目的 研究右归丸含药血清抑制香烟烟雾提取物（cigarette smoke extract，CSE）＋转化生长因子-β1（transforming growth factor-β1，TGF-β1）诱导的大鼠II型肺泡上皮细胞（RLE-6TN）上皮间质转化（epithelial-mesenchymal transition，EMT）的作用机制。方法 RLE-6TN细胞经5% CSE处理12 h，再给予10 mg/L TGF-β1处理72 h建立EMT模型，给予5%、10%、15%、20%右归丸含药血清进行干预，采用CCK-8法测定细胞活力；采用免疫荧光法检测瘦素（leptin，LEP）表达；采用Western blotting检测LEP、E-钙黏蛋白（E-cadherin）和α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）表达。构建100 ng/mL LEP干预RLE-6TN细胞模型，给予10%右归丸含药血清进行干预，采用CCK-8法测定细胞活力；采用划痕实验测定细胞迁移能力；采用免疫荧光法检测E-cadherin、α-SMA表达；采用qRT-PCR法检测E-cadherin、α-SMA表达；采用Western blotting检测E-cadherin、α-SMA及Janus激酶2（Janus kinase 2，JAK2）/信号传导及转录激活因子3（signal transducer and activator of transcription 3，STAT3）信号通路相关蛋白表达。结果 RLE-6TN细胞经5% CSE＋10 mg/L TGF-β1干预后，LEP、α-SMA表达显著升高（P＜0.01），E-cadherin表达显著降低（P＜0.01）；与模型组比较，给予右归丸含药血清干预后，LEP、α-SMA表达显著降低（P＜0.05、0.01），E-cadherin表达显著升高（P＜0.05、0.01）。RLE-6TN细胞经100 ng/mL LEP干预后，细胞迁移率显著降低（P＜0.01），α-SMA蛋白及mRNA的表达显著升高（P＜0.01），E-cadherin蛋白及mRNA的表达显著降低（P＜0.01），p-JAK2/JAK2、p-STAT3/STAT3表达显著升高（P＜0.05、0.01）；与模型组比较，给予10%右归丸含药血清干预后，细胞迁移率显著升高（P＜0.01），α-SMA蛋白及mRNA的表达显著降低（P＜0.05、0.01），E-cadherin蛋白及mRNA的表达显著升高（P＜0.05、0.01），p-JAK2/JAK2、p-STAT3/STAT3表达显著降低（P＜0.05、0.01）。结论 右归丸含药血清能够通过降低LEP表达，抑制JAK2/STAT3信号通路，从而抑制RLE-6TN细胞EMT进程。

Objective To study the mechanism of Yougui Pills (右归丸) containing serum on inhibiting epithelial-mesenchymal transition (EMT) induced by cigarette smoke extract (CSE) and transforming growth factor-β1 (TGF-β1) in rat type II alveolar epithelial cells (RLE-6TN). Methods RLE-6TN cells were treated with 5% CSE for 12 h, followed by treatment with 10 mg/L TGF-β1 for 72 h to establish an EMT model. 5%, 10%, 15% and 20% Yougui Pills containing serum was administered for intervention, and cell viability was measured using CCK-8 method. Immunofluorescence method was used to detect the expression of leptin (LEP). Western blotting was used to detect the expressions of LEP, E-cadherin and α-smooth muscle actin (α-SMA). A model of RLE-6TN cells treated with 100 ng/mL LEP was established, followed by treatment with 10% Yougui Pills containing serum, and cell viability was measured using CCK-8 method, and measure cell viability using CCK-8 method. Scratch test was used to determine cell migration ability. Immunofluorescence assay was used to detect the expressions of E-cadherin and α-SMA. qRT-PCR was used to detect the expressions of E-cadherin and α-SMA. Western blotting was used to detect the expressions of E-cadherin, α-SMA and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway related proteins. Results After intervention with 5% CSE + 10 mg/L TGF-β1 in RLE-6TN cells, the expressions of LEP and α-SMA were significantly increased (P < 0.01), while the expression of E-cadherin was significantly decreased (P < 0.01). Compared with model group, after intervention with Yougui Pills containing serum, the expressions of LEP and α-SMA were significantly reduced (P < 0.05, 0.01), and the expression of E-cadherin was significantly increased (P < 0.05, 0.01). After intervention with 100 ng/mL LEP, the cell migration rate of RLE-6TN cells was significantly reduced (P < 0.01), the expressions of α-SMA protein and mRNA were significantly increased (P < 0.01), the expressions of E-cadherin protein and mRNA were significantly reduced (P < 0.01), the expressions of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly increased (P < 0.05, 0.01). Compared with model group, after intervention with 10% Yougui Pills containing serum, the cell migration rate was significantly increased (P < 0.01), the expressions of α-SMA protein and mRNA were significantly decreased (P < 0.05, 0.01), the expressions of E-cadherin protein and mRNA were significantly increased (P < 0.05, 0.01), the expressions of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly decreased (P < 0.05, 0.01). Conclusion Yougui Pills serum containing can inhibit the EMT process of RLE-6TN cells by reducing LEP expression and inhibiting the JAK2/STAT3 signaling pathway.

R285.5

国家自然科学基金青年项目（82104825）