桃红四物汤对COPD合并肺癌模型小鼠抗炎作用的实验研究

Anti-inflammatory Effect of Taohong Siwu Decoction on COPD Combined with Lung Cancer in Mice

目的 观察桃红四物汤对慢性阻塞性肺疾病(Chronic obstructive pulmonary disease, COPD)合并肺癌模型小鼠外周血中炎症相关因子IL-4、IFN-γ、IL-17、TGF-β含量的影响,探讨桃红四物汤对COPD合并肺癌模型小鼠的影响及是否有抗炎作用。方法 将C57/BL6小鼠随机分为:空白对照组、空白+THSW组、COPD+Lewis对照组、COPD+Lewis+THSW组,每组各20只。首先利用烟熏加LPS气道内滴注的方法对COPD+Lewis对照组、COPD+Lewis+THSW组小鼠建立小鼠COPD模型,确定COPD模型复制成功一周后对COPD模型小鼠复制原位肺癌模型,最终建立COPD合并肺癌模型。空白对照组、COPD+Lewis对照组两组不做其他处理,空白+THSW组、COPD+Lewis+THSW组两组每日进行灌胃给药,桃红四物汤剂量:4.5 ml/kg, 1次/d,连续2周。HE染色法观察小鼠肺组织病理形态;酶联免疫吸附法(Enzyme-linked immunosorbent assay, ELISA)检测各组小鼠血清中白细胞介素4(Interleukin-4,IL-4)、干扰素γ(Interferon-gamma, IFN-γ)、白细胞介素17(Interleukin-17,IL-17)、转化生长因子-β(Transforming growth factor-β,TGF-β)含量。结果 HE染色示空白对照组和空白+THSW组小鼠肺组织未见异常。COPD+Lewis组小鼠肺组织病理改变严重,镜下可见大量炎性细胞浸润,肺泡结构紊乱。COPD+Lewis+THSW组小鼠肺组织炎性细胞浸润明显减少,肺泡较为清晰。与空白对照组和空白+THSW组比较,COPD+Lewis组和COPD+Lewis+THSW组小鼠血清中白细胞介素4(Interleukin-4,IL-4)、干扰素γ(Interferon-gamma, IFN-γ)、白细胞介素17(Interleukin-17,IL-17)、转化生长因子-β(Transforming growth factor-β,TGF-β)含量及IFN-γ/IL-4、IL-17/TGF-β比值明显升高,差异有统计学意义(P<0.01);与COPD+Lewis组比较,COPD+Lewis+THSW组小鼠血清中IL-4、IL-17含量及IFN-γ/IL-4、IL-17/TGF-β比值明显降低,而IL-4、TGF-β含量明显升高,差异有统计学意义(P<0.01)。结论 桃红四物汤可以减轻COPD合并肺癌的炎症反应,其作用机制可能是其抑制Th1、Th17型细胞因子IFN-γ、Treg型细胞因子IL-4、TGF-β的分泌,纠正Th1/Th2和Th17/Treg失衡状态有关。

Objective To observe the effect of Taohong Siwu Decoction(THSW)on the content of inflammatory factors IL-4,IFN-γ,IL-17,and TGF-βin the peripheral blood of chronic obstructive pulmonary disease(COPD)combined with lung cancer model mice,explore the effect of THSW on COPD complicated with lung cancer in mice,and verify the anti-inflammatory effect of THSW.Methods C57/BL6 mice were randomly divided into a blank control group(group A),a blank+THSW group(group B),a COPD+Lewis control group(group C),and a COPD+Lewis+THSW group(group D),with 20 mice in each group.The COPD model was induced by fumigation and LPS intratracheal instilla-tion in mice of groups C and D.One week after the COPD model induction,the orthotopic lung cancer model was replicated in COPD model mice to establish a COPD combined with the lung cancer model.The mice in groups A and C did not re-ceive additional treatment,while those in groups B and D were treated with THSW at 4.5 mL/kg by gavage,once per day for two consecutive weeks.HE staining was used to observe the pathological morphology of lung tissues in mice.Enzyme-linked immunosorbent assay(ELISA)was used to detect the content of Interleukin-4(IL-4),Interferon-gamma(IFN-γ),Interleukin-17(IL-17),and Transforming growth factor-β(TGF-β).Results HE staining showed that there was no abnormality in the lung tissues of mice in groups A and B.The pathological changes in lung tissues in group C were serious,and a large number of inflammatory cell infiltration and alveolar structure disorder were seen under the microscope.In the group D,the infiltration of inflammatory cells in lung tissues was significantly reduced,and the alveoli were clear.Compared with groups A and B,groups C and D showed increased levels of IL-4,IFN-γ,IL-17,and TGF-β,as well as IFN-γ/IL-4 and IL-17/TGF-βin serum(P<0.01).Compared with the group C,the group D showed reduced IL-4,IL-17,IFN-γ/IL-4,and IL-17/TGF-β,and increased IL-4 and TGF-β(P<0.01).Conclusion THSW can reduce the inflammatory reaction of COPD with lung cancer,and the underlying mechanism may be attributed to THSW can inhibit Th1 and Th17 type cytokines IFN-γand IL-17 secretion,promote Th2,Treg type cytokines IL-4,TGF-βsecretion,correcting the imbalance of Th1/Th2 and Th17/Treg.