麦门冬汤对肺纤维化大鼠肺功能及内质网应激作用的影响
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摘要
目的探讨麦门冬汤改善博来霉素所致肺纤维化模型大鼠肺功能及内质网应激(ERS)作用的相关机制。方法健康雄性SD大鼠随机分为正常组、模型组、麦门冬汤1组(麦门冬与大枣质量比4∶1)、麦门冬汤2组(麦门冬与大枣质量比20∶1),经气管插管雾化注射博来霉素建立肺纤维化大鼠模型。造模后第14天每日早晚2次灌胃麦门冬汤,连续给药20 d。观察大鼠一般情况,通过HE和Masson染色观察肺组织的病理变化,检测大鼠用力肺活量(FVC)、0.4秒率(FEV0.4/FVC)、质量FVC、吸气阻力(Ri)、呼气阻力(Re)、肺顺应性(Cydn),并用Western blot和免疫组织化学染色分析肺组织中葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)的表达情况。结果与正常组相比,模型组FVC降低、0.4秒率升高、质量FVC下降、Cydn降低;与模型组相比,麦门冬汤1组和麦门冬汤2组FVC升高、0.4秒率降低,麦门冬汤2组质量FVC回升,麦门冬汤1组和麦门冬汤2组Cydn升高。病理可见模型组大鼠的肺组织实变区内慢性炎性细胞浸润,大量胶原沉积,高表达的GRP78和CHOP蛋白主要定位于空泡化的Ⅱ型肺泡上皮细胞(AECⅡs)。麦门冬汤2组大鼠炎症缓解,胶原沉积减少,肺组织中GRP78和CHOP蛋白的表达均减少。结论麦门冬汤能够明显改善大鼠肺功能,减少肺间质胶原沉积,可能与调节纤维化肺组织实变区内AECⅡs的GRP78和CHOP蛋白表达,缓解ERS压力,恢复AECⅡs的正常功能有关。
Objective To investigate the relative mechanism of Maimendong Tang(Ophiopogon Decoction)in amelioration of lung function and endoplasmic reticulum stress(ERS)in rats with bleomycin-induced pulmonary fibrosis.Methods Male healthy SD rats were randomly divided into normal group,model group,Maimendong Tang group 1 and Maimendong Tang group 2.The rat model of pulmonary fibrosis was established by bleomycin pulverization via tracheal intubation.On the 14 t h d after modeling,Maimendong Tang was intragastrically given to Maimendong Tang group1 and Maimendong Tang group 2 twice a day for 20 d continuously.The general conditions were observed,and pathological changes of lung tissue were observed after HE and Masson staining.The forced vital capacity(FVC),0.4 second rate(FEV0.4/FVC),mass FVC,inspiratory resistance(Ri),expiratory resistance(Re)and dynamic pulmonary compliance(Cydn)were detected by using spirometer.The expressions of glucose regulated protein 78(GRP78)and C/EBP homologous protein(CHOP)protein in lung tissue were analyzed by using Western blotting assay and immunohistochemistry staining technique.Results Compared with normal group,FVC,FVC/lung mass and Cydn decreased,and 0.4 second rate increased in model group.Compared with model group,FVC increased and 0.4 second rate decreased and Cydn increased in Maimendong Tang group 1 and Maimendong Tang group 2,and mass FVC increased in Maimendong Tang group 2.Pathological sections showed that there were chronic inflammatory cell infiltration in consolidation zone of lung tissue and massive collagen deposition,GRP78 and CHOP with high expressions were mainly positioned in vesicular type Ⅱ alveolar epithelial cells(AECⅡs).The inflammation was relieved,collagen deposition was decreased,protein expressions of GRP78 and CHOP were reduced and in Maimendong Tang group 2.Conclusion Maimendong Tang can significantly improve lung function and reduce lung interstitial collagen deposition,which may be related to regulation of protein expressions of GRP78 and CHOP in AECⅡs in consolidation zone of lung tissue,relief of ERS,and recovery of normal AECⅡs function.
Objective To investigate the relative mechanism of Maimendong Tang(Ophiopogon Decoction)in amelioration of lung function and endoplasmic reticulum stress(ERS)in rats with bleomycin-induced pulmonary fibrosis.Methods Male healthy SD rats were randomly divided into normal group,model group,Maimendong Tang group 1 and Maimendong Tang group 2.The rat model of pulmonary fibrosis was established by bleomycin pulverization via tracheal intubation.On the 14 t h d after modeling,Maimendong Tang was intragastrically given to Maimendong Tang group1 and Maimendong Tang group 2 twice a day for 20 d continuously.The general conditions were observed,and pathological changes of lung tissue were observed after HE and Masson staining.The forced vital capacity(FVC),0.4 second rate(FEV0.4/FVC),mass FVC,inspiratory resistance(Ri),expiratory resistance(Re)and dynamic pulmonary compliance(Cydn)were detected by using spirometer.The expressions of glucose regulated protein 78(GRP78)and C/EBP homologous protein(CHOP)protein in lung tissue were analyzed by using Western blotting assay and immunohistochemistry staining technique.Results Compared with normal group,FVC,FVC/lung mass and Cydn decreased,and 0.4 second rate increased in model group.Compared with model group,FVC increased and 0.4 second rate decreased and Cydn increased in Maimendong Tang group 1 and Maimendong Tang group 2,and mass FVC increased in Maimendong Tang group 2.Pathological sections showed that there were chronic inflammatory cell infiltration in consolidation zone of lung tissue and massive collagen deposition,GRP78 and CHOP with high expressions were mainly positioned in vesicular type Ⅱ alveolar epithelial cells(AECⅡs).The inflammation was relieved,collagen deposition was decreased,protein expressions of GRP78 and CHOP were reduced and in Maimendong Tang group 2.Conclusion Maimendong Tang can significantly improve lung function and reduce lung interstitial collagen deposition,which may be related to regulation of protein expressions of GRP78 and CHOP in AECⅡs in consolidation zone of lung tissue,relief of ERS,and recovery of normal AECⅡs function.
