[1]郑爱芳,马 涛,李菲菲,等.BPIFB4对香烟烟雾提取物诱导BEAS-2B细胞中NLRP3介导的细胞焦亡影响的实验研究[J].现代检验医学杂志,2026,41(02):63-69.[doi:10.3969/j.issn.1671-7414.2026.02.011]
 ZHENG Aifang,MA Tao,LI Feifei,et al.Experimental Study on the Effect of BPIFB4 on NLRP3 Mediated Pyroptosis in BEAS-2B Cells Induced by Cigarette Smoke Extract[J].Journal of Modern Laboratory Medicine,2026,41(02):63-69.[doi:10.3969/j.issn.1671-7414.2026.02.011]
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BPIFB4对香烟烟雾提取物诱导BEAS-2B细胞中NLRP3介导的细胞焦亡影响的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
63-69
栏目:
论著
出版日期:
2026-03-15

文章信息/Info

Title:
Experimental Study on the Effect of BPIFB4 on NLRP3 Mediated Pyroptosis in BEAS-2B Cells Induced by Cigarette Smoke Extract
文章编号:
1671-7414(2026)02-063-07
作者:
郑爱芳马 涛李菲菲阿衣古杂力·阿布都喀迪尔曹冬冬刘仲山李 黎
喀什地区第一人民医院呼吸与危重症医学科,新疆喀什 844000
Author(s):
ZHENG AifangMA TaoLI FeifeiAYIGUZALI·AbudukadierCAO DongdongLIU ZhongshanLI Li
Department of Respiratory and Critical Care Medicine, the First People’s Hospital of Kashi, Xinjiang Kashi 844000, China
关键词:
慢性阻塞性肺疾病BPI折叠家族B成员4香烟烟雾提取物NOD样受体热蛋白结构域相关蛋白3细胞焦亡
分类号:
R563;R446.8
DOI:
10.3969/j.issn.1671-7414.2026.02.011
文献标志码:
A
摘要:
目的?探究BPI折叠家族B成员4(BPIFB4)对香烟烟雾提取物(CSE)诱导的BEAS-2B细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)介导的细胞焦亡的影响。方法BEAS-2B细胞使用1%、2%、3%浓度的CSE溶液处理24、48、72h,用于筛选CSE最佳诱导模型。BEAS-2B细胞进行慢病毒感染及NLRP3激动剂尼日利亚菌素钠盐处理,细胞分为对照组、CSE组、CSE+过表达BPIFB4组(CSE+BPIFB4)、CSE+过表达对照组(CSE+NC),CSE+BPIFB4+尼日利亚菌素钠盐组和CSE+NC+尼日利亚菌素钠盐组。细胞计数试剂盒8(CCK-8)法检测细胞活力,透射电镜观察细胞超微结构,酶联免疫吸附试验(ELISA)法检测细胞培养上清液中炎症因子水平,实时荧光定量PCR(qRT-PCR)、蛋白印迹法(Westernblotting)检测细胞BPIFB4及NLRP3相关蛋白mRNA和蛋白表达。结果选择3%CSE处理48h作为CSE最佳诱导模型。CSE处理后,与对照组比较,CSE组细胞活力、BPIFB4表达水平降低,细胞膜孔洞增多,细胞培养上清液白细胞介素(IL)-1β、IL-18水平升高(t=5.232~17.234),NLRP3、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸酶1(Caspase-1)、IL-1β、IL-18mRNA和蛋白表达、GasderminD(GSD-MD)mRNA和GSDMD-N蛋白表达水平升高(tmRNA=14.498~20.427,t蛋白=6.245~16.593),差异具有统计学意义(均P<0.05)。与CSE+NC组比较,过表达BPIFB4后CSE+BPIFB4组细胞活力、BPIFB4表达水平升高,细胞膜孔洞减少,细胞培养上清液IL-1β、IL-18水平降低(t=3.827~10.919),NLRP3、ASC、Caspase-1、IL-1β、IL-18mRNA和蛋白表达及GSDMDmRNA和GSDMD-N蛋白表达水平降低(tmRNA=8.102~10.362,t蛋白=8.647~10.947),差异具有统计学意义(均P<0.05)。与CSE+BPIFB4组比较,尼日利亚菌素钠盐处理后CSE+BPIFB4+尼日利亚菌素钠盐组细胞活力、BPIFB4表达水平降低,细胞膜孔洞增多,细胞培养上清液IL-1β、IL-18水平升高(t=3.308~8.930),NLRP3、ASC、Caspase-1、IL-1β、IL-18mRNA和蛋白表达、GSD-MDmRNA和GSDMD-N蛋白表达水平升高(tmRNA=5.759~9.413,t蛋白=3.843~15.702),差异具有统计学意义(均P<0.05)。结论BPIFB4可通过抑制NLRP3表达改善CSE诱导的BEAS-2B细胞焦亡。
Abstract:
