养阴益气合剂对肺纤维化大鼠肺泡上皮间质转化的干预机制研究
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摘要
目的研究养阴益气合剂对博来霉素诱导的肺纤维化大鼠肺泡上皮间质转化的干预作用。方法将120只SPF级Wistar大鼠随机分为正常组,模型组,泼尼松组,养阴益气合剂低、中、高剂量组,采用气管滴注博来霉素的方法复制肺纤维化大鼠模型,造模后1 d各组给予相应药物灌胃,连续给药28 d。于给药第14天和28天分批处死大鼠,左肺行HE染色和Masson染色观察病理变化,右肺用RT-PCR法检测α-平滑肌肌动蛋白(α-SMA)、E-钙黏附蛋白(E-cadherin)、I型胶原(ColI)、层黏连蛋白(LN)mRNA的表达。结果与正常组相比,模型组α-SMA、Col I及LN mRNA表达均升高,E-cadherin表达下降(P<0.05)。各治疗组α-SMA、Col I及LN mRNA出现不同程度下降,E-cadherin mRNA表达上调(P<0.05)。第28天,与泼尼松组比较,养阴益气合剂高剂量组E-cadherin mRNA表达上调(P<0.05),养阴益气高、中、低剂量组LN mRNA表达下调(P<0.05)。结论养阴益气合剂能抑制博来霉素诱导的肺纤维化大鼠肺泡上皮间质转化,其机制可能与下调α-SMA、Col I、LN mRNA表达、上调E-cadherin mRNA表达有关。
Objective To study the interventional effect of Yangyin Yiqi Heji(Yin-nourishing Qireplenishing Mixture)to alveolus epithelial-mesenchymal transition in rats with bleomycin-induced pulmonary fibrosis.Methods Wistar rats in SPF class(n=120)were randomly divided into normal group,model group,prednisone group and low-dose,mid-dose and high-dose Yangyin Yiqi Heji groups.The rat model of pulmonary fibrosis was copied by using intratracheal instillation of bleomycin,and all groups were orally given corresponding medicinals for 28 d after modeling for 1 d.The rats were executed respectively on the 14th d and 28th d to collect lung samples for detection of left lung pathological changes by using HE and Masson staining.The mRNA expressions ofα-smooth muscle actin(α-SMA),E-cadherin,collagen I(Col I)and laminin(LN)were detected in rigth lung by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Results The mRNA expressions ofα-SMA,Col I and LN increased,and E-cadherin expressions decreased in model group compared with normal group(P<0.05).After treatment,the mRNA expressions ofα-SMA,Col I and LN decreased,and mRNA expression of E-cadherin increased in all treatment groups(P<0.05).On the 28 th d,mRNA expression of E-cadherin increased in high-dose Yangyin Yiqi Heji group compared with prednisone groups(P<0.05),and mRNA expression of LN deecresed in high-dose,mid-dose and low-dose Yangyin Yiqi Heji groups(P<0.05).Conclusion Yangyin Yiqi Heji can inhibit the epithelial-mesenchymal transition in rats with bleomycin-induced pulmonary fibrosis,and the mechanism may be related to down-regulation of genes ofα-SMA,Col I and LN and up-regulation of E-cadherin gene.
