ml, 100μg / ml and 150μg / ml respectively stimulated LF; positive control did not add RGD; negative control cultured lung fibroblasts (LF) with SiO2-free AM supernatant. The mRNA levels of α-smooth muscle actin (α-SMA), collagen Ⅰ (COLⅠ) and collagen Ⅲ (COLⅢ) were detected by RT-PCR. procollagen III, PC III). [Results] The transcriptional levels of α-SMA, COLⅠand COLⅢ at the RGD concentration of 100μg / ml were (2.79 ± 0.23, 3.76 ± 0.31 and 2.01 ± 0.41), lower than the positive control group (4.47 ± 0.84, 5.03 ± 0.15, 3.07 ± 0.41) (P <0.01). At the concentration of 150μg / ml, the expression of COL Ⅰ, COL Ⅲ and total protein in culture supernatant were (1.82 ± 0.45) μg / L, (1.91 ± 0.41) μg (0.09 ± 0.06) μg / ml and (0.09 ± 0.06) μg / ml respectively, which was significantly lower than that of positive control group (3.00 ± 0.69) μg / L and <0.05). [Conclusion] RGD inhibits the expression of COL Ⅰ, COL Ⅲ and total protein, indicating that RGD can inhibit the activation of rat lung fibroblasts induced by silica dust.