Objective To explore the effect of BPI fold containing family B member 4 (BPIFB4) on cigarette smoke extract (CSE)-induced BPIFB4 pyroptosis mediated by NOD like receptor thermal protein domain associated protein 3 (NLRP3) in BE-AS-2B cells. Methods BEAS-2B cells were treated with 1%, 2%, 3% (v/v) CSE solutions for 24, 48 and 72h to identify the opti-mal CSE induction model. BEAS-2B cells were infected with lentivirus and treated with NLRP3 agonist Nigericin sodium salt. Cells were divided into control group, CSE group, CSE+BPIFB4 overexpression group (CSE+BPIFB4), CSE+control overex-pression group (CSE+NC), CSE+BPIFB4+Nigericin sodium salt group and CSE+NC+Nigericin sodium salt group. Cell counting kit 8 (CCK-8) method was used to detect cell viability. Transmission electron microscopy was used to observe cellular ultrastruc-ture. Enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory cytokine levels in cell culture supernatant. Real-time quantitative PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein levels of BPIFB4 and NLRP3 related molecules in cells. Results 3% CSE treatment for 48h was selected as the optimal CSE induction model. Com-pared with the control group, CSE group exhibited reduced cell viability and expression of BPIFB4, while increased cell membrane pores, interleukin (IL)-1β and IL-18 levels in cell culture supernatant(t=5.232~17.234), and the mRNA and pro-tein expression levels of NLRP3, apoptosis associated speck like protein containing CARD (ASC), Cystein containing aspartat specific protease 1 (Caspase-1), IL-1β, IL-18 mRNA and protein and Gasdermin D (GSDMD) mRNA and GSDMD-N protein (tmRNA=14.498~20.427,tprotein=6.245~16.593), the difference were statistically significant (all P<0.05). Compared with CSE+NC group, CSE+BPIFB4 group exhibited increased cell viability and BPIFB4 expression, but decreased cell membrane pores, IL-1β and IL-18 levels in cell culture supernatant(t=3.827~10.919), and the mRNA and protein expression levels of NLRP3, ASC, as-pase-1, IL-1β, IL-18 and GSDMD mRNA and GSDMD-N protein (tmRNA=8.102~10.362,tprotein=8.647~10.947), the difference were statistically significant (all P<0.05). Compared with CSE+BPIFB4 group, CSE+BPIFB4+Nigericin sodium salt group ex-hibited decreased cell viability and BPIFB4 expression, but increased cell membrane pores, IL-1β and IL-18 levels in cell cul-ture supernatant(t=3.308~8.930), and the mRNA and protein levels of NLRP3, ASC, Caspase-1, IL-1β, IL-18 mRNA and pro-tein and GSDMD mRNA and GSDMD-N protein were elevated(tmRNA=5.759~9.413,tprotein=3.843~15.702), the difference were statistically significant (t=3.308~15.702, all P<0.05). Conclusions BPIFB4 can improve CSE-induced pyroptosis in BEAS-2B cells by inhibiting NLRP3 expression. Conclusion BPIFB4 can improve CSE-induced pyroptosis in BEAS-2B cells by inhibiting NLRP3 expression.

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备注/Memo

备注/Memo:
基金项目:新疆维吾尔自治区自然科学基金资助项目(2022D01F11);国家自然科学基金资助项目(82260013)。
作者简介:郑爱芳(1982-),女,本科,主治医师,研究方向:呼吸内科疾病,E-mail:bzy669@163.com。
通讯作者:李黎(1982-),女,硕士,主任医师,研究方向:呼吸内科疾病,E-mail:10517753@qq.com。
更新日期/Last Update: 2026-03-15