{"title":"Andrographolide Derivative AL-1 Ameliorates LPS-induced Acute Lung Injury by Inhibiting NLRP3 Inflammasome and Lung Permeability.","authors":"Tangjia Li, Chu Zhang, Yuke Wei, Haijing Zhong, Luchen Shan, Pei Yu, Yuqiang Wang, Lipeng Xu","doi":"10.2174/1381612828666220729094806","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Acute lung injury (ALI) is a serious respiratory disease with a high mortality rate, and there is an urgent need for a more effective treatment strategy. Andrographolide derivative AL-1 has been identified to possess anti-inflammatory activity. However, whether it could reduce LPS-induced lung injury in mice through inhibiting NLRP3 inflammasome activation and protecting lung permeability has not yet been elucidated. In the present research, we investigated the protective effect of AL-1 on ALI mice and demonstrated the potential mechanisms.</p><p><strong>Methods: </strong>Male Balb/c mice were anesthetized with isoflurane, and ALI mice were induced by intratracheal instillation of LPS. The mice were euthanized after LPS administration for 12 h, then bronchoalveolar lavage fluid (BALF) and lung tissues were collected. The levels of inflammatory factors were measured by ELISA assay, and HE staining and lung injury scoring were used to evaluate the pathological changes in the pulmonary tissues. Immunohistochemistry and immunofluorescence examination were conducted to detect the expression levels of related proteins. Western blot was performed to measure the levels of NLRP3 inflammasome and tight junction proteins.</p><p><strong>Results: </strong>The study indicated that AL-1 effectively alleviated lung injury by reduction of proinflammatory cytokine levels, MPO activity, lung W/D ratio, and total protein levels. Furthermore, AL-1 improved pathological changes in lung tissue and significantly reduced the infiltration of inflammatory cells. Administration with AL-1 markedly inhibited the expression of NLRP3, ASC, Caspase-1, IL-1β, gasdermin D (GSDMD), and VCAM-1 but increased the expression of ZO-1, Occludin, JAM-A, and Claudin-1.</p><p><strong>Conclusion: </strong>Taken together, these results demonstrated that AL-1 ameliorated pulmonary damage by inhibiting the activation of the NLRP3 inflammasome pathway and restoring TJ protein expression.</p>","PeriodicalId":10845,"journal":{"name":"Current pharmaceutical design","volume":null,"pages":null},"PeriodicalIF":2.6000,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current pharmaceutical design","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.2174/1381612828666220729094806","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PHARMACOLOGY & PHARMACY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Acute lung injury (ALI) is a serious respiratory disease with a high mortality rate, and there is an urgent need for a more effective treatment strategy. Andrographolide derivative AL-1 has been identified to possess anti-inflammatory activity. However, whether it could reduce LPS-induced lung injury in mice through inhibiting NLRP3 inflammasome activation and protecting lung permeability has not yet been elucidated. In the present research, we investigated the protective effect of AL-1 on ALI mice and demonstrated the potential mechanisms.
Methods: Male Balb/c mice were anesthetized with isoflurane, and ALI mice were induced by intratracheal instillation of LPS. The mice were euthanized after LPS administration for 12 h, then bronchoalveolar lavage fluid (BALF) and lung tissues were collected. The levels of inflammatory factors were measured by ELISA assay, and HE staining and lung injury scoring were used to evaluate the pathological changes in the pulmonary tissues. Immunohistochemistry and immunofluorescence examination were conducted to detect the expression levels of related proteins. Western blot was performed to measure the levels of NLRP3 inflammasome and tight junction proteins.
Results: The study indicated that AL-1 effectively alleviated lung injury by reduction of proinflammatory cytokine levels, MPO activity, lung W/D ratio, and total protein levels. Furthermore, AL-1 improved pathological changes in lung tissue and significantly reduced the infiltration of inflammatory cells. Administration with AL-1 markedly inhibited the expression of NLRP3, ASC, Caspase-1, IL-1β, gasdermin D (GSDMD), and VCAM-1 but increased the expression of ZO-1, Occludin, JAM-A, and Claudin-1.
Conclusion: Taken together, these results demonstrated that AL-1 ameliorated pulmonary damage by inhibiting the activation of the NLRP3 inflammasome pathway and restoring TJ protein expression.
背景:急性肺损伤(ALI)是一种死亡率高的严重呼吸系统疾病,迫切需要一种更有效的治疗策略。穿心莲内酯衍生物AL-1具有抗炎活性。然而,是否能通过抑制NLRP3炎性体活化、保护肺通透性来减轻lps诱导的小鼠肺损伤尚不清楚。在本研究中,我们研究了AL-1对ALI小鼠的保护作用,并论证了其可能的机制。方法:采用异氟醚麻醉Balb/c雄性小鼠,气管内注入LPS诱导ALI小鼠。LPS给药12 h后处死小鼠,收集支气管肺泡灌洗液(BALF)和肺组织。采用ELISA法检测炎症因子水平,采用HE染色及肺损伤评分法评价肺组织病理变化。免疫组织化学和免疫荧光检测相关蛋白的表达水平。Western blot检测NLRP3炎性体和紧密连接蛋白水平。结果:AL-1通过降低促炎细胞因子水平、MPO活性、肺W/D比和总蛋白水平,有效减轻肺损伤。此外,AL-1改善了肺组织的病理改变,明显减少了炎症细胞的浸润。AL-1显著抑制NLRP3、ASC、Caspase-1、IL-1β、gasdermin D (GSDMD)和VCAM-1的表达,增加ZO-1、Occludin、JAM-A和Claudin-1的表达。结论:综上所述,这些结果表明AL-1通过抑制NLRP3炎症小体通路的激活和恢复TJ蛋白的表达来改善肺损伤。
期刊介绍:
Current Pharmaceutical Design publishes timely in-depth reviews and research articles from leading pharmaceutical researchers in the field, covering all aspects of current research in rational drug design. Each issue is devoted to a single major therapeutic area guest edited by an acknowledged authority in the field.
Each thematic issue of Current Pharmaceutical Design covers all subject areas of major importance to modern drug design including: medicinal chemistry, pharmacology, drug targets and disease mechanism.