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中国临床药理学与治疗学 ›› 2025, Vol. 30 ›› Issue (1): 42-50.doi: 10.12092/j.issn.1009-2501.2025.01.005

• 基础研究 • 上一篇    下一篇

大黄素调节SIRT1/AMPK信号通路对肺炎克雷伯菌致重症肺炎大鼠细胞自噬和凋亡的影响

宋晓萍1,刘萍萍2,刘晓琳3,郑艳1,孙滨1,丁健1,朱元祺4,李峻峰5   

  1. 1青岛大学附属青岛市海慈医院(青岛市中医医院)检验科,青岛  266033,山东;2青岛市第六人民医院检验科,青岛  266033,山东;3青岛市疾病预防控制中心,青岛  266033,山东;4青岛大学附属医院检验科,青岛  266003,山东;5青岛市妇女儿童医院,青岛  266034,山东

  • 收稿日期:2024-01-24 修回日期:2024-02-26 出版日期:2025-01-26 发布日期:2025-01-02
  • 通讯作者: 李峻峰,硕士,副主任医师,从事中西医结合治疗小儿呼吸系统疾病研究。 E-mail: lijunfeng008@163.com
  • 作者简介:宋晓萍,硕士,从事细菌耐药机制、分子流行病学、中药抗感染机制研究。 E-mail: songxpi@163.com
  • 基金资助:
    山东省医药卫生科技发展计划项目(202101060054);山东省中医药科技项目(Z-2023071)

Effects of emodin on autophagy and apoptosis in rats with severe pneumonia caused by Klebsiella pneumoniae by regulating SIRT1/AMPK signaling pathway

SONG Xiaoping1, LIU Pingping2, LIU Xiaolin3, ZHENG Yan1, SUN Bin1, DING Jian1, ZHU Yuanqi4, LI Junfeng5   

  1. 1 Department of Clinical Laboratory, Qingdao Haici Hospital (Qingdao Hospital of Traditional Chinese Medicine), Affiliated to Qingdao University, Qingdao 266033, Shandong, China; 2 Department of Clinical Laboratory, Qingdao Sixth People's Hospital, Qingdao 266033, Shandong, China; 3 Qingdao Center for Disease Control and Prevention, Qingdao 266033, Shandong, China; 4 Department of Clinical Laboratory, Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong, China; 5 Qingdao Women and Children's Hospital, Qingdao 266034, Shandong, China
  • Received:2024-01-24 Revised:2024-02-26 Online:2025-01-26 Published:2025-01-02

摘要:

目的:探讨大黄素对肺炎克雷伯菌(K. pneumoniae)致重症肺炎(KP)大鼠细胞自噬和凋亡的影响及可能的机制。方法:通过感染K. pneumoniae建立KP大鼠模型并用大黄素处理。将大鼠分为假手术组、KP组、低浓度大黄素组(20 mg/kg)、中浓度大黄素组(40 mg/kg)、高浓度大黄素组(80 mg/kg)、大黄素+sirtinol[沉默信息调节因子(SIRT1)活性抑制剂]组(80 mg/kg大黄素+30 μL/kg sirtinol);血气分析仪检测动脉血二氧化碳分压(PaCO2)、动脉氧分压(PaO2)、动脉血氧饱和度(SaO2);瑞氏-吉姆萨染色(Wright-Giemsa)测定支气管肺泡灌洗液(BALF)中白细胞和中性粒细胞计数;HE染色检测各组肺组织病理变化;ELISA检测各组肺组织中IL-6、TNF-α、IL-1β表达;电镜扫描观察各组肺组织中细胞自噬情况;免疫荧光染色观察LC3B在肺组织中的表达情况;TUNEL法检测各组大鼠肺组织细胞凋亡变化;Western blot检测肺组织中SIRT1、腺苷单磷酸活化蛋白激酶(AMPK)、LC3-II、LC3-I、c-caspase-3、caspase-3蛋白表达。结果:K. pneumoniae诱导的肺炎大鼠肺组织呈现严重损伤,肺部炎性浸润和炎性细胞因子释放增加,动脉血中PaO2和SaO2水平降低,PaCO2水平升高,BALF中白细胞和中性粒细胞计数升高,细胞凋亡率和c-caspase-3/caspase-3水平增加,细胞自噬以及自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ水平降低(均P<0.05)。大黄素处理后激活SIRT1/AMPK信号通路,升高动脉血中PaO2和SaO2水平,降低PaCO2水平,抑制炎症反应,抑制肺组织细胞凋亡,恢复细胞自噬水平(均P<0.05),而使用SIRT1抑制剂sirtinol抑制SIRT1/AMPK信号通路后部分逆转了大黄素对KP大鼠的治疗作用(P<0.05)。结论:大黄素可能通过激活SIRT1/AMPK通路进而增强大鼠肺组织细胞自噬,抑制大鼠肺组织细胞凋亡,可能为KP提供潜在的治疗选择。

