益肺散结方激活自噬缓解COPD-PIF大鼠肺纤维化的实验研究

Experimental Research of Yifei Sanjie Recipe Activated Autophagy to Alleviate Pulmona-ry Fibrosis in COPD-PIF Rats

目的观察益肺散结方对慢性阻塞性肺疾病合并肺间质纤维化(Chronic Obstructive Pulmonary Disease with Pulmonary interstitial fibrosis,COPD-PIF)模型大鼠的作用效果并初步探讨其作用机制。方法分别用益肺散结方、地塞米松磷酸钠、益肺散结方叠加磷酸地塞米松干预模型大鼠,灌胃给药28天。于末次给药后一天取材。制作病理切片观察各组大鼠肺组织炎症及纤维化情况;酶联免疫吸附法检测各组大鼠肺组织匀浆中IL-8、TNF-α含量;碱水解法HYP试剂盒检测各组大鼠肺组织中羟脯胺酸含量;酶联免疫吸附法检测各组大鼠肺组织匀浆中的LN,HA含量;qPCR检测COPD-PIF大鼠肺组织中的自噬相关基因Beclin1、LC3B的mRNA表达水平。结果与模型组比较,益肺散结方干预组大鼠肺组织支气管周围炎性细胞浸润程度有所减轻;肺泡结构紊乱,肺泡壁变薄或断裂,肺泡腔扩大融合成肺大泡的症状有所缓解;益肺散结方干预组大鼠肺组织中IL-8、TNF-α、羟脯胺酸及LN含量均有不同程度的降低(P<0.05),HA含量有降低趋势但该差异没有统计学意义;益肺散结方干预组大鼠肺组织中自噬相关基因Beclin1、LC3B表达量均有所升高(P<0.05)。结论益肺散结方对COPD-PIF有一定的治疗作用,能显著改善模型大鼠肺组织损伤,减轻肺纤维化程度。其作用机制可能与激活细胞自噬有关。

Objective To observe the effect of Yifei Sanjie Fang on rats with chronic obstructive pulmonary disease with Pulmona-ry Interstitial Fibrosis(COPD-PIF)and to explore its mechanism.Methods Rats were treated with Yifei Sanjie Recipe,Dexam-ethasone Sodium Phosphate,Yifei Sanjie Decoction and dexamethasone phosphate for 28 days.The material was taken one day af-ter the last administration.The pathological sections were used to observe the inflammation and fibrosis of lung tissue in each group.The contents of IL-8 and TNF-αin lung tissue homogenate of each group were detected by enzyme-linked immunosor-bent assay(ELISA).The alkaline hydrolysis method was used to detect the large group of HYP kit.The content of hydroxyproline in rat lung tissue was detected by enzyme-linked immunosorbent assay(ELISA).Determination of LN and HA in lung tissue ho-mogenate of rats in each group by enzyme-linked immunosorbent assay.The mRNA expression levels of autophagy-related genes Beclin1 and LC3B in lung tissues of COPD-PIF rats were detected by qPCR.Results Compared with the model group,the de-gree of inflammatory cell infiltration around the bronchi of the lung tissue of the rats in the Yifei Sanjie Recipe group was reduced;the alveolar structure was disordered,the alveolar wall became thin or broken,and the alveolar cavity expanded into the pulmona-ry bullae.Alleviation of IL-8,TNF-α,hydroxyproline and LN in lung tissue of Yifei Sanjie Recipe group were significantly de-creased(P<0.05),HA content decreased,but the difference was not statistical significance;the expression of autophagy-re-lated genes Beclin1 and LC3B in lung tissue of Yifei Sanjie Recipe group were increased(P<0.05).Conclusion Yifei Sanjie Recipe has a certain therapeutic effect on COPD-PIF,which can significantly improve lung tissue damage and reduce the degree of pulmonary fibrosis.Its mechanism of action may be related to activating autophagy.