Effects of enema decoction for alleviating pant on inflammatory response and cellular autophagy in rats with chronic obstructive pulmonary disease via activating AMPK pathway
Objective It is to observe the enema decoction for alleviating pant on inflammatory response and autophagy in rats with chronic obstructive pulmonary disease (COPD) via AMP-activated protein kinase (AMPK) signal pathway.Methods Sixty SPF-grade SD rats were randomly divided into blank group,model group,dexamethasone group,and low,medium,and high dose groups of enema decoction for alleviating pant,with 10 rats in each group.The rats of all groups except for the blank group were induced with cigarette smoking plus tracheal injection of lipopolysaccharide to establish models of COPD.At the end of modeling,the blank group and model group were given 0.9％ sodium chloride solution 5 mL/kg by enema,the dexamethasone group was given dexamethasone suspension 0.09 mg/kg by gavage,and the low,medium,and high dose groups of enema decoction for alleviating pant were given enema decoction for alleviating pant 8.6 g/kg,3.2 g/kg,and 1.7 g/kg by enema,respectively,all once daily,continuously treated for 7 days.The pathological morphology of lung tissues in the rats were observed by HE staining,the serum levels of tumor necrosis factor-α( TNF-α),interleukin ( IL)-1β,IL-8 and IL-13 were detected by ELISA,the mRNA expressions of AMPK,Unc-51-like autophagy-activated kinase (ULK1),Beclin1 and autophagy protein p62 were detected by RT-PCR,and the protein expressions of phosphorylated AMPK (p-AMPK),rapamycin protein kinase (mTOR),ULK1,Beclin1,microtubule-associated protein light chain 3 ( LC3Ⅱ/LC3Ⅰ),and p62 protein in lung tissues were detected by Western blot.Results The bronchial mucosa and tube wall were infiltrated by a large number of inflammatory cells with compensatory emphysema and pulmonary bulla formation in the model group;the inflammatory cells were reduced,and pulmonary bullas were significantly reduced in the treatment groups,among which the improvements were more significantly in the high dose group of enema decoction for alleviating pant and dexamethasone group.The serum levels of TNF-α,IL-1β,IL-8 and IL-13 in the model group were significantly higher than those in the blank group (all P<0.05);the serum levels of TNF-α,IL-1β,IL-8 and IL-13 in the medium,and high dose groups of enema decoction for alleviating pant and dexamethasone group and the serum levels of IL-1β and IL-13 in the low dose group of enema decoction for alleviating pant were significantly lower than those in the model group (all P<0.05).Compared with the blank group,the relative mRNA expressions of AMPK,ULK1,Beclin1 and the rela-tive protein expressions of p-AMPK,ULK1,Beclin1,LC3Ⅱ and LC3Ⅱ/LC3Ⅰwere significantly decreased ( all P<0.05 ),while the relative mRNA expression of p62 and relative protein expressions of mTOR and p62 were significantly in-creased in the model group (all P<0.05).Compared with the model group,the relative mRNA expressions of AMPK,ULK1 ( except for the medium dose group of enema decoction for alleviating pant),Beclin1 and the relative protein expres-sions of p-AMPK,ULK1,Beclin1,LC3Ⅱ and were significantly increased (all P<0.05),while the relative mRNA ex-pression of p62 and relative protein expressions of mTOR and p62 were significantly decreased in the medium,and high dose groups of enema decoction for alleviating pant and dexamethasone group (all P<0.05).Conclusion Enema decoction for alleviating pant can enhance autophagy and inhibit inflammatory response in lung tissues of COPD rats,and its mecha-nism may be related to the activation of AMPK pathway.
chronic obstructive pulmonary diseaseenema decoction for alleviating pantautophagyAMP-activated protein kinaseinflammation
万方数据
维普
