人参皂苷Rg1通过调控AMPK/NLRP3通路介导的细胞焦亡抑制大鼠肺纤维化研究  

Ginsenoside Rg1 inhibits pulmonary fibrosis in rats by regulating AMPK/NLRP3 pathway-mediated pyroptosis

作　　者：邓宏哲 陈昆[1] 李鹏 朱清海 DENG Hong-zhe;CHEN Kun;LI Peng;ZHU Qing-hai(Department of General Laparoscopy,Weight Loss and Metabolic Surgery,Zhumadian Central Hospital Directly Affiliated to Huanghuai College,Zhumadian 463000,China;School of Chemistry and Pharmaceutical Engineering,Huanghuai College,Zhumadian 463003,China)

机构地区：[1]黄淮学院直属附属驻马店市中心医院普外腹腔镜、减重与代谢外科,河南驻马店463000 [2]黄淮学院化学与制药工程学院,河南驻马店463003

出　　处：《中草药》2023年第3期841-848,共8页Chinese Traditional and Herbal Drugs

基　　金：河南省高等学校自然科学重点科研项目(21B320122)。

摘　　要：目的探究人参皂苷Rg_(1)对大鼠肺纤维化(pulmonary fibrosis,PF)的影响及作用机制。方法50只雄性SD大鼠随机分为对照组、模型组、人参皂苷Rg_(1)(72 mg/kg)组、腺苷酸活化蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)激动剂(200 mg/kg)组、人参皂苷Rg_(1)(72 mg/kg)+AMPK抑制剂(20 mg/kg)组,每组10只。除对照组外,其余各组大鼠气管内注射博来霉素(5 mg/kg)构建大鼠PF模型。造模成功后2 d开始给药,连续给药28 d后检测大鼠肺功能指标;采用苏木素-伊红(HE)、Masson染色观察肺组织病理变化;采用免疫组化检测肺组织I型胶原(collagen I)和α-肌动球蛋白(α-smooth muscle actin,α-SMA)表达;采用试剂盒测定肺组织羟脯氨酸(hydroxyproline,Hyp)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、IL-1β和IL-18水平;采用Western blotting检测肺组织p-AMPK/AMPK、NOD样蛋白3(NOD-like receptor protein 3,NLRP3)、半胱氨酸天冬氨酸蛋白酶-1(cysteinasparate protease-1,Caspase-1)、消皮素D(gasdermin D,GSDMD)、IL-1β和IL-18蛋白表达。结果与模型组比较,人参皂苷Rg_(1)和AMPK激动剂组大鼠肺功能指标明显升高(P<0.05),肺纤维化程度改善,肺指数及肺组织Hyp、TNF-α、IL-6、IL-1β和IL-18水平均明显降低(P<0.05),肺组织collagen I、α-SMA、NLRP3、cleaved Caspase-1/pro Caspase-1、GSDMD-N/GSDMD、IL-1β和IL-18蛋白表达水平均显著降低(P<0.05),p-AMPK/AMPK蛋白表达升高(P<0.05)。与人参皂苷Rg_(1)组比较,人参皂苷Rg_(1)+AMPK抑制剂组大鼠肺功能指标降低(P<0.05),肺纤维化程度加重,肺指数及肺组织Hyp、TNF-α、IL-6、IL-1β和IL-18水平均明显增加(P<0.05),肺组织collagen I、α-SMA、NLRP3、cleaved Caspase-1/pro Caspase-1、GSDMD-N/GSDMD、IL-1β和IL-18蛋白表达升高(P<0.05),p-AMPK/AMPK表达降低(P<0.05)。结论人参皂苷Rg_(1)能够抑制AMPK/NLRP3介导的细胞焦亡改善博来霉素诱导的大鼠PF。Objective To investigate the effect and mechanism of ginsenoside Rg_(1)on pulmonary fibrosis(PF)in rats.Methods A total of 50 male SD rats were randomly divided into control group,model group,ginsenoside Rg_(1)(72 mg/kg)group,adenosine monophosphate activated protein kinase(AMPK)agonist(200 mg/kg)group and ginsenoside Rg_(1)(72 mg/kg)+AMPK inhibitor group(20 mg/kg),with 10 rats in each group.Except for the control group,rats in the other groups were injected with bleomycin(5 mg/kg)into trachea to establish a rat model of PF.The drug was administered two days after successful modeling.After continuous administration for 28 d,pulmonary function indexes of rats were detected;HE and Masson staining were used to observe pathological changes of lung tissue;Immunohistochemistry was used to detect the expressions of collagen I andα-smooth muscle actin(α-SMA)in lung tissue;Levels of hydroxyproline(Hyp),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1βand IL-18 in lung tissue were measured by kit.p-AMPK/AMPK,NOD-like receptor protein 3(NLRP3),cysteine-aspartate protease-1(Caspase-1),gasdermin D(GSDMD),IL-1βand IL-18 protein expressions in lung tissue were detected by Western blotting.Results Compared with model group,lung function indexes of rats in ginsenoside Rg_(1)group and AMPK agonist group were significantly increased(P<0.05),degree of pulmonary fibrosis was improved,lung index and levels of Hyp,TNF-α,IL-6,IL-1βand IL-18 in lung tissue were significantly decreased(P<0.05),collagen I,α-SMA,NLRP3,cleaved Caspase-1/pro Caspase-1,GSDMD-N/GSDMD,IL-1βand IL-18 protein expressions in lung tissues were significantly decreased(P<0.05),while p-AMPK/AMPK protein expression was increased(P<0.05).Compared with ginsenoside Rg_(1)group,lung function index in ginsenoside Rg_(1)+AMPK inhibitor group was decreased(P<0.05),pulmonary fibrosis was aggravated,lung index and levels of Hyp,TNF-α,IL-6,IL-1βand IL-18 in lung tissue were significantly increased(P<0.05),collagen I,α-SMA,NLRP3,cleansed caspase-1/pro caspase-1

关 键 词：人参皂苷Rg_(1) 肺纤维化 博来霉素 细胞焦亡 AMPK/NLRP3通路 

分 类 号：R285.5[医药卫生—中药学]