摘要
目的探讨山姜素对博来霉素所致肺纤维化(PF)小鼠的影响及作用机制。方法60只小鼠随机分为六组:对照组(NS组)、模型组(BLM组)、羧甲基纤维素钠组(CMC组)、低剂量山姜素组(Alp25组)、中剂量山姜素组(Alp50组)、高剂量山姜素组(Alp100组)。采用气管内滴入博来霉素(2U/kg)建立PF小鼠模型,从造模第二天开始治疗,隔天给药1次,共10次。21天后计算小鼠肺系数,测定肺组织中羟脯氨酸(HYP)的含量,HE染色和Masson染色观察肺部炎症和纤维化的程度,采用免疫组化检测TGF-β1、Smad2/3、Smad7表达,免疫荧光检测肺组织E-cad、α-SMA、Col-1表达,Western Blot检测Smad2/3、Smad7蛋白的表达,qRT-PCR检测TGF-β1 mRNA的表达。结果与NS组相比,BLM组、CMC组模型建立成功;与BLM组相比,Alp50组小鼠炎性细胞浸润明显减少、胶原纤维沉积明显减少、肺系数降低,肺组织中HYP含量减少(P<0.05),Col-1、α-SMA、Smad2/3、TGF-β1表达下降(均P<0.05),E-cad、Smad7表达增加(均P<0.05),肺纤维化明显改善。结论山姜素可缓解博来霉素所致的小鼠PF,其作用机制可能与抑制TGF-β1/Smad信号通路有关。
Objective To investigate the protective effect and mechanism of alpinetin(Alp)on bleomycin-induced pulmonary fibrosis(PF)in mice.Methods 60 mice were randomly divided into six groups:the control group(the NS group),the model group(the BLM group),the carboxymethylcellulose sodium group(the CMC group),the low-dose alpinetin group(the Alp25 group),the medium-dose alpinetin group(the Alp50 group)and the high-dose alpinetin group(the Alp100 group).The mice model of PF was established by intratracheal dripping of BLM(2U/kg).Starting from the second day of modeling,the drug was given once every other day for a total of 10 times.After 21 days,the lung coefficient of mice was calculated,the content of hydroxyproline(HYP)in lung tissue was measured,the degree of pulmonary inflammation and fibrosis was observed by HE staining and Masson staining,the expression of TGF-β1,Smad2/3 and Smad7 was detected by immunohistochemistry,the expression of E-cad,α-SMA and Col-1 in lung tissue was detected by immunofluorescence,and the expression of Smad2/3 and Smad7 protein was detected by western blotting.The expression of TGF-β1 mRNA was detected by reverse transcription-polymerase chain reaction(qRT-PCR).Results Compared with the NS group,the BLM group and the CMC group were successfully established the mice model of pulmonary fibrosis.Comparing with the BLM group,inflammatory cell infiltration,collagen fiber deposition,and lung coefficient decreased significantly in the Alp50 group,but the weight increased after treatment.The content of HYP in lung tissue decreased(P<0.05),the expression of Col-1,α-SMA,Smad2/3 and TGF-β1 decreased(all P<0.05),and the expression of E-cad and Smad7 increased(all P<0.05).The pulmonary fibrosis was significantly improved.Conclusion Alpinetin can relieve bleomycin-induced pulmonary fibrosis in mice,and its mechanism may be that it can inhibit the expression of TGF-β1/Smad signal pathway.
作者
刘宇
王倩
王荣丽
LIU Yu;WANG Qian;WANG Rong-li(Department of Respiratory and Critical Care Medicine,the Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646099,China)
出处
《临床肺科杂志》
2023年第6期898-904,共7页
Journal of Clinical Pulmonary Medicine
基金
四川省卫计委科研课题(No.17PJ367)。
