绿原酸对小鼠急性肺损伤的保护作用研究
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摘要
本文探讨了绿原酸对补体旁路激活致小鼠急性肺损伤的保护作用及可能的作用机制。将32只KM小鼠随机分为正常对照组、模型组、白藜芦醇组、绿原酸组,预防给药7 d后,用特异性补体旁路激活蛋白眼镜蛇毒因子(CVF)尾静脉注射复制小鼠急性肺损伤模型。测定肺含水量、肺组织匀浆中髓过氧化物酶(MPO)活性、支气管肺泡灌洗液(BALF)中细胞数目和蛋白含量,ELISA法检测BALF和血清中白介素-6(IL-6)、肿瘤坏死因子(TNF-α)、P选择素(P-selectin)和细胞间黏附分子-1(ICAM-1)的含量,HE染色法观察肺组织病理形态学变化,免疫组化法测定肺组织中NF-κB p65的磷酸化水平。研究发现,绿原酸可以降低小鼠BALF中蛋白含量、炎性细胞数目、IL-6和TNF-α含量以及肺组织匀浆中MPO活性,并可显著降低血清中IL-6、TNF-α及P-selectin、ICAM-1水平;病理形态学显示绿原酸可以显著抑制炎性细胞浸润,免疫组化结果显示绿原酸可显著抑制小鼠NF-κB p65的磷酸化水平。以上结果说明绿原酸可明显减轻补体旁路激活诱导的小鼠急性肺损伤,其机制可能与抑制NF-κB p65的磷酸化及降低炎症反应程度有关。
This study aimed to explore the protective effect and mechanism of chlorogenic acid( CGA) on acute lung injury( ALI) in mice induced by activation of complement alternative pathway. 32 healthy Kunming mice were randomly divided into four groups: the normal group,the model group,the resveratrol group and the CGA group,which received 7-day intragastric administration respectively. Then Cobra venom factor( CVF) was used to activate complement alternative pathway of Kunming mice by intravenous injection. Lung water content,BALF cell count,the protein content in bronchoalveolar lavage fluid( BALF) and myeloperoxidase( MPO) activity of lung homogenate were measured. The concentration of IL-6,TNF-α,P-selectin and ICAM-1 in BALF and serum was determined by ELISA. The pathological change of lung tissue was observed by HE staining. The phosphorylation of NF-κB p65 in lung tissues was checked by immunohistochemistry. The results showed that CGA reduced the protein concentration,the inflammatory cell count,the content of IL-6 and TNF-α in BALF and MPO activity of lung homogenate obviously. CGA also decreased the level of IL-6,TNF-α,P-selectin and ICAM-1 in serum,reduced the pulmonary inflammatory cell infiltration,and inhibited the phosphorylation of NF-κB p65 in ALI mice induced by activation of complement alternative pathway significantly. With these results,it is concluded that CGA can alleviate inflammatory response of ALI mice induced by activation of complement alternative pathway. The mechanism is highly related to the down-regulation of phosphorylation of NF-κB p65 and the decrease of inflammatory response.
This study aimed to explore the protective effect and mechanism of chlorogenic acid( CGA) on acute lung injury( ALI) in mice induced by activation of complement alternative pathway. 32 healthy Kunming mice were randomly divided into four groups: the normal group,the model group,the resveratrol group and the CGA group,which received 7-day intragastric administration respectively. Then Cobra venom factor( CVF) was used to activate complement alternative pathway of Kunming mice by intravenous injection. Lung water content,BALF cell count,the protein content in bronchoalveolar lavage fluid( BALF) and myeloperoxidase( MPO) activity of lung homogenate were measured. The concentration of IL-6,TNF-α,P-selectin and ICAM-1 in BALF and serum was determined by ELISA. The pathological change of lung tissue was observed by HE staining. The phosphorylation of NF-κB p65 in lung tissues was checked by immunohistochemistry. The results showed that CGA reduced the protein concentration,the inflammatory cell count,the content of IL-6 and TNF-α in BALF and MPO activity of lung homogenate obviously. CGA also decreased the level of IL-6,TNF-α,P-selectin and ICAM-1 in serum,reduced the pulmonary inflammatory cell infiltration,and inhibited the phosphorylation of NF-κB p65 in ALI mice induced by activation of complement alternative pathway significantly. With these results,it is concluded that CGA can alleviate inflammatory response of ALI mice induced by activation of complement alternative pathway. The mechanism is highly related to the down-regulation of phosphorylation of NF-κB p65 and the decrease of inflammatory response.
