补肺清肝化瘀方对急性放射性肺炎大鼠巨噬细胞自噬活化的影响  

Effect of Bufei Qinggan Huayu Decoction on Autophagy Activation of Macrophages in Rats with Acute Radiation Pneumonia

作　　者：王佩 许文婧[1] 马理海 张浩 孟令占[1] Wang Pei;Xu Wenjing;Ma Lihai;Zhang Hao;Meng Lingzhan(Chongqing Traditional Chinese Medicine Hospital,Chongqing 400021,China)

机构地区：[1]重庆市中医院,重庆400021

出　　处：《中国中医急症》2023年第12期2098-2102,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：重庆市自然科学基金面上项目(cstc2020jcyj-msxmX1001)。

摘　　要：目的观察补肺清肝化瘀方对急性放射性肺炎(ARP)大鼠巨噬细胞自噬活化的影响。方法采用X射线照射诱导ARP模型。检测大鼠体质量、肺系数;HE染色观察肺组织病理损伤;Masson染色观察胶原沉积情况;ELISA检测肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、白细胞介素-4(IL-4)、白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)水平及肺组织丙二醛(MDA)、8-羟基脱氧鸟苷(8-OHdG)含量;流式细胞检测M1(CD68+CD86)和M2(CD68+CD206+)巨噬细胞比值及活性氧(ROS)含量;Western blotting检测肺组织p62、LC3Ⅰ/Ⅱ蛋白表达;免疫荧光双染(标记LC3B和CD68)观察巨噬细胞自噬情况。结果与对照组相比,模型组体质量第9天开始下降(P<0.05),肺系数升高(P<0.001),肺组织病理损伤严重,胶原沉积增加(P<0.001),促炎细胞因子IL-1β、IL-6、TNF-α、ROS、MDA、8-OHdG(P<0.01或P<0.001)和p62水平增加,抗炎细胞因子IL-4、IL-10、TGF-β水平、LC3B/CD68共表达、M2/M1和LC3-Ⅱ/LC3-Ⅰ比值降低(P<0.01或P<0.001)。补肺清肝化瘀方治疗后,大鼠体质量增加(P<0.05),肺系数降低(P<0.05),病理损伤有所改善,胶原沉积降低(P<0.001),促炎细胞因子水平下调,抗炎细胞因子水平上调,M2/M1比值增加(P<0.05),ROS、MDA和8-OHdG水平降低(P<0.05),p62蛋白表达、LC3B/CD68共表达和LC3-Ⅱ/LC3-Ⅰ比值增加(P<0.05或P<0.01或P<0.001)。结论补肺清肝化瘀方通过抑制M1极化,缓解氧化应激,激活ARP大鼠自噬。Objective:To investigate the effect of Bufei Qinggan Huayu Decoction on macrophage autophagy ac⁃tivation in acute radiation pneumonia(ARP)rats.Methods:The ARP model was induced by X-ray irradiation.The body weight and lung coefficient of rats were detected.HE staining was used to observe the pathological inju⁃ry of lung tissue.Masson staining was used to observe collagen deposition.The levels of TNF-α,IL-6,IL-1β,IL-4,IL-10 and TGF-βin alveolar lavage fluid and the content of MDA and 8-OHdG in lung tissue were detected by ELISA.The ratio of M1(CD68+CD86)and M2(CD68+CD206+)macrophages and ROS level were deter⁃mined by flow cytometry.The protein expression of p62 and LC3Ⅰ/Ⅱin lung tissue were detected by Western blotting.Immunofluorescence double staining(labeled LC3B and CD68)was used to observe the autophagy of macrophage.Results:Compared with the control group,the body weight decreased from day 9(P<0.05),and in⁃creased lung coefficient(P<0.001),serious pathological injury of lung tissue and increased collagen deposition(P<0.001),and levels of pro-inflammatory cytokines IL-1β,IL-6 and TNF-α,ROS,MDA and 8-OHdG(P<0.01 or P<0.001)and p62 increased,and level of anti-inflammatory IL-4,IL-10,TGF-β(P<0.01),LC3B/CD68 co-expression,ratio of M2/M1 and LC3-Ⅱ/LC3-Ⅰ(P<0.01 or P<0.001)decreased in model group.After treat⁃ment with Bufei Qinggan Huayu Decoction,the body weight increased(P<0.05),lung coefficient decreased(P<0.05),pathological injury was improved,collagen deposition decreased(P<0.001),pro-inflammatory cytokine lev⁃els were down-regulated,anti-inflammatory cytokine levels were up-regulated,M2/M1 ratio increased(P<0.05),ROS,MDA and 8-OHdG levels decreased(P<0.05)and p62 protein expression,and LC3B/CD68 co-expression and LC3-Ⅱ/LC3-Ⅰratio increased(P<0.05 or P<0.01 or P<0.001).Conclusion:Bufei Qinggan Huayu Decoc⁃tion can inhibit M1 polarization,relieve oxidative stress and activate autophagy in ARP rats.

关 键 词：急性放射性肺炎 补肺清肝化瘀方 巨噬细胞 自噬 大鼠 

分 类 号：R285.5[医药卫生—中药学]