基于TGF-β_(1)信号通路探讨清痹达金方对胶原诱导性关节炎-肺纤维化大鼠的影响  

Effect of Qingbi Dajin decoction on collagen induced arthritis with pulmonary fibrosis in rats based on TGF-β_(1) signal pathway

作　　者：杨静[1,2] 吴沅皞[1] YANG Jing;WU Yuanhao(The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine/National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学第一附属医院/国家中医针灸临床医学研究中心,天津300381 [2]天津中医药大学,天津301617

出　　处：《现代中西医结合杂志》2022年第21期2921-2928,3014,共9页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：天津市教委科研计划项目(2019ZD12)。

摘　　要：目的基于转化生长因子-β_(1)(TGF-β_(1))信号通路探讨清痹达金方对胶原诱导性关节炎-肺纤维化(CIA-PF)大鼠的干预作用及机制。方法将36只Wistar大鼠随机分为对照组、模型组、醋酸泼尼松组及清痹达金方高、中、低剂量组,每组6只。除对照组外,其余组大鼠通过气管注射博来霉素及尾根部注射牛Ⅱ型胶原方法建立CIA-PF模型。造模完成后,清痹达金方高、中、低剂量组分别给予10.8 g/(kg·d)、5.4 g/(kg·d)、2.7 g/(kg·d)清痹达金方灌胃,醋酸泼尼松组给予醋酸泼尼松1.35 mg/(kg·d)灌胃,对照组和模型组给予等体积的蒸馏水灌胃,连续灌胃14 d,期间观察大鼠一般情况、关节炎指数(AI)评分及足趾容积变化。末次灌胃结束后处死各组大鼠,ELISA法检测血清TGF-β_(1)水平,HE染色观察肺组织炎症程度,Masson染色观察肺组织纤维化程度,q-PCR法和Western blot法分别检测肺组织中TGF-β_(1)、E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)mRNA及蛋白表达情况。结果清痹达金方高、中剂量组大鼠AI评分和清痹达金方各剂量组大鼠足趾容积、血清TGF-β_(1)水平均明显低于模型组(P均<0.05);清痹达金方各剂量组大鼠肺泡结构完整,肺纤维化程度较模型组轻;清痹达金方各剂量组大鼠肺组织中TGF-β_(1)、α-SMA mRNA及蛋白相对表达量均明显低于模型组(P均<0.05),清痹达金方高、中剂量组大鼠肺组织中E-cadherin mRNA相对表达量及清痹达金方各剂量组大鼠肺组织中E-cadherin蛋白相对表达量均明显高于模型组(P均<0.05)。结论清痹达金方可以通过抑制TGF-β_(1)表达,增加其下游细胞因子E-cadherin表达和降低α-SMA表达,改善CIA-PF大鼠肺纤维化。Objective It is to investigate the therapeutic effect and intervention mechanism of Qingbi Dajin decoction in rats with collagen induced arthritis with pulmonary fibrosis(CIA-PF)based on transforming growth factor-β_(1)(TGF-β_(1))signaling pathway.Methods Thirty six Wistar rats were randomly divided into control group,model group,prednisone acetate group and high,medium and low dose groups of Qingbi Dajin decoction,with 6 rats in each group.The rats in all groups except for the control group were given bleomycin by intratracheal injection and bovine type II collagen by tail root injection to establish models of CIA-PF.After the modeling was completed,the high-dose,middle dose and low-dose groups of Qingbi Dajin decoction were respectively given 10.8 g/(kg·d),5.4 g/(kg·d)and 2.7 g/(kg·d)Qingbi Dajin decoction by gavage,the prednisone acetate group was given 1.35 mg/(kg·d)prednisone acetate by gavage,the control group and the model group were given equal volume of distilled water by gavage,all the groups were treated for 14 days.During this experiment,the changes in the general situation,the score of arthritis index(AI)and toe volume of the rats were observed.After the last gavage,the rats in each group were killed,the level of serum TGF-β_(1) was detected by ELISA,the degree of inflammation in lung tissue was observed by HE staining,the degree of fibrosis in lung tissue was observed by Masson staining,and mRNA and protein expression of TGF-β_(1),E-cadherin,α-smooth muscle actin(α-SMA)in lung tissue were detected by q-PCR and Western blot.Results The AI score of rats in high and middle dosage groups of Qingbi Dajin decoction,toe volume and the level of serum TGF-β_(1) of rats in each dosage group of Qingbi Dajin decoction were significantly lower than those of rats in the model group(all P<0.05);The alveolar structure of rats in each dose group of Qingbi Dajin decoction was complete,and the degree of pulmonary fibrosis was lighter than that in the model group;The relative expression of mRNA and protein o

关 键 词：胶原诱导性关节炎 肺纤维化 清痹达金方 转化生长因子-β_(1) E-钙黏蛋白 Α-平滑肌肌动蛋白 

分 类 号：R-332[医药卫生]