引文
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14 Lin ZS,Ku CF,Guan YF,et al.Dihydro-resveratrol ameliorates lung injury in rats with cerulein-induced acute pancreatitis[J].Phytother Res,2016,30:663-670.
15 Torun AC,Tutuncu S,Ustun B,et al.A study of the therapeutic effects of resveratrol on blunt chest rauma-induced acute lung injury in rats and the potential role of endocan as a biomarker of inflammation[J].Inflammation,2017,40:1803-1810.
16 Li CS(李朝胜),Sun QY(孙黔云).Effect of three chemical molecules on adhesion molecules expression in HEMCs induced by activated complement alternative pathway[J].Chin Pharmacol Bull(中国药理学通报),2015,31:1421-1426.
2 Sun QY(孙黔云).Application of cobra venom factor in life sciences[J].Chin Bull Life Sci(生命科学),2016,28:22-26.
3 Wu WH(吴卫华),Kang Z(康桢),Ouyang DS(欧阳冬生),et al.Progresses in the pharmacology of chlorogenic acid[J].Nat Prod Res Dev(天然产物研究与开发),2006,18:691-694.
4 Zhou Y(周英),Li M(李敏),Sun QY(孙黔云).Effect of chlorogenic acid,caffeic acid,and ferulic acid oninhibition of inflammatory response of HMECs induced by activated complement alternative pathway[J].Chin Pharmacol Bull(中国药理学通报),2016,32:1723-1728.
5 Sun QY,Chen G,Guo H,et al.Prolonged cardiac xenograft survival in guinea pig-to-rat model by a highly active cobra venom factor[J].Toxicon,2003,42:257-262.
6 Guo J(郭静),Li M(李敏),Yang FM(杨付梅),et al.Inflammatory mechanism of acute lung injury in mice induced by activation of complement alternative pathway[J].Chin Pharmacol Bull(中国药理学通报),2016,32:1521-1526.
7 Chen XJ(陈向军),Ma LJ(马李杰),Hou SJ(侯绍杰),et al.Protective effect of resveratrol on acute lung injury induce by lipopolysaccharide[J].Chin J Lung Dis:Electronic Edition(中华肺部疾病杂志:电子版),2013,6:20-24.
8 Li HX(李洪辛),Jiang HB(姜洪彬),Liu SN(刘姝男),et al.Effect and mechanism of chlorogenic acid in allergic rhinitis mice[J].Chin J Biochem Pharm(中国生化药物杂志),2015,35:1-5.
9 Ou YY,Jiang Y,Li H,et al.Polysaccharides from arnebia euchroma ameliorated endotoxic fever and acute lung injury in rats through inhibiting complement system[J].Inflammation,2017,40:275-284.
10 Sun S,Jiang Y,Wang R,et al.Treatment of paraquat-induced lung injury with an anti-C5a antibody:potential clinical application[J].Crit Care Med,2018.
11 Li RQ(李瑞奇).Study on intervening effect of chlorogenic acid on cerebral ischemia reperfusion rats and mice model[D].Zhengzhou:Henan college of Chinese medicine(河南中医学院),2014.
12 Song Z(宋卓),Huang K(黄康),Huang L(黄林),et al.Effect of chlorogenic acid at high dose on expression of hepatic inflammatory cytokines mRNA induced by lipopolysaccharides[J].Acta Nutri Sin(营养学报),2014,36:481-485.
13 Jiang L,Zhang L,Kang K,et al.Resveratrol ameliorates LPSinduced acute lung injury via NLRP3 inflammasome modulation[J].Biomed Pharmacother,2016,84:130-138.
14 Lin ZS,Ku CF,Guan YF,et al.Dihydro-resveratrol ameliorates lung injury in rats with cerulein-induced acute pancreatitis[J].Phytother Res,2016,30:663-670.
15 Torun AC,Tutuncu S,Ustun B,et al.A study of the therapeutic effects of resveratrol on blunt chest rauma-induced acute lung injury in rats and the potential role of endocan as a biomarker of inflammation[J].Inflammation,2017,40:1803-1810.
16 Li CS(李朝胜),Sun QY(孙黔云).Effect of three chemical molecules on adhesion molecules expression in HEMCs induced by activated complement alternative pathway[J].Chin Pharmacol Bull(中国药理学通报),2015,31:1421-1426.