关键词: 大黄素, 沉默信息调节因子/腺苷单磷酸活化蛋白激酶信号通路, 肺炎克雷伯菌, 重症肺炎, 大鼠, 细胞自噬, 凋亡

Abstract:

AIM: To investigate the effects of emodin on autophagy and apoptosis in rats with severe pneumonia (KP) caused by K. pneumoniae and its possible mechanism. METHODS: The KP rat model was established by infecting K pneumonia was treated with Emodin. The rats were grouped into Sham surgery group, KP group, low concentration Emodin group, medium concentration Emodin group, high concentration Emodin group, and Emodin+sirtinol (SIRT1 activity inhibitor) group; Arterial partial pressure of carbon dioxide (PaCO2), arterial partial pressure of oxygen (PaO2) and arterial oxygen saturation (SaO2) were measured by blood gas analyzer; the white blood cells and neutrophils in bronchoalveolar lavage fluid (BALF) were measured by Wright-Giemsa staining; HE staining was applied to detect pathological changes in lung tissue in each group; ELISA was applied to detect the expression of IL-6, TNF-α, and IL-1β in lung tissues of each group; electron microscopy scanning was applied to observe the autophagy of cells in lung tissues of each group; the expression of LC3B in lung tissues was observed by immunofluorescence staining; TUNEL method was applied to detect changes in cell apoptosis in lung tissue of rats in each group; Western blot was applied to detect the expression of silent information regulatory factor (SIRT1), adenosine monophosphate activated protein kinase (AMPK), LC3-II, LC3-I, c-caspase-3, and caspase-3 proteins in lung tissue. RESULTS: K. pneumoniae caused severe lung tissue damage in rats with pneumonia, increased inflammatory infiltration and cytokine release in the lungs, arterial blood PaO2 and SaO2 levels decreased, PaCO2 levels increased, white blood cells and neutrophils count increased in BALF, increased cell apoptosis rate and c-caspase-3/caspase-3 level, and the cell autophagy and the levels of autophagy related proteins LC3-II/LC3-I were decreased (all P<0.05), after Emodin treatment, SIRT1/AMPK signaling pathway was activated, PaO2 and SaO2 levels in arterial blood were increased, PaCO2 levels was decreased, inflammatory reaction was inhibited, cell apoptosis in lung tissue was inhibited (all P<0.05), and cell autophagy level was restored, sirtinol, a SIRT1 inhibitor, partially reversed the therapeutic effect of Emodin on KP rats after inhibiting SIRT1/AMPK signaling pathway (P<0.05). CONCLUSION: Emodin may enhance autophagy of lung tissue cells and inhibit apoptosis of rat lung tissue cells by activating SIRT1/AMPK pathway, which may provide potential therapeutic options for KP.

Key words: emodin, silent information regulator/adenosine monophosphate-activated protein kinase signaling pathway, Klebsiella pneumoniae, severe pneumonia, rats, cell autophagy, apoptosis

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