芪归方有效组分治疗特发性肺纤维化的实验研究(1)
摘要 目的：基于Smurf2串話TGF-β1/Smad信号通路研究芪归方有效组分对转化生长因子β1(TGF-β1)诱导人胚肺成纤维细胞MRC-5的影响及机制。方法：体外予以10 ng/mL TGF-β1培养MRC-5细胞，PCR仪检测α-SMA的mRNA水平判断模型是否成功。设立正常对照组、模型组(10 ng/mL TGF-β1处理)、泼尼松处理组、芪归方有效组分高剂量组、中剂量组、低剂量组，采用CCK-8法检测MRC-5的增殖情况，Western blot检测各组细胞Smad7、SnoN蛋白的表达水平，RT-PCR检测各组细胞Smurf2的mRNA水平。结果：与正常对照组比较，48 h模型组细胞增殖明显，α-SMA的mRNA水平增加(P<0.01)，造模成功。经10 ng/mL TGF-β1处理细胞MRC-5，与正常对照组比较，Smad7、SnoN蛋白水平显著降低，Smurf2蛋白mRNA水平升高(P<0.01或P<0.05)。不同浓度芪归方有效成分处理组细胞增殖降低，可逆转以上各指标的改变，呈一定的量效关系。结论：芪归方有效组分能抑制MRC-5细胞增殖，可通过减少MRC-5细胞中Smurf2所介导的Smad7、SnoN的泛素化，恢复Smad7、SnoN蛋白的表达，从而调节肺纤维化的发生发展。
关键词 芪归方有效组分;Smad泛素化调节因子2;Smad蛋白;核转录共抑制因子;特发性肺纤维化
Abstract Objective：Based on Smurf2 crosstalk TGF-β1/Smad signaling pathway to study the effect and mechanism of Qigui Fomula active components on transforming growth factor β1 (TGF-β1)-induced human embryo lung fibroblast MRC-5. Methods：MRC-5 was cultured with 10 ng/mL TGF-β1 in vitro， mRNA level of α-SMA was detected by PCR instrument to judge whether the model was successfully made. Normal control group， model group (10 ng/mL TGF-β1 treatment)， prednisone treatment group， high-dose group， middle-dose group and low-dose group of effective components of Qigui Fomula were set， CCK-8 method was used to detect the proliferation of MRC-5， Western blot was used to detect the expression level of Smad7 and SnoN protein in each group， and RT-PCR was used to detect the mRNA level of Smurf2 in each group. Results：Compared with the normal control group， the 48 h model group had significant cell proliferation， and the mRNA level of α-SMA increased (P<0.01)， and the modeling was successful. Compared with the normal control group， MRC-5 treated with 10 ng/mL TGF-β1 showed that Smad7 and SnoN protein levels were significantly reduced and Smurf2 protein mRNA levels were increased (P<0.01 or P<0.05). The cell proliferation of the Qigui Foluma group with different concentrations was reduced， which could reverse the changes of the above indicators and showed a certain dose-effect relationship. Conclusion：The effective components of Qigui Foluma can inhibit the proliferation of MRC-5 cells， and can reduce the ubiquitination of Smad7 and SnoN mediated by Smurf2 in MRC-5 cells， restore the expression of Smad7 and SnoN proteins， and thereby regulate the occurrence of pulmonary fibrosis development.
Keywords Effective Component of Qigui Foluma; Smad ubiquitination regulator 2; Smad protein;Nuclear transcription co-suppressor; Idiopathic pulmonary fibrosis, 百拇医药(彭艳芳 张莹雯 张亚兵)
关键词 芪归方有效组分;Smad泛素化调节因子2;Smad蛋白;核转录共抑制因子;特发性肺纤维化
Abstract Objective：Based on Smurf2 crosstalk TGF-β1/Smad signaling pathway to study the effect and mechanism of Qigui Fomula active components on transforming growth factor β1 (TGF-β1)-induced human embryo lung fibroblast MRC-5. Methods：MRC-5 was cultured with 10 ng/mL TGF-β1 in vitro， mRNA level of α-SMA was detected by PCR instrument to judge whether the model was successfully made. Normal control group， model group (10 ng/mL TGF-β1 treatment)， prednisone treatment group， high-dose group， middle-dose group and low-dose group of effective components of Qigui Fomula were set， CCK-8 method was used to detect the proliferation of MRC-5， Western blot was used to detect the expression level of Smad7 and SnoN protein in each group， and RT-PCR was used to detect the mRNA level of Smurf2 in each group. Results：Compared with the normal control group， the 48 h model group had significant cell proliferation， and the mRNA level of α-SMA increased (P<0.01)， and the modeling was successful. Compared with the normal control group， MRC-5 treated with 10 ng/mL TGF-β1 showed that Smad7 and SnoN protein levels were significantly reduced and Smurf2 protein mRNA levels were increased (P<0.01 or P<0.05). The cell proliferation of the Qigui Foluma group with different concentrations was reduced， which could reverse the changes of the above indicators and showed a certain dose-effect relationship. Conclusion：The effective components of Qigui Foluma can inhibit the proliferation of MRC-5 cells， and can reduce the ubiquitination of Smad7 and SnoN mediated by Smurf2 in MRC-5 cells， restore the expression of Smad7 and SnoN proteins， and thereby regulate the occurrence of pulmonary fibrosis development.
Keywords Effective Component of Qigui Foluma; Smad ubiquitination regulator 2; Smad protein;Nuclear transcription co-suppressor; Idiopathic pulmonary fibrosis, 百拇医药(彭艳芳 张莹雯 张亚兵)
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