补阳还五汤对矽肺大鼠肺纤维化的影响及机制
|
摘要
目的观察补阳还五汤对矽肺大鼠纤维化的影响,并探讨其作用机制。方法将120只雄性Wistar大鼠分为对照组、模型组以及补阳还五汤高、中、剂量组,每组24只。模型组和补阳还五汤高、中、低剂量组采用一次性非暴露式二氧化硅气管滴注法制作矽肺模型,对照组气管内滴注生理盐水。造模成功后,补阳还五汤高、中、低剂量组分别给予18. 48、9. 24、4. 62 g/(kg·d)补阳还五汤灌胃,对照组和模型组均给予等量生理盐水灌胃,连续28d。于实验7、14、28 d分3批处死大鼠,采集肺组织,观察肺组织病理学改变,对肺泡炎程度和肺纤维化程度进行评分;采用免疫组化SP法检测肺组织中转化生长因子β1(TGF-β1)表达,Western blotting法检测肺组织中磷酸化细胞外信号调节激酶(p-ERK)、细胞外信号调节激酶1/2(ERK1/2)表达。结果与对照组相比,其他各组肺泡炎评分、肺纤维化评分在7、14、28 d时均升高(P均<0. 01);与模型组相比,补阳还五汤各剂量组肺泡炎评分、肺纤维化评分均下降(P均<0. 01)。与对照组相比,其他各组肺组织TGF-β1、p-ERK、ERK1/2表达均升高(P <0. 05或<0. 01);与模型组相比,补阳还五汤各剂量组肺组织TGF-β1、p-ERK、ERK1/2表达均降低(P <0. 05或<0. 01)。结论补阳还五汤可以有效降低矽肺大鼠的肺泡炎、抑制肺纤维化进程,该作用可能与调控TGF-β1/ERK信号通路、抑制其促增殖作用有关。
Objective To investigate the effect of Buyang Huanwu decoction on pulmonary fibrosis of silicosis rats and its mechanism. Methods Totally 120 male Wistar rats were divided into the control group,model group and high-,medium-and low-dose Buyang Huanwu decoction groups,with 24 rats in each group. The silicosis models in the model group and Buyang Huanwu decoction groups were made by one-time non-exposed silica intratracheal drip method,while the rats in the control group were given saline through intratracheal drip. After the successful modeling,the high-,medium-and lowdose Buyang Huanwu decoction groups were given intragastric administration of 18. 48,9. 24,and 4. 62 g/( kg·d) Buyang Huanwu decoction respectively,while the control group and model group were given the same amount of saline through intragastric administration for 28 consecutive days. On the 7 th,14 th and 28 th days of the experiment,rats were sacrificed in three batches. Lung tissues were collected to observe the pathological changes,and the degree of alveolitis and pulmonary fibrosis were scored. The expression of transforming growth factor-β1( TGF-β1) in the lung tissues was detected by immunohistochemical SP method. The expression of phosphorylated extracellular signal-regulated kinase( p-ERK) and extracellular signal-regulated kinase 1/2( ERK1/2) in the lung tissues were detected by Western blotting. Results Compared with the control group,the scores of alveolitis and pulmonary fibrosis in the other groups increased at 7,14 and 28 days( all P < 0. 01); compared with the model group,the scores of alveolitis and pulmonary fibrosis in the Buyang Huanwu decoction groups decreased( all P < 0. 01). Compared with the control group,the expression of TGF-β1,p-ERK and ERK1/2 in the lung tissues of other groups increased( P < 0. 05 or P < 0. 01); compared with the model group,the expression of TGF-β1,p-ERK and ERK1/2 in the lung tissues of Buyang Huanwu decoction groups decreased( P < 0. 05 or P < 0. 01).Conclusion Buyang Huanwu decoction can effectively reduce alveolitis in silicosis rats and inhibit the process of pulmonary fibrosis by regulating TGF-β1/ERK signaling pathway and inhibiting its proliferative effect.
