[1]樊秋月,姜永红,陈一柳.清肺通络方通过胱天蛋白酶-1/Gasdermin D蛋白细胞焦亡途径对支原体肺炎小鼠的干预机制[J].陕西中医,2023,(4):416-420.[doi:DOI:10.3969/j.issn.1000-7369.2023.04.002]
　FAN Qiuyue,JIANG Yonghong,CHEN Yiliu.Intervention effects of Qingfei Tongluo recipe on Caspase-1/GSDMD cell death pathway in mice with mycoplasma pneumonia[J].,2023,(4):416-420.[doi:DOI:10.3969/j.issn.1000-7369.2023.04.002]
点击复制

清肺通络方通过胱天蛋白酶-1/Gasdermin D蛋白细胞焦亡途径对支原体肺炎小鼠的干预机制
分享到：

《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2023年4期
页码:
416-420
栏目:
基础研究
出版日期:
2023-04-05

文章信息/Info

Title:
Intervention effects of Qingfei Tongluo recipe on Caspase-1/GSDMD cell death pathway in mice with mycoplasma pneumonia
作者:
樊秋月; 姜永红; 陈一柳
(上海中医药大学附属龙华医院儿科,上海 200032)
Author(s):
FAN Qiuyue; JIANG Yonghong; CHEN Yiliu
(Pediatric Department,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)
关键词:
支原体肺炎;  清肺通络方;  细胞焦亡;  胱天蛋白酶-1;  Gasdermin D蛋白;  小鼠
Keywords:
Mycoplasma pneumonia;  Qingfei Tongluo recipe;  Pyroptosis;  Caspase-1;  GSDMD;  Mice
分类号:
R 563.1
DOI:
DOI:10.3969/j.issn.1000-7369.2023.04.002
文献标志码:
A
摘要:
目的:探讨清肺通络方通过胱天蛋白酶-1(Caspase-1)/Gasdermin D(GSDMD)蛋白细胞焦亡途径对支原体肺炎小鼠干预机制。方法:将30只BALB/c小鼠随机分为空白对照组、模型组、清肺通络方组,每组10只。采用肺炎支原体菌株滴鼻法建立支原体肺炎小鼠模型。造模同时予灌胃治疗,连续治疗3 d,实验第4天处死各组小鼠。PCR检测小鼠肺组织MP-DNA含量,HE染色观察小鼠肺组织病理改变,Western blot检测胱天蛋白酶-1前体(pro-Caspase-1)、Caspase-1、GSDMD、Gasdermin D氮端活性结构域多肽(GSDMD-N)表达,ELISA检测肺泡灌洗液中白介素-1β(IL-1β)、白介素-18(IL-18)含量,生化法检测肺组织细胞乳酸脱氢酶(LDH)和Na+-K+-ATP酶活性。结果:镜下观察模型组小鼠肺间质出现炎性细胞浸润、渗出明显。与空白对照组比较,模型组小鼠pro-Caspase-1、Caspase-1、GSDMD、GSDMD-N表达及IL-1β、IL-18含量均升高,LDH活性上升,Na+-K+-ATP酶活性下降(均P<0.05); 与模型组比较,清肺通络方组小鼠肺间质炎症细胞浸润明显减轻,肺组织中Caspase-1、GSDMD、GSDMD-N表达及IL-1β、IL-18含量均降低(均P<0.05),LDH活性下降,Na+-K+-ATP酶活性上升(均P<0.05); 两组pro-Caspase-1表达比较差异无统计学意义(P>0.05)。结论:清肺通络方通过抑制Caspase-1/GSDMD通路,缓解细胞焦亡,减轻小鼠肺组织炎症反应。
Abstract:
Objective:To investigate the intervention effect of Qingfei Tongluo recipe on mycoplasma pneumonia(MP)mice through Caspase-1/GSDMD cell pyrodeath pathway.Methods:30 BALB/c mice were randomly assigned to an blank control group,model group and Qingfei Tongluo recipe group,with 10 mice in each group.The mouse model of mycoplasma pneumoniae was established by nasal drip of mycoplasma pneumoniae strains.The mice in each group were killed on the fourth day of the experiment.The content of MP-DNA in mouse lung tissue was detected by PCR.The pathological changes of lung tissue in each group were observed by HE staining.The expression of pro-Caspase-1,Caspase-1,GSDMD and GSDMD-N was detected by Western blot.The content of IL-1β and IL-18 in alveolar lavage fluid was detected by ELISA.The activity of LDH and Na+-K+-ATPase in lung tissue cells were detected by biochemical method.Results:The infiltration and exudation of inflammatory cells in lung interstitium of model group were observed under microscope.Compared with blank control group,the expressions of pro-Caspase-1,Caspase-1,GSDMD,GSDMD-N and the contents of IL-1β and IL-18 were increased in model group,while the activity of LDH was increased and the activity of Na+-K+-ATPase was decreased(P<0.05).Compared with model group,the infiltration of lung interstitial inflammatory cells in Qingfei Tonglu recipe group was significantly reduced,the expressions of Caspase-1,GSDMD,GSDMD-N and the contents of IL-1β and IL-18 in lung tissue were decreased(P<0.05),the activity of LDH was decreased,and the activity of Na+-K+-ATPase was increased(P<0.05).There was no significant difference in pro-Caspase-1 expression between the two groups(P>0.05).Conclusion:Qingfei Tongluo recipe alleviates pyrodeath and lung tissue inflammation in mice by inhibiting Caspase-1/GSDMD pathway.

