华支睾吸虫排泄/分泌产物对肝纤维化的影响
Effect of excretory/secretory products of Clonorchis sinensis on liver fibrosis
孙云异 1魏伟 1毛瑞锋 1张馨慧 1王佳文 1刘雪薇 1高俊峰 1王春仁1
作者信息
- 1. 黑龙江八一农垦大学 动物科技学院,黑龙江 大庆 163319
- 折叠
摘要
为了探讨华支睾吸虫排泄/分泌产物(ESPs)对大鼠肝星状细胞(HSCs)的影响,采用不同质量浓度的华支睾吸虫ESPs(CsESPs)处理HSC-T6 细胞,利用CCK8 法检测CsESPs对HSC-T6 细胞活力的影响,并以MCC950 预处理细胞作为对照,利用RT-qPCR和Western-blot分别检测细胞中NLRP3、Caspase-1、IL-1β、α-SMA、Col1α1 和Col3α1 的基因和蛋白表达水平.结果显示,与对照组相比,25、50 和 100 μg/mL的CsESPs对细胞活力无明显影响;当CsESPs的质量浓度达到 50 μg/mL时,与对照组相比,NLRP3 及其下游的Caspase-1、IL-1β基因的相对表达水平显著升高,HSC-T6 细胞激活的标志基因α-SMA及纤维化相关的Col1α1 基因的相对表达水平也显著升高;Western-blot结果显示,与对照组相比,50 μg/mL的CsESPs可以使 HSC-T6 细胞中NLRP3、Caspase-1、IL-1β、α-SMA、Col1α1 及Col3α1 的蛋白表达水平显著升高,而在MCC950 预处理细胞组,这 6 个指标的基因和蛋白表达水平均被明显抑制.结果表明,CsESPs 可以使HSC-T6 细胞活化,诱导细胞内NLRP3 炎性小体活化及胶原表达,NLRP3 炎性小体活化是华支睾吸虫感染所致肝纤维化的关键环节之一.
Abstract
In order to investigate the effect of Clonorchis sinensis excretory/secretory products(ESPs)on rat hepatic stellate cell(HSCs),the HSC-T6 cells were treated with different concentrations of CsESPs,and the effects of CsESPs on cell viability were detected by CCK8 method,and at the same time,MCC950 pretreated cells were used as a control,and RT-qPCR and Western-blot were used to detect the gene and protein expression levels of NLRP3,Caspase-1,IL-1β,α-SMA,Col1α1 and Col3α1 in HSC-T6 cells.The results showed that,compared to the control group,25,50 and 100 μg/mL CsESPs had no significant effect on cell viability.When the concentration of CsESPs reached 50 μg/mL,the relative gene expres-sion levels of NLRP3 and its downstream Caspase-1 and IL-1β were significantly increased compared to the control group,and the mRNA expression levels of α-SMA as the marker gene for HSC-T6 cells activa-tion,and Col1α1 as the fibrosis gene,were also significantly higher compared to the control group.Western-blot results showed that,compared to the control group,50 μg/mL CsESPs significantly in-creased the protein expression levels of NLRP3,Caspase-1,IL-1β,α-SMA,Col1α1 and Col3α1 in HSC-T6 cells.But at MCC950 pretreatment groups,the genes and protein expression levels of these six indica-tors were significantly inhibited.The results indicated that CsESPs can activate HSC-T6 cells and in-duce the activation of NLRP3 inflammasome and collagen expression in cells.The activation of NLRP3 in-flammasome was one of the key links of liver fibrosis caused by Clonorchis sinensis infection.
关键词
华支睾吸虫/排泄/分泌产物/肝星状细胞/NLRP3炎性小体/肝纤维化Key words
Clonorchis sinensis/excretory/secretory products/hepatic stellate cells/NLRP3 in-flammasome/liver fibrosis引用本文复制引用
基金项目
国家自然科学基金(31972703)
国家自然科学基金(32172886)
黑龙江省自然科学基金(ZD2022C006)
黑龙江八一农垦大学三横三纵支持计划(ZRCQC202204)
出版年