Objective To investigate the effect of Buyang Huanwu decoction on pulmonary fibrosis of silicosis rats and its mechanism. Methods Totally 120 male Wistar rats were divided into the control group,model group and high-,medium-and low-dose Buyang Huanwu decoction groups,with 24 rats in each group. The silicosis models in the model group and Buyang Huanwu decoction groups were made by one-time non-exposed silica intratracheal drip method,while the rats in the control group were given saline through intratracheal drip. After the successful modeling,the high-,medium-and lowdose Buyang Huanwu decoction groups were given intragastric administration of 18. 48,9. 24,and 4. 62 g/( kg·d) Buyang Huanwu decoction respectively,while the control group and model group were given the same amount of saline through intragastric administration for 28 consecutive days. On the 7 th,14 th and 28 th days of the experiment,rats were sacrificed in three batches. Lung tissues were collected to observe the pathological changes,and the degree of alveolitis and pulmonary fibrosis were scored. The expression of transforming growth factor-β1( TGF-β1) in the lung tissues was detected by immunohistochemical SP method. The expression of phosphorylated extracellular signal-regulated kinase( p-ERK) and extracellular signal-regulated kinase 1/2( ERK1/2) in the lung tissues were detected by Western blotting. Results Compared with the control group,the scores of alveolitis and pulmonary fibrosis in the other groups increased at 7,14 and 28 days( all P < 0. 01); compared with the model group,the scores of alveolitis and pulmonary fibrosis in the Buyang Huanwu decoction groups decreased( all P < 0. 01). Compared with the control group,the expression of TGF-β1,p-ERK and ERK1/2 in the lung tissues of other groups increased( P < 0. 05 or P < 0. 01); compared with the model group,the expression of TGF-β1,p-ERK and ERK1/2 in the lung tissues of Buyang Huanwu decoction groups decreased( P < 0. 05 or P < 0. 01).Conclusion Buyang Huanwu decoction can effectively reduce alveolitis in silicosis rats and inhibit the process of pulmonary fibrosis by regulating TGF-β1/ERK signaling pathway and inhibiting its proliferative effect.
引文
[1]梁志欣,王平,陈良安.上皮细胞-间充质细胞转化及其在肺纤维化中的作用[J].中华内科杂志,2009,48(5):427-429.
[2]尚磊,李侠,宋占帅,等.补阳还五汤对矽尘所致肺纤维化大鼠血清PCⅢ和Ⅳ-C含量的影响[J].预防医学论坛,2018,24(1):16-19.
[3]姜岩,张娟,贾强,等.气管插管法建立矽肺大鼠模型方法的改进[J].现代预防医学,2017,44(22):4160-4164.
[4]Reilly MJ,Timmer SJ,Rosenman KD. The Burden of Silicosis inMichigan:1988-2016[J]. Ann Am Thorac Soc,2018,15(12):1404-1410.
[5]王焕强,李涛.尘肺病的定义与历史[J].中国职业医学,2017,44(4):485-493.
[6]Perri D,Cole DE,Friedman O,et al. Azathioprine and diffusealveolar hemorrhage:the pharmacogenetics of thiopurine methyl-transferase[J]. Eur Respir J,2007(30):1014-1017.
[7]黄成亮,李艳艳,范贤明,等.丹参联合川芎嗪对肺纤维化大鼠血清、支气管肺泡灌洗液中TNF-α和TGF-β1水平的影响[J].细胞与分子免疫学杂志,2013,29(7):673-676.
[8]邵笑,祝骥,赵峻峰,等.丹参酮ⅡA对大鼠肺纤维化细胞TGF-β1/Smads信号通路的影响[J].浙江中西医结合杂志,2016,26(5):414-417.
[9]Guo J,Gu N,Chen J,et al. Neutralization of interleukin-1 betaattenuates silica-induced lung inflammation and fibrosis in C57BL/6 mice[J]. Arch Toxicol,2013,87(11):1963-1973.
[10]Liu W,Wan J,Han JZ,et al. Antiflammin-1 attenuates bleomy-cin-induced pulmonary fibrosis in mice[J]. Respir Res,2013,101(14):1465-1475.
