喘可治注射液对人胚胎肺成纤维细胞转化因子及相关蛋白的影响

Effect of Chuankezhi Injection on Transfer Factors and Related Proteins of Human Embryonic Lung Fibroblasts

摘要: 目的：通过喘可治注射液(CKZ)干预人胚胎肺成纤维细胞(Wi-38)，测定β转化生长因子(TGF-β)、1型胶原纤维蛋白(collagen1)和α-平滑肌肌动蛋白(α-SMA)，探讨CKZ在抑制细胞间质转化方面的作用机制。方法：将体外培养的Wi-38细胞随机分为6组：① 对照组，单独的DMEM培养；② 5 ng/mL TGF-β组，加入浓度为5 ng/mL的TGF-β孵育；③ 10 ng/mL TGF-β组，加入浓度为10 ng/mL的TGF-β孵育；④10 ul/mL喘可治注射液(CKZ)组，加入浓度为10 ul/mL CKZ的孵育；⑤ 20 ul/mL CKZ组，加入浓度为20 ul/mL CKZ的孵育；⑥ 10 ng/mL TGF-β + 20 ul/mL CKZ组，同时加入浓度10 ng/mL TGF-β和20 ul/mL CKZ孵育。采用实时荧光定量(qPCR)的方法检测孵育24 h、48 h、72 h的TGF-β、collagen 1、α-SMA的表达水平变化。结果：加入TGF-β处理组细胞的α-SMA、Collagen 1的mRNA水平升高、TGF-β的mRNA水平降低，与对照组相比，其差异均具有统计学意义(P < 0.05)；加入CKZ处理组细胞的α-SMA、TGF-β、Collagen 1的mRNA水平均降低，与对照组相比，其差异具有统计学意义(P < 0.05)；同时加入TGF-β和CKZ处理组细胞的α-SMA、Collagen 1的mRNA水平均显著降低，与TGF-β处理组相比，其差异具有统计学意义(P < 0.05)。结论：TGF-β、collagen 1、α-SMA参与了人胚胎肺成纤维细胞间质转化的病理进程，CKZ对这一进程起到了抑制作用，CKZ具有一定的抑制气道上皮间质转化的作用。

Abstract: Objective: To investigate the effect of Chuankezhi injection (CKZ) on human embryonic lung fibro-blasts (WI-38), and to explore the mechanism of CKZ in inhibiting the mesenchymal transition by measuring the levels of TGF-β, collagen 1 and α-smooth muscle actin (α-SMA). Method: The cultured Wi-38 cells were randomly divided into 6 groups: ① control group, DMEM incubatedalone; ② 5 ng/mL TGF-β group, incubated with 5 ng/mL TGF-β; ③ 10 ng/mL TGF-β group, incubated with 10 ng/ml TGF-β; ④ 10 ul/mL CKZ group, incubated with 10 ul/mL CKZ; ⑤ 20 ul/mL CKZ group, in-cubated with 20 ul/mL CKZ; ⑥ 10 ng/mL TGF-β + 20 ul/mL CKZ group, incubated with 10 ng/mL TGF-β and 20 ul/mL CKZ. The expression levels of TGF-β, collagen 1 and α-SMA were detected by qPCR at 24, 48 and 72 h. Result: Compared with the control group, the mRNA levels of α - SMA and collagen 1 were increased and the mRNA levels of TGF-β were decreased in the TGF-β treated group (P < 0.05). The mRNA levels of α-SMA, TGF-β and collagen 1 in CKZ treated cells were all significantly decreased (P < 0.05); Compared with the TGF-β treated group, the mRNA levels of α-SMA and colla-gen 1 were decreased in the both TGF-β and CKZ treated groups (P < 0.05). Conclusion: TGF-β, colla-gen 1 and α-SMA participate in the pathological process of mesenchymal transition of human em-bryonic lung fibroblasts. CKZ may inhibit this process, and CKZ may inhibit airway epithelial mes-enchymal transition to some extent.