参考文献/References:

[1] Waites KB,Crabb DM,Bing X,et al.In vitro susceptibilities to and bactericidal activities of garenoxacin(BMS-284756)and other antimicrobial agents against human mycoplasmas and ureaplasmas[J].Antimicrob Agents Chemother,2003,47(1):161-165.
[2] 毕潇文,李 静.支气管哮喘合并肺炎支原体感染患儿血清白介素-6、25羟基维生素D、肿瘤坏死因子水平变化临床研究[J].陕西医学杂志,2022,51(1):63-66.
[3] 蔡伟娜,周隆辉,侯文静.小儿支原体肺炎并发支气管哮喘应用头孢曲松联合甲强龙冲击疗法的临床分析及对患儿免疫因子影响[J].新疆医学,2021,51(12):1371-1374.
[4] Beeton ML,Zhang X,Uldum SA,et al.Mycoplasma pneumoniae infections,11 countries in Europe and Israel,2011 to 2016[J].Euro Surveill,2020,25(2):13-39.
[5] 赵晓阳,姜永红,肖 臻,等.清肺通络方调控NLRP3炎症小体对支原体肺炎影响的细胞实验研究[J].辽宁中医杂志,2021,48(12):176-183.
[6] 张静雯,王 利,赵 敏,等.核苷酸结合寡聚化结构域样受体蛋白3介导细胞焦亡在痛风性关节炎中的作用机制研究[J].陕西医学杂志,2022,51(5):561-565.
[7] 唐颖慧,叶苗青,何瑾瑜,等.化脂复肝颗粒通过抑制TLR4/NF-κB/NLRP3减轻Kupffer细胞炎症改善非酒精性脂肪性肝病机制研究[J].陕西中医,2022,43(10):1359-1363.
[8] 姜永红,虞坚尔,姜之炎.从“肺络”探讨小儿支原体肺炎的防治[J].天津中医药,2012,29(1):52-53.
[9] 姜永红,虞坚尔,姜之炎.从络病理论解析小儿支原体肺炎及其变证[J].上海中医药杂志,2013,47(5):27-28.
[10] 宋 瑶,姜永红,姜之炎,等.清肺通络方对肺炎支原体肺炎小鼠体内氧化应激的影响[J].现代中西医结合杂志,2021,30(11):1141-1146.
[11] 谭 丹,姜之炎.中医药治疗儿童肺炎支原体肺炎的机制研究进展[J].中华中医药学刊,2018,36(6):1403-1406.
[12] 周其其,闵 清,白育庭.细胞焦亡与肺部疾病的研究进展[J].湖北科技学院学报:医学版,2021,35(4):364-368.
[13] 丁华胜,黄 燕,王凤媛.丹皮酚对心肌缺血再灌注损伤保护中Toll样受体4、心肌焦亡影响的作用机制[J].陕西中医,2021,42(12):1659-1662.
[14] Liu F,Liu T,Sun M,et al.Maxing Shigan decoction mitigates mycoplasma pneumonia-induced pyroptosis in A549 cells via the NLRP3 inflammasome[J].Infect Drug Resist,2021,14:859-867.
[15] 赵 卓,刘 林,宋 囡,等.半夏泻心汤对溃疡性结肠炎大鼠NLRP3/Caspase-1细胞焦亡信号通路的影响[J].中国实验方剂学杂志,2022,28(16):29-34.
[16] Yu P,Zhang X,Liu N,et al.Pyroptosis:Mechanisms and diseases[J].Signal Transduct Target Ther,2021,6(1):128.
[17] 任秋月,姚蓉飞,杨荣禄,等.抵挡汤对高糖环境下泡沫细胞焦亡炎症级联反应的作用机制[J].中国实验方剂学杂志,2022,28(11):8-15.
