天龙竭调控Notch信号通路干预肺纤维化大鼠的实验研究  

Experimental study for TianLongjie regulation on Pulmonary Fibrosis Rats by Notch sig-naling pathway

作　　者：罗婷 袁德政 李蕾 李天纲 袁嘉丽 付义 LUO Ting;YUAN De-zheng;LI Lei;LI Tian-gang;YUAN Jia-li;FU Yi(Yunnan University of Chinese Medicine,Kunming Yunnan,650500,China;Chengdu University of Tradi-tional Chinese Medicine,Chengdu Sichuan,610000,China;Yunnan Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine for Chronic Disease in Prevention and Treatment,Kunming,Yunnan 650500,China;Fourth Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning,110101,China;Third Affilicated Hospital of Yunnan University of Chinese Medecine,Kunming 650504,China)

机构地区：[1]云南中医药大学,云南昆明650500 [2]成都中医药大学,四川成都610000 [3]云南省中西医结合慢病防治重点实验室,云南昆明650500 [4]辽宁中医药大学附属第四医院,辽宁沈阳110101 [5]云南中医药大学第三附属医院,云南昆明650504

出　　处：《时珍国医国药》2024年第9期2092-2098,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81360581,82274374);春城科技领军人才项目(2022SCP006);中医联合重大项目[2019FF002(-007),2017FF004(-007)]。

摘　　要：目的观察天龙竭对博来霉素诱导大鼠肺纤维化的Notch信号通路的影响,探讨天龙竭对肺纤维化(Pulmonary Fi-brosis,PF)的作用机理。方法将60只雄性Wistar大鼠随机分为空白组、模型组、天龙竭低、中、高剂量组、吡非尼酮组,每组10只,空白组除外,其余采用气管滴注博莱霉素制备大鼠肺纤维化模型,造模14d后,分别予天龙竭低、中、高剂量灌胃0.51g·kg^(-1)·d^(-1);1.01g·kg^(-1)·d^(-1);2.02g·kg^(-1)·d^(-1),吡非尼酮灌胃(0.12mg·kg^(-1)·d^(-1)),空白组和模型组予以相同量的生理盐水,每日给药1次,给药28d。肺功能检测后,收集肺组织并称重,计算肺指数。通过HE和Masson染色观察大鼠肺组织病理改变;实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blotting)检测TGF-β、α-SMA、COL-I、COL-III、Hes1、DLL4、Notch1以及免疫组织化学法检测VEGF的表达情况。结果与空白组比较,模型组大鼠肺指数均升高,HE和Masson染色示肺泡结构被破坏,大量炎性细胞浸润,肺间质大量蓝色胶原纤维沉积,纤维化病灶明显,肺组织TGF-β、α-SMA、VEGF、COL-I/III、Hes1、DLL4、Notch1 mRNA和蛋白表达上调;与模型组比较,天龙竭干预组大鼠肺指数降低,HE和Masson染色示肺组织炎症细胞浸润、肺泡结构破坏和纤维化病灶明显减轻,肺组织TGF-β、α-SMA、VEGF、COL-I/III、Hes1、DLL4、Notch1 mRNA和蛋白表达下调。结论天龙竭能够通过下调Notch信号通路相关基因VEGF、Hes1、DLL4、Notch1的表达,减少肺纤维化指标TGF-β、α-SMA的表达,降低PF大鼠肺指数和减少肺组织中COL-I/III胶原沉积,可能是通过调控Notch信号通路来实现改善肺纤维化。Objective To observe the effect of TianLongjie on the Notch signaling pathway of bleomycin-induced pulmonary fi-brosis in rats,and to explore the mechanism of TianLongjie on pulmonary fibrosis(PF).Methods Wistar rats were replicated by endotracheal infusion of bleomycin(Bleomycin,BLM),and then which were randomly divided into model group(Model),Tianlong low dose group(TL),Tianlong medium dose group(TM),Tianlong high dose group(TH),and positive drug pirfenidone group(PFD),except the blank group(Control),was with 10 rats in each group.A rat pulmonary fibrosis model was prepared by tra-cheal instillation of bleomycin,and after 14 days of modeling,TianLongjie was given by low,medium and high doses of intragas-tric administration(0.51g·kg^(-1)·d^(-1);1.01g·kg^(-1)·d^(-1);2.02g·kg^(-1)·d^(-1)),pirfenidone was given by gavage(0.12mg·kg^(-1)·d^(-1)),and the blank group and the model group were given the same amount of normal saline,administered once a day for 28 days.After pulmonary function tests,lung tissue is collected and weighed to calculate the lung index.Hematoxylin-eosin staining(HE)and Masson staining were used to observe the pathological changes of rat lung tissue.The expression of TGF-β,α-SMA,COL-I,COL-III,Hes1,DLL4,Notch1 and immunohistochemistry was detected by real-time polymerase chain re-action(real-time PCR)and Western blotting.Results Compared with the control group,the lung index of the model group was increased,and the alveolar structure was destroyed by HE and Masson staining,a large number of inflammatory cells were infiltra-ted,blue collagen fibers were deposited in the lung interstitium,and the fibrotic lesions were obvious,the mRNA and protein ex-pressions of TGF-β,α-SMA,VEGF,COL-I,COL-III,Hes1,DLL4 and Notch1 were up-regulated in lung tissues.Compared with the model group,the lung index of the rats in the Tianlong exhaustion intervention group decreased,and HE and Masson staining showed that the inflammatory cell infiltration,alveolar structure destruction and fibrotic lesions in lun

关 键 词：天龙竭 NOTCH信号通路 肺纤维化 

分 类 号：R285.5[医药卫生—中药学]