Objective To study the interventional effect of Yangyin Yiqi Heji(Yin-nourishing Qireplenishing Mixture)to alveolus epithelial-mesenchymal transition in rats with bleomycin-induced pulmonary fibrosis.Methods Wistar rats in SPF class(n=120)were randomly divided into normal group,model group,prednisone group and low-dose,mid-dose and high-dose Yangyin Yiqi Heji groups.The rat model of pulmonary fibrosis was copied by using intratracheal instillation of bleomycin,and all groups were orally given corresponding medicinals for 28 d after modeling for 1 d.The rats were executed respectively on the 14th d and 28th d to collect lung samples for detection of left lung pathological changes by using HE and Masson staining.The mRNA expressions ofα-smooth muscle actin(α-SMA),E-cadherin,collagen I(Col I)and laminin(LN)were detected in rigth lung by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Results The mRNA expressions ofα-SMA,Col I and LN increased,and E-cadherin expressions decreased in model group compared with normal group(P<0.05).After treatment,the mRNA expressions ofα-SMA,Col I and LN decreased,and mRNA expression of E-cadherin increased in all treatment groups(P<0.05).On the 28 th d,mRNA expression of E-cadherin increased in high-dose Yangyin Yiqi Heji group compared with prednisone groups(P<0.05),and mRNA expression of LN deecresed in high-dose,mid-dose and low-dose Yangyin Yiqi Heji groups(P<0.05).Conclusion Yangyin Yiqi Heji can inhibit the epithelial-mesenchymal transition in rats with bleomycin-induced pulmonary fibrosis,and the mechanism may be related to down-regulation of genes ofα-SMA,Col I and LN and up-regulation of E-cadherin gene.
引文
[1]Jolly MK,Ward C,Eapen MS,et al.Epithelial-mesenchymal transition,a spectrum of states:role in lung development,homeostasis and disease[J].Developmental Dynamics,2017,247(3):346-358.
[2]刘祉宁.上皮-间充质转化与肺纤维化的研究进展[J].齐齐哈尔医学院学报,2018,39(8):950-953.Liu ZN.Research progress in epithelial-mesenchymal transition and pulmonary fibrosis[J].Journal of Qiqihar Medical University,2018,39(8):950-953.
[3]Yamada A,Aki T,Unuma K,et al.Paraquat induces epithelial-mesenchymal transition-like cellular response resulting in fibrogenesis and the prevention of apoptosis in human pulmonary epithelial cells[J/OL].PLoS One,2015,10(3):e120192[2015-03-23]https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0120192.
[4]王洪,李霞,辛大永,等.养阴益气合剂对特发性肺间质纤维化气阴两虚证患者生活质量的影响[J].中医杂志,2016,57(9):767-770.Wang H,Li X,Xin DY,et al.Effect of Yangyin Yiqi Mixture on quality of life in patients of idiopathic pulmonary fibrosis with deficiency of both qi and yin syndrome[J].Journal of Traditional Chinese Medicine,2016,57(9):767-770.
[5]赵晨宇,祝勇,周济谱,等.养阴益气合剂对博来霉素所致肺纤维化大鼠的影响[J].中国中医药信息杂志,2015,22(9):63-67.Zhao CY,Zhu Y,Zhou JP,et al.Effects of Yangyin Yiqi Mixture on bleomycin-induced pulmonary fibrosis in rats[J].Chinese Journal of Information on TCM,2015,22(9):63-67.
[6]Kato S,Inui N,Hakamata A,et al.Changes in pulmonary endothelial cell properties during bleomycin-induced pulmonary fibrosis[J/OL].Respiratory Research,2018,19(1):127[2018-06-26].https://respiratoryresearch.biomedcentral.com/articles/10.11861a1293-018-0831-y.
[7]Bhatt T,Rizvi A,Batta SP,et al.Signaling and mechanical roles of E-cadherin[J].Cell Commun Adhes,2013,20(6):189-199.
[8]Kyung SY,Kim DY,Yoon JY,et al.Sulforaphane attenuates pulmonary fibrosis by inhibiting the epithelialmesenchymal transition[J].BMC Pharmacology&Toxicology,2018,19(1):13-23.
[9]Ji Y,Dou YN,Zhao QW,et al.Paeoniflorin suppresses TGF-βmediated epithelial-mesenchymal transition in pulmonary fibrosis through a Smad-dependent pathway[J].Acta Pharmacologica Sinica,2016,37(6):794-804.
