| 黄芩素经细胞焦亡途径调控支气管上皮炎性细胞因子的机制 |
| 投稿时间：2025-05-08 修订日期：2025-09-10 点此下载全文 |
| 引用本文：卢春亚,桑倩,林剑泷,李靖,屠蒙,王华启.黄芩素经细胞焦亡途径调控支气管上皮炎性细胞因子的机制[J].医学研究杂志,2025,54(10):40-44, 57 |
| DOI：
10.11969/j.issn.1673-548X.2025.10.008 |
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| 基金项目:国家自然科学基金资助项目(81972182);河南省医学科技攻关项目(LHGJ20230244) |
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| 中文摘要:目的 探讨黄芩素对支气管上皮细胞(bronchial epithelial cell,BEC)炎性细胞因子表达的影响以及调控机制。方法 取对数期16 HBE细胞,分为正常对照组、甲苯二异氰酸酯(toluene diisocyanate,TDI)刺激组及黄芩素低、中、高剂量组。正常对照组用正常培养基处理,TDI刺激组用TDI-人血清白蛋白(human serum albumin,HSA)刺激模拟哮喘细胞模型,黄芩素低、中、高剂量组经TDI-HSA刺激后分别用终浓度为10、20、40μmol/L的黄芩素继续干预48h。CCK-8法检测细胞活力；乳酸脱氢酶(lactate dehydrogenase,LDH)释放实验检测LDH释放率；流式细胞术检测各组焦亡率；酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测上清液白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-18(interleukin-18,IL-18)水平；免疫印迹(Western blot)法检测含NLR家族Pyrin域蛋白3(NLR family, pyrin domain containing protein 3,NLRP3)、半胱天冬酶-1(cysteine-dependent aspartate-specific protease 1,caspase-1)、Gasdermin D及凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)蛋白表达。结果 与正常对照组比较,TDI刺激组细胞活力降低(P＜0.05),LDH释放率、细胞焦亡率及上清液IL-1β、IL-6、IL-18水平均升高(P＜0.05)；与TDI刺激组比较,黄芩素低、中、高剂量组细胞活力升高(P＜0.05),LDH释放率、细胞焦亡率及上清液IL-1β、IL-6、IL-18水平均降低(P＜0.05),且表现为剂量依赖性(P＜0.05)。与正常对照组比较,TDI刺激组细胞NLRP3、caspase-1、Gasdermin D、ASC蛋白表达均升高(P＜0.05)；与TDI刺激组比较,黄芩素低、中、高剂量组细胞NLRP3、caspase-1、Gasdermin D、ASC蛋白表达均降低(P＜0.05),且表现为剂量依赖性(P＜0.05)。结论 黄芩素可能通过细胞焦亡途径减轻TDI诱导的支气管上皮细胞炎性细胞因子表达,提高细胞活力,降低细胞焦亡率。 |
| 中文关键词:哮喘 黄芩素 细胞焦亡 炎性细胞因子 |
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| Mechanism of Baicalein in Modulating Bronchial Epithelial Inflammatory Factors via the Pyroptosis Pathway. |
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| Abstract:Objective To investigate the effects of baicalein on the expression of inflammatory factors in bronchial epithelial cell (BEC), and explore the underlying regulatory mechanisms involved. Methods Log-phase 16 HBE cells were categorized into five groups:the normal control group, the toluene diisocyanate (TDI)-stimulated group, and the low-, medium-, and high-dose baicalein groups. The normal control group was cultured with standard medium, while the TDI-stimulated group was treated with TDI-human serum albumin (TDI-HSA) to establish an asthma-like cell model. Following TDI-HSA stimulation, the low-, medium-, and high-dose baicalein groups were intervened with baicalein at final concentrations of 10,20 and 40μmol/L for 48hours. Cell viability was assessed using the CCK-8 assay, and lactate dehydrogenase (LDH) release assays were performed to evaluate the LDH release rate. Flow cytometry was used to determine the rate of pyroptosis. Enzyme-linked immunosorbent assays (ELISA) were conducted to measure the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and interleukin-18 (IL-18) in the culture supernatants. Protein expression levels of NLR family, pyrin domain-containing protein 3 (NLRP3), cysteine-dependent aspartate-specific protease 1 (caspase-1), Gasdermin D and apoptosis-associated speck-like protein containing a CARD (ASC) were examined via Western blot analysis. Results Compared with the normal control group, the cell viability in the TDI stimulation group decreased (P<0.05), and the LDH release rate, pyroptosis rate, and the levels of IL-1β, IL-6 and IL-18 in the supernatant all increased (P<0.05). Compared with the TDI-stimulated group, the cell viability in the low-, medium-, and high-dose baicalein groups increased (P<0.05), while the LDH release rate, pyroptosis rate, and the levels of IL-1β, IL-6 and IL-18 in the supernatant all decreased (P<0.05), and all in a dose-dependent manner (P<0.05). Compared with the normal control group, the protein expressions of NLRP3, caspase-1, Gasdermin D and ASC in the TDI-stimulated group were all increased (P<0.05). Compared with the TDI-stimulated group, the protein expressions of NLRP3, caspase-1, Gasdermin D and ASC in the low-, medium- and high-dose baicalein groups were all decreased (P<0.05), also in a dose-dependent manner (P<0.05).Conclusion Baicalein may alleviate TDI-induced the expression of inflammatory factors in BEC by modulating the pyroptosis pathway. Through this mechanism, scutellarin enhances cell viability and reduces the rate of pyroptosis. |
| keywords:Asthma Baicalein Pyroptosis Inflammatory factor |
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