[11]李安,田琼,梁向艳,等.佤药灯台树对大鼠博莱霉素所致肺损伤早期作用的研究[J].科学技术与工程,2007,12(1):41-45.
[12]Park SH. Fine tuning and cross-talking of TGF-beta singal by in-hibitory smads[J]. J Biochem Mol Biol,2005,38(1):9-16.
[13]Flanders KC. Smad3 as a mediator of the fibrotic response[J]. IntJ Exp Pathol,2004,85(2):47-64.
[14]黄长海,杨敏,李惊自,等.结缔组织生长因子通过活化ERK1/2信号通路促成纤维细胞生成[J].中华医学杂志,2005,85:1322-1327.
[15]Kim CS,Kim JM,Nam SY,et al. Low-dose of ionizing radiationenhances cell proliferation via transient ERK1/2 and p38 activationin normal human lung fibroblasts[J]. J Radiat Res,2007,48(5):407-415.
[16]高建.当归补血总苦的研制及对实验性肺纤维化的干预作用和分子机制研究[D].合肥:安徽大学,2009.
[2]尚磊,李侠,宋占帅,等.补阳还五汤对矽尘所致肺纤维化大鼠血清PCⅢ和Ⅳ-C含量的影响[J].预防医学论坛,2018,24(1):16-19.
[3]姜岩,张娟,贾强,等.气管插管法建立矽肺大鼠模型方法的改进[J].现代预防医学,2017,44(22):4160-4164.
[4]Reilly MJ,Timmer SJ,Rosenman KD. The Burden of Silicosis inMichigan:1988-2016[J]. Ann Am Thorac Soc,2018,15(12):1404-1410.
[5]王焕强,李涛.尘肺病的定义与历史[J].中国职业医学,2017,44(4):485-493.
[6]Perri D,Cole DE,Friedman O,et al. Azathioprine and diffusealveolar hemorrhage:the pharmacogenetics of thiopurine methyl-transferase[J]. Eur Respir J,2007(30):1014-1017.
[7]黄成亮,李艳艳,范贤明,等.丹参联合川芎嗪对肺纤维化大鼠血清、支气管肺泡灌洗液中TNF-α和TGF-β1水平的影响[J].细胞与分子免疫学杂志,2013,29(7):673-676.
[8]邵笑,祝骥,赵峻峰,等.丹参酮ⅡA对大鼠肺纤维化细胞TGF-β1/Smads信号通路的影响[J].浙江中西医结合杂志,2016,26(5):414-417.
[9]Guo J,Gu N,Chen J,et al. Neutralization of interleukin-1 betaattenuates silica-induced lung inflammation and fibrosis in C57BL/6 mice[J]. Arch Toxicol,2013,87(11):1963-1973.
[10]Liu W,Wan J,Han JZ,et al. Antiflammin-1 attenuates bleomy-cin-induced pulmonary fibrosis in mice[J]. Respir Res,2013,101(14):1465-1475.
[11]李安,田琼,梁向艳,等.佤药灯台树对大鼠博莱霉素所致肺损伤早期作用的研究[J].科学技术与工程,2007,12(1):41-45.
[12]Park SH. Fine tuning and cross-talking of TGF-beta singal by in-hibitory smads[J]. J Biochem Mol Biol,2005,38(1):9-16.
[13]Flanders KC. Smad3 as a mediator of the fibrotic response[J]. IntJ Exp Pathol,2004,85(2):47-64.
[14]黄长海,杨敏,李惊自,等.结缔组织生长因子通过活化ERK1/2信号通路促成纤维细胞生成[J].中华医学杂志,2005,85:1322-1327.
[15]Kim CS,Kim JM,Nam SY,et al. Low-dose of ionizing radiationenhances cell proliferation via transient ERK1/2 and p38 activationin normal human lung fibroblasts[J]. J Radiat Res,2007,48(5):407-415.
[16]高建.当归补血总苦的研制及对实验性肺纤维化的干预作用和分子机制研究[D].合肥:安徽大学,2009.