化纤方通过上调外周及组织IFN-γ水平缓解放射性肺纤维化  

Hua Xian Fang alleviates radiation-induced pulmonary fibrosis by upregulating the level of IFN-γin blood and tissues

作　　者：陈君阳 邹坪金 方曾怡 公翠翠 殷婕 陈梅华 林冰[1] 郎锦义 Chen Junyang;Zou Pingjin;Fang Zengyi;Gong Cuicui;Yin Jie;Chen Meihua;Lin Bing;Lang Jinyi(Chengdu University of Traditional Chinese Medicine,Chengdu 610032,China;Department of Radiation Oncology,Radiation Oncology Key Laboratory of Sichuan Province,Sichuan Clinical Research Center for Cancer,Sichuan Cancer Hospital&Institute,Sichuan Cancer Center,Affiliated Cancer Hospital of University of Electronic Science and Technology of China,Chengdu 610041,China;Institute of Integrated Traditional Chinese and Western Medicine Cancer Research,Chengdu University of Traditional Chinese Medicine,Chengdu 610032,China;University of Electronic Science and Technology of China,Chengdu 611731,China)

机构地区：[1]成都中医药大学,成都610032 [2]放射肿瘤学四川省重点实验室,四川省肿瘤临床医学研究中心,四川省肿瘤医院·研究所,四川省癌症防治中心,电子科技大学附属肿瘤医院放疗中心,成都610041 [3]中西医结合肿瘤研究院,成都中医药大学,成都610032 [4]电子科技大学医学院,成都611731

出　　处：《中华放射肿瘤学杂志》2024年第6期554-561,共8页Chinese Journal of Radiation Oncology

基　　金：国家自然科学基金(81703070);四川省科技计划项目(2020YJ0446);四川省自然科学基金(2023NSFSC0708)。

摘　　要：目的:探究中药化纤方用于放射性肺纤维化的疗效及作用机制。方法:在体内实验中,36只6~8周龄雄性无特定病原(SPF)级C57BL/6小鼠被随机分为对照组、放射组(全肺17 Gy照射)、放射+化纤方组(全肺17 Gy照射+化纤方灌胃)。照射后16周,使用肺系数、HE染色和Masson染色评估肺泡炎症和肺纤维化程度,使用免疫组化染色检测肺组织α-平滑肌肌动蛋白(α-SMA)表达水平,使用实时荧光定量PCR(qPCR)法检测肺组织中γ干扰素(IFN-γ)的mRNA表达水平;同时,使用ELISA法测定血清及肺泡灌洗液中IFN-γ的含量。在体外实验中,使用IFN-γ刺激6 Gy照射后的成纤维细胞NIH/3T3,继续培养48 h后,通过qPCR、免疫荧光染色、蛋白质印记法(western blot,WB)检测α-SMA和Ⅰ型胶原蛋白(CollagenⅠ)转录及蛋白水平的表达。统计分析多组比较使用one-way ANOVA分析,组间多重比较采用Tukey检验。结果:与对照组相比,放射组的肺系数、肺泡炎评分、Ashcroft评分、肺组织α-SMA表达显著升高(P均<0.001)。相较于放射组,放射+化纤方组的上述指标均显著下降(P均<0.001)。qPCR检测结果显示放射+化纤方组IFN-γmRNA表达水平显著高于放射组(P=0.001);ELISA检测结果显示,与放射组相比,放射+化纤方组小鼠血清及肺泡灌洗液IFN-γ含量明显升高(P=0.032、0.037)。在体外实验中,放射后NIH/3T3细胞的α-SMA、Ⅰ型胶原蛋白的mRNA及蛋白表达显著升高,而在IFN-γ刺激后其表达明显降低。结论:化纤方能够有效缓解放射性肺纤维化,其机制与上调外周及组织IFN-γ,抑制成纤维细胞活化有关。Objective To investigate the therapeutic efficacy and underlying mechanisms of the traditional Chinese medicine formula"Hua Xian Fang"(HXF)in the treatment of radiation-induced pulmonary fibrosis(RIPF).Methods In vivo experiment,36 male specific pathogen free(SPF)-grade C57BL/6 mice aged 6-8 weeks were randomly divided into the control,irradiation(17 Gy thoracic irradiation),and irradiation+HXF groups(17 Gy thoracic irradiation+HXF).After 16 weeks,lung coefficient,HE staining and Masson staining were used to evaluate the degree of pulmonary inflammation and fibrosis.Immunohistochemistry was performed to measure the expression levels ofα-smooth muscle actin(α-SMA)in lung tissues.Quantitative real-time PCR(qPCR)was performed to detect the mRNA expression levels of interferon-γ(IFN-γ).Enzyme linked immunosorbent assay(ELISA)was conducted to determine the levels of IFN-γin serum and bronchoalveolar lavage fluid(BALF).During in vitro experiment,NIH/3T3 fibroblasts were stimulated with IFN-γafter 6 Gy irradiation,followed by 48 hours of culture.qPCR,immunofluorescence staining,and Western blot were used to assess the expression ofα-SMA and collagenⅠat the transcription and protein levels.One way ANOVA was used for multiple group comparisons,and Tukey test was used for inter group multiple comparisons.Results Compared to the control group,mice in the irradiation group showed significant increases in lung coefficient,Szapiel score,Ashcroft score,andα-SMA expression in lung tissues(all P<0.001).Compared to the irradiation group,the irradiation+HXF group exhibited significant decreases in the above indicators(all P<0.001).qPCR demonstrated that the mRNA expression levels of IFN-γwere significantly higher in the irradiation+HXF group than that in the irradiation group(P=0.001).ELISA results showed that the levels of IFN-γin serum and BALF were significantly elevated in the irradiation+HXF group compared to those in the irradiation group(P=0.032,0.037).In vitro experiment revealed that after irradiation,the expre

关 键 词：放射性肺纤维化 化纤方 Γ干扰素 成纤维细胞 Α-平滑肌肌动蛋白 

分 类 号：R563[医药卫生—呼吸系统]