[10]马跃文,佟振月,侯显明.肺纤维化过程中不同种类胶原异常沉积及分布规律的实验研究[J].中国医科大学学报,2009,38(6):407-409.Ma YW,Tong ZY,Hou XM.Abnormal deposition and distribution of different types of collagen in pulmonary fibrosis:an experimental study[J].Journal of China Medical University,2009,38(6):407-409.
[11]Hoozen BE,Grimmer KL,Marelich GP,et al.Early phase collagen synthesis in lungs of rats exposed to bleomycin[J].Toxicology,2000,147(1):1-13.
[12]Kuhn C,McDonald JA.The roles of the myofibroblast in idiopathic pulmonary fibrosis,ultrastructural and immunohistochemical features of sites of active extracellular matrix synthesis[J].Am J Patho,1991,138(5):1257-1265.
[13]刘哲,张晓梅,尹婷,等.肺纤方提取物干预肺纤维化模型大鼠肺微血管内皮细胞新生血管形成的体外研究[J].北京中医药大学学报,2015,38(4):233-235.Liu Z,Zhang XM,Yin T,et al.Intervention of Feixian Formula extract on pulmonary microvascular endothelial cell migration and angiogenesis in vitro in rat model of pulmo-nary fibrosis[J].Journal of Beijing University of Traditional Chinese Medicine,2015,38(4):233-235.
[14]王晨,王紫娟,张晓梅,等.肺纤方提取物对肺间质纤维化大鼠肺微血管VEGF、VEGFR2、PAI-1、VCAM-1 mRNA表达的影响[J].北京中医药大学学报,2017,40(2):143-147.Wang C,Wang ZJ,Zhang XM,et al.Effect of Feixian Fang extract on mRNA expression of VEGF,VEGFR2,PAI-1 and VCAM-1 in microvascular endothelial cells of rats with pulmonary fibrosis:an in vitro study[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(2):143-147.
[15]张伟,郑建,朱雪,等.益气类中药对博来霉素所致肺纤维化大鼠血管新生机制的影响[J].中医药信息,2014,31(4):85-88.Zhang W,Zhen J,Zhu X,et al.Influence of Chinese medicinal with qi-supplementing effect on angiogenesis mechanism in rats with bleomycin-induced pulmonary fibrosis[J].Information on Traditional Chinese Medicine,2014,31(4):85-88.
[16]王丽芹.养阴类中药对博来霉素致肺纤维化大鼠的干预作用及机制研究[D].济南:山东中医药大学,2014.Wang LQ.Research about the effect and mechanism of yin tonics on bleomycin-induced pulmonary fibrosis in rats[D].Jinan:Shandong University of Traditional Chinese Medicine,2014.
[2]刘祉宁.上皮-间充质转化与肺纤维化的研究进展[J].齐齐哈尔医学院学报,2018,39(8):950-953.Liu ZN.Research progress in epithelial-mesenchymal transition and pulmonary fibrosis[J].Journal of Qiqihar Medical University,2018,39(8):950-953.
[3]Yamada A,Aki T,Unuma K,et al.Paraquat induces epithelial-mesenchymal transition-like cellular response resulting in fibrogenesis and the prevention of apoptosis in human pulmonary epithelial cells[J/OL].PLoS One,2015,10(3):e120192[2015-03-23]https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0120192.
[4]王洪,李霞,辛大永,等.养阴益气合剂对特发性肺间质纤维化气阴两虚证患者生活质量的影响[J].中医杂志,2016,57(9):767-770.Wang H,Li X,Xin DY,et al.Effect of Yangyin Yiqi Mixture on quality of life in patients of idiopathic pulmonary fibrosis with deficiency of both qi and yin syndrome[J].Journal of Traditional Chinese Medicine,2016,57(9):767-770.
[5]赵晨宇,祝勇,周济谱,等.养阴益气合剂对博来霉素所致肺纤维化大鼠的影响[J].中国中医药信息杂志,2015,22(9):63-67.Zhao CY,Zhu Y,Zhou JP,et al.Effects of Yangyin Yiqi Mixture on bleomycin-induced pulmonary fibrosis in rats[J].Chinese Journal of Information on TCM,2015,22(9):63-67.