引文
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[14]吴逸园,杨业鹏,李载权.葡萄糖调节蛋白78研究进展[J].生理科学进展,2009,40(2):135-141.Wu YY,YANG YP,Li ZQ.Glucose regulated protein78kD[J].Progress in Physiological Sciences,2009,40(2):135-141.
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[17]Baek HA,Kim DS,Park H S,et al.Involvement of endoplasmic reticulum stress in myofibroblastic differentiation of lung fibroblasts[J].American Journal of Respiratory Cell&Molecular Biology,2012,46(6):731-739.
[2]康荻,李彧,朱曼,等.肺痿论治干预肺纤维化大鼠肺功能的实验研究[J].世界中医药,2017,12(2):390-393.Kang D,Li Y,Zhu M,et al.Experimental study on lung function of pulmonary fibrosis rat under lung withered theory[J].World Chinese Medicine,2017,12(2):390-393.
[3]Wynn TA.Integrating mechanisms of pulmonary fibrosis[J].Journal of Experimental Medicine,2011,208(7):1339-1350.
[4]Wolters PJ,Collard HR,Jones KD.Pathogenesis of idiopathic pulmonary fibrosis[J].Annual Review of Pathology,2014,9(1):157-179.
[5]Lawson WE,Roldan J,Cheng DS.Endoplasmic reticulum stress in alveolar epithelial cells is prominent in IPF:association with altered surfactant protein processing and herpesvirus infection[J].American Journal of Physiology,Lung Cellular and Molecular Physiology,2008,294(6):1119-1126.
[6]Lawson WE,Cheng DS,Degryse AL,et al.Endoplasmic reticulum stress enhances fibrotic remodeling in the lungs[J].Proceedings of the National Academy of Sciences,2011,108(26):10562-10567.
[7]李亚东.一种改善肺纤维化肺功能的药物及其制备方法:201810918015.4[P].2018-12-11.Li YD.The decoction for improving lung function of pulmonary fibrosis and its preparation method:201810918015.4[P].2018-12-11.
[8]朱曼,康荻,张慧,等.脾虚复合肺纤维化影响大鼠肺功能的初步研究[J].北京中医药大学学报,2017,40(2):130-134.Zhu M,Kang D,Zhang H,et al.Preliminary study of lung function affected by spleen deficiency pattern combined with pulmonary fibrosis of rats[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(02):130-134.
[9]晁恩祥,张纾难.肺痿再辨识[J].北京中医药大学学报,1997,20(5):14-15.Chao EX,Zhang SN.Re-identification atrophic lung disease[J].Journal of Beijing University of Traditional Chinese Medicine,1997,20(5):14-15.
[10]Walter P,Ron D.The unfolded protein response:from stress pathway to homeostatic regulation[J].Science,2011,334:1081-1086.
[11]Garg AD,Kaczmarek A,Krysko O,et al.ER stressinduced inflammation:does it aid or impede disease progression?[J].Trends in Molecular Medicine,2012,18(10):589-598.
[12]Zhao H,Wu QQ,Cao LF,et al.Correction:Melatonin inhibits endoplasmic reticulum stress and epithelialmesenchymal transition during bleomycin-induced pulmonary fibrosis in mice[J/OL].The Plos One Staff,2014,9(5):e97266[2015-03-25].http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0119381.
[13]Hsu HS,Liu CC,Lin JH.Involvement of ER stress,PI3K/AKT activation,and lung fibroblast proliferation in bleomycin-induced pulmonary fibrosis[J/OL].Scientific Reports,2017,7(1):14272[2017-10-27].http://www.nature.com/articles/s41598-017-14612-5.
[14]吴逸园,杨业鹏,李载权.葡萄糖调节蛋白78研究进展[J].生理科学进展,2009,40(2):135-141.Wu YY,YANG YP,Li ZQ.Glucose regulated protein78kD[J].Progress in Physiological Sciences,2009,40(2):135-141.
[15]Lee AS.The glucose-regulated proteins:stress induction and clinical applications[J].Trends in Biochemical Sciences,2001,26(8):504-510.
[16]Shimizu A,Kaira K,Yasuda M,et al.Clinical and Pathological Significance of ER Stress Marker(BiP/GRP78and PERK)Expression in Malignant Melanoma[J].Pathology&Oncology Research,2016,23(1):1-6.
[17]Baek HA,Kim DS,Park H S,et al.Involvement of endoplasmic reticulum stress in myofibroblastic differentiation of lung fibroblasts[J].American Journal of Respiratory Cell&Molecular Biology,2012,46(6):731-739.