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沈乐乐, 陈邦弟, 席建元.基于NLRP3/Caspase-1通路探讨银屑平丸含药血清对TNF-α诱导的人表皮角质形成细胞焦亡的影响[J].湖南中医药大学学报英文版,2024,44(10):1787-1793.[Click to copy
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| 基于NLRP3/Caspase-1通路探讨银屑平丸含药血清对TNF-α诱导的人表皮角质形成细胞焦亡的影响 |
| 沈乐乐,陈邦弟,席建元 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 探讨银屑平丸含药血清通过调控NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）/胱天蛋白酶-1（cysteine aspartate-specific proteinase-1，Caspase-1）信号通路改善肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）诱导的人表皮角质形成细胞焦亡的作用机制。方法 制备银屑平丸含药血清、阿维A含药血清及空白血清，采用CCK-8试剂检测空白血清对人表皮角质形成细胞存活率的影响；体外常规培养人表皮角质形成细胞，使用TNF-α诱导人表皮角质形成细胞建立体外银屑病损伤模型，设置正常组、模型组、空白血清组、银屑平丸含药血清组、阿维A含药血清组；ELISA 法检测各组细胞上清液中白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-18（interleukin-18，IL-18）水平；RT-PCR检测各组细胞NLRP3，功能蛋白D（gasdermin D，GSDMD）、Caspase-1 mRNA表达；Western blot 法检测各组细胞NLRP3、GSDMD、Caspase-1蛋白的表达。结果 与正常组比较，空白血清干预后细胞存活率无明显改变。与正常组比较，模型组和空白血清组细胞上清中IL-1β、IL-18水平显著升高（P＜0.01），且NLRP3、GSDMD、Caspase-1蛋白及mRNA表达水平显著升高（P＜0.05）；与模型组比较，银屑平丸含药血清组及阿维A含药血清组细胞上清IL-1β、IL-18蛋白及mRNA水平明显下降（P＜0.01），NLRP3、GSDMD、Caspase-1蛋白及mRNA表达显著下降（P＜0.01）。结论 银屑平丸含药血清可下调TNF-α诱导的人表皮角质形成细胞NLRP3、GSDMD、Caspase-1蛋白及mRNA表达，降低炎症因子表达水平，其作用机制可能与调控 NLRP3/Caspase-1通路有关。 |
| 关键词: 银屑病 银屑平丸含药血清 NLRP3/Caspase-1通路 人表皮角质形成细胞 细胞焦亡 |
| DOI：10.3969/j.issn.1674-070X.2024.10.010 |
| Received:February 22, 2024 |
| 基金项目:湖南省自然科学基金项目(2023JJ60490);湖南中医药大学科研项目(2022YYZK005);长沙市科技计划项目(Kq2202457)。 |
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| Effects of serum containing Yinxieping Pill on TNF-α-induced pyroptosis of human epidermal keratinocytes based on the NLRP3/Caspase-1 pathway |
| SHEN Lele, Chen Bangdi, XI Jianyuan |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To investigate the mechanism of action of serum containing Yinxieping Pill (SCYXPP) in improving pyroptosis of human epidermal keratinocytes induced by tumor necrosis factor-α (TNF-α) through regulating the NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/cysteine aspartate-specific proteinase-1 (Caspase-1) signaling pathway. Methods SCYXPP, serum containing Acitretin (SCA), and blank serum were prepared. CCK-8 reagent was used to detect the effec of blank serum on cell viability of the human epidermal keratinocytes. Human epidermal keratinocytes were cultured in vitro under conventional conditions and TNF-α was applied to them to establish an in vitro psoriasis injury model. Then normal, model, blank serum, SCYXPP, and SCA groups were set up. The levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the cell supernatant of each group were determined by ELISA, the mRNA expressions of NLRP3, gasdermin D (GSDMD), and Caspase-1 in the cells of each group were examined by RT-PCR, and the protein expressions of NLRP3, GSDMD, and Caspase-1 were checked by Western blot. Results Compared with the normal group, there was no significant change in cell viability in the blank serum group. Compared with the normal group, the levels of IL-1β and IL-18 in the cell supernatant of the model and blank serum groups were significantly elevated (P<0.01), and the mRNA and protein expressions of NLRP3, GSDMD, and Caspase-1 were significantly higher (P<0.05). Compared with the model group, the IL-1β and IL-18 levels in the cellular supernatant of SCYXPP and SCA groups significantly decreased (P<0.01), and the mRNA and protein expressions of NLRP3, GSDMD, and Caspase-1 were significantly reduced (P<0.01). Conclusion SCYXPP can down-regulate the mRNA and protein expressions of NLRP3, GSDMD, and Caspase-1 in human epidermal keratinocytes induced by TNF-α, and reduce the expression levels of inflammatory factors. Its mechanism of action may be related to the regulation of the NLRP3/Caspase-1 pathway. |
| Key words: psoriasis serum containing Yinxieping Pill NLRP3/Caspase-1 pathway human epidermal keratinocytes pyroptosis |
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