[6]Kato S,Inui N,Hakamata A,et al.Changes in pulmonary endothelial cell properties during bleomycin-induced pulmonary fibrosis[J/OL].Respiratory Research,2018,19(1):127[2018-06-26].https://respiratoryresearch.biomedcentral.com/articles/10.11861a1293-018-0831-y.
[7]Bhatt T,Rizvi A,Batta SP,et al.Signaling and mechanical roles of E-cadherin[J].Cell Commun Adhes,2013,20(6):189-199.
[8]Kyung SY,Kim DY,Yoon JY,et al.Sulforaphane attenuates pulmonary fibrosis by inhibiting the epithelialmesenchymal transition[J].BMC Pharmacology&Toxicology,2018,19(1):13-23.
[9]Ji Y,Dou YN,Zhao QW,et al.Paeoniflorin suppresses TGF-βmediated epithelial-mesenchymal transition in pulmonary fibrosis through a Smad-dependent pathway[J].Acta Pharmacologica Sinica,2016,37(6):794-804.
[10]马跃文,佟振月,侯显明.肺纤维化过程中不同种类胶原异常沉积及分布规律的实验研究[J].中国医科大学学报,2009,38(6):407-409.Ma YW,Tong ZY,Hou XM.Abnormal deposition and distribution of different types of collagen in pulmonary fibrosis:an experimental study[J].Journal of China Medical University,2009,38(6):407-409.
[11]Hoozen BE,Grimmer KL,Marelich GP,et al.Early phase collagen synthesis in lungs of rats exposed to bleomycin[J].Toxicology,2000,147(1):1-13.
[12]Kuhn C,McDonald JA.The roles of the myofibroblast in idiopathic pulmonary fibrosis,ultrastructural and immunohistochemical features of sites of active extracellular matrix synthesis[J].Am J Patho,1991,138(5):1257-1265.
[13]刘哲,张晓梅,尹婷,等.肺纤方提取物干预肺纤维化模型大鼠肺微血管内皮细胞新生血管形成的体外研究[J].北京中医药大学学报,2015,38(4):233-235.Liu Z,Zhang XM,Yin T,et al.Intervention of Feixian Formula extract on pulmonary microvascular endothelial cell migration and angiogenesis in vitro in rat model of pulmo-nary fibrosis[J].Journal of Beijing University of Traditional Chinese Medicine,2015,38(4):233-235.
[14]王晨,王紫娟,张晓梅,等.肺纤方提取物对肺间质纤维化大鼠肺微血管VEGF、VEGFR2、PAI-1、VCAM-1 mRNA表达的影响[J].北京中医药大学学报,2017,40(2):143-147.Wang C,Wang ZJ,Zhang XM,et al.Effect of Feixian Fang extract on mRNA expression of VEGF,VEGFR2,PAI-1 and VCAM-1 in microvascular endothelial cells of rats with pulmonary fibrosis:an in vitro study[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(2):143-147.
[15]张伟,郑建,朱雪,等.益气类中药对博来霉素所致肺纤维化大鼠血管新生机制的影响[J].中医药信息,2014,31(4):85-88.Zhang W,Zhen J,Zhu X,et al.Influence of Chinese medicinal with qi-supplementing effect on angiogenesis mechanism in rats with bleomycin-induced pulmonary fibrosis[J].Information on Traditional Chinese Medicine,2014,31(4):85-88.
[16]王丽芹.养阴类中药对博来霉素致肺纤维化大鼠的干预作用及机制研究[D].济南:山东中医药大学,2014.Wang LQ.Research about the effect and mechanism of yin tonics on bleomycin-induced pulmonary fibrosis in rats[D].Jinan:Shandong University of Traditional Chinese Medicine,2014.