大黄_虫丸对博来霉素致肺纤...ROCK信号通路表达的影响_蒋锋利.pdf

中华中医药杂志(原中国医药学报)2023年2月第38卷第2期 CJTCMP,February 2023,Vol.38,No.2 810 mediated NF-kappa B activation by targeting TAB2 and TAB3 for degradation.Nat Immunol,2008,9(4):369-37718 Zhang P,Cao Y,Chen S,et al.Combination of vinpocetine and dexamethasone alleviates cognitive impairment in nasopharyngeal carcinoma patients following radiation injury.Pharmacology,2021,106(1-2):37-4419 Wang S,Astsaturov I A,Bingham C A,et al.Effective antibody therapy induces host-protective antitumor immunity that is augmented by TLR4 agonist treatment.Cancer Immunol Immunother,2012,61(1):49-6120 Apetoh L,Ghiringhelli F,Tesniere A,et al.Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy.Nat Med,2007,13(9):1050-105921 Na S,Duan X,Wang R,et al.Chronic neuroinflammation induced by lipopolysaccharide injection into the third ventricle induces behavioral changes.J Mol Neurosci,2021,71(6):1306-131922 Baker R G,Hayden M S,Ghosh S.NF-kappaB,inflammation,and metabolic disease.Cell Metab,2011,13(1):11-2223 Lawrence T.The nuclear factor NF-kappaB pathway in inflammation.Cold Spring Harb Perspect Biol,2009,1(6):a165124 Liu J,Jiang M,Deng S,et al.miR-93-5p-Containing exosomes treatment attenuates acute myocardial infarction-induced myocardial damage.Mol Ther Nucleic Acids,2018,11:103-11525 Tang B,Xuan L,Tang M,et al.miR-93-3p alleviates lipopolysaccharide-induced inflammation and apoptosis in H9c2 cardiomyocytes by inhibiting toll-like receptor 4.Pathol Res Pract,2018,214(10):1686-169326 Shang Y,Dai S,Chen X,et al.MicroRNA-93 regulates the neurological function,cerebral edema and neuronal apoptosis of rats with intracerebral hemorrhage through TLR4/NF-kappaB signaling pathway.Cell Cycle,2019,18(22):3160-317627 Chen X,Deng T,Huo T,et al.MiR-140-5p/TLR4/NF-kappaB signaling pathway:Crucial role in inflammatory response in 16HBE cells induced by dust fall PM2.5.Ecotoxicol Environ Saf,2021,208:11141428 Liu H,Mao Z,Zhu J,et al.MiR-140-5p inhibits oxidized low-density lipoprotein-induced oxidative stress and cell apoptosis via targeting toll-like receptor 4.Gene Ther,2021,28(7-8):413-42129 Song W,Wang T,Shi B,et al.Neuroprotective effects of microRNA-140-5p on ischemic stroke in mice via regulation of the TLR4/NF-kappaB axis.Brain Res Bull,2021,168:8-16（收稿日期：2021年8月11日）大黄虫丸对博来霉素致肺纤维化大鼠 Rho/ROCK信号通路表达的影响蒋锋利，黄茂，史雨宸，蔡松，任士杰，孙文丽，张晶，张立山（北京中医药大学东直门医院呼吸科，北京 100700）摘要：目的：观察大黄虫丸对博来霉素致肺纤维化大鼠Rho/ROCK信号通路表达情况的影响，探讨其防治特发性肺纤维化（IPF）的作用机制。方法：将80只健康雄性SD大鼠随机分为空白对照组（Control组）、模型组（Model组）、吡非尼酮组（PFD组）、大黄虫丸大剂量组（DHZC-H组）、大黄虫丸中剂量组（DHZC-M组）、大黄虫丸小剂量组（DHZC-L组）。采用一次性气管内滴入博来霉素（5 mg/kg）复制肺纤维化大鼠模型；给药28 d后处死大鼠，行ELISA法检测TGF-1、TNF-含量；RT-PCR法检测ROCK1、-SMA mRNA表达；Western blot法检测p-RhoA、ROCK1、p-MLC蛋白表达。结果：与Model组比较，各治疗组TGF-1浓度和DHZC-L组TNF-浓度显著降低（P0.01，P0.05）；DHZC-L组TGF-1浓度显著低于PFD组（P0.05）。与Model组和PFD组比较，DHZC-M组和DHZC-L组ROCK1、-SMA mRNA相对表达量均显著降低（P0.01，P0.05）。与Model组比较，大黄虫丸各剂量组p-RhoA蛋白相对表达量以及各治疗组ROCK1、p-MLC蛋白相对表达量显著降低（P0.05，P0.01）；且DHZC-L组p-RhoA、ROCK1、p-MLC蛋白相对表达量显著低于PFD组（P0.01）。结论：大黄虫丸可通过下调TGF-1介导的RhoA/ROCK1信号通路及减少-SMA mRNA及TNF-表达，起到抗炎抗纤维化作用。研究报告通信作者：张立山，北京市东城区海运仓5号北京中医药大学东直门医院，邮编：100700，电话：010-84013357，E-mail：内文2.indd 8102023/2/28 10:36:06中华中医药杂志(原中国医药学报)2023年2月第38卷第2期 CJTCMP,February 2023,Vol.38,No.2 811 关键词：大黄虫丸；干血；特发性肺纤维化；-SMA；TGF-1；RhoA/ROCK1；肺络基金资助：国家自然科学基金项目（No.81673904），北京中医药大学东直门医院2020科技创新专项（No.DZMKJCX-2020-025）Effects of Dahuang Zhechong Pills on the expression of Rho/ROCK signal pathway in Bleomycin-induced pulmonary fibrosis in ratsJIANG Feng-li,HUANG Mao,SHI Yu-chen,CAI Song,REN Shi-jie,SUN Wen-li,ZHANG Jing,ZHANG Li-shan(Department of Respiratory Medicine,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)Abstract:To observe the effects of Dahuang Zhechong Pills(DHZC)on the expression of Rho/ROCK signaling pathway in bleomycin-induced pulmonary fibrosis rats,and to explore its mechanism of prevention and treatment on idiopathic pulmonary fibrosis(IPF).Methods:Eighty healthy male SD rats were randomly divided into a blank control group(Control group),a model group(Model group),a pirfenidone group(PFD group),a large-dose group of DHZC(DHZC-H group),a medium-dose group of DHZC(DHZC-M group),a small-dose group of DHZC(DHZC-L group).Pulmonary fibrosis rat model was replicated with a one-time instillation of bleomycin(5 mg/kg)into the trachea;the rats were sacrificed 28 d after the administration,the TGF-1 and TNF-content was detected by the ELISA method;RT-PCR method was used to detect ROCK1 and-SMA mRNA expression;Western blot method was used to detect p-RhoA,ROCK1 and MLCP protein expression.Results:Compared with Model group,TGF-1 concentration in treatment groups and TNF-concentration in DHZC-L group were significantly decreased(P0.01,P0.05);The concentration of TGF-1 in DHZC-L group was significantly lower than that in PFD group(P0.05).Compared with Model group and PFD group,the mRNA relative expression levels of ROCK1 and-SMA in DHZC-M and DHZC-L groups were significantly decreased(P0.01,P0.05).Com