[18] Jorgensen I,Zhang Y,Krantz BA,et al.Pyroptosis triggers pore-induced intracellular traps(PITs)that capture bacteria and lead to their clearance by efferocytosis[J].Journal of Experimental Medicine,2016,213(10):2113-2128.
[19] Ding J,Wang K,Liu W,et al.Pore-forming activity and structural autoinhibition of the gasdermin family[J].Nature,2016,535(7610):111-116.
[20] Coll RC,Robertson AA,Chae JJ,et al.A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases[J].Nat Med,2015,21(3):248-255.
[21] He W,Wan H,Hu L,et al.GasderminD is an executor of pyroptosis and required for interleukin-1β secretion[J].Cell Research,2015,25(12):1285-1298.
[22] 杨 迪,郭 越,张 鹏,等.阿司匹林调控NLRP3/Caspase-1/GSDMD通路介导的焦亡途径减轻脂多糖诱导的急性肺损伤研究[J].中国药学杂志,2023,58(4):347-355.
[23] 王雪梅,梁志庆,王蓓蓓,等.麻杏止嗽散对咳嗽变异型哮喘模型小鼠IFN-γ、IL-8水平的影响[J].云南中医学院学报,2022,45(6):82-86.
[24] 任韩雯婧,张 川,乐 涛,等.ICU重症肺部感染患者血清IL-6、IL-1β、IL-8和TNF-α水平变化及其与预后的关系[J].热带医学杂志,2022,22(10):1371-1375,1390,1461.
[25] 欧小琴.203例儿童肺炎支原体肺炎相关因素分析[J].四川医学,2021,42(10):1017-1021.
[26] 姜永红,陈一柳,樊秋月,等.清肺通络方调控Caspase-1对肺炎支原体诱导人正常肺上皮细胞BEAS-2B焦亡的干预作用[J].广州中医药大学学报,2023,40(2):450-460.
[27] Kryvenko V,Vagin O,Dada LA,et al.Maturation of the Na+-K+-ATPase in the endoplasmic reticulum in health and disease[J].The Journal of Membrane Biology,2021,254(5):447-457.
[28] 翟玉荣,波 刘,莫镇涛,等.苦瓜提取物对糖尿病大鼠肾皮质Na+-K+-ATP 酶活性的影响[J].遵义医学院学报,2015,38(2):121-124.

相似文献/References:

[1]韩 鹏.小儿豉翘清热颗粒联合阿奇霉素对支原体肺炎患儿机体 炎症反应及免疫功能影响研究[J].陕西中医,2019,(4):424.
[2]张 锦,黄 晶.桑白皮汤加减治疗痰热闭肺型小儿支原体肺炎临床研究[J].陕西中医,2019,(6):705.
[3]马文明,耿 伟,吴红胜.升麻汤联合阿奇霉素对支原体肺炎患儿炎症指标及免疫功能的影响*[J].陕西中医,2020,(10):1379.[doi:DOI:10.3969/j.issn.1000-7369.2020.10.010]

备注/Memo

备注/Memo:
基金项目:上海市自然科学基金资助项目(20ZR1458800); 上海市进一步加快中医药传承创新发展三年行动计划项目[ZY(2021-2023)-0206-01]; 上海市十三五临床重点专科建设项目(SHSLCZDZK04102)
更新日期/Last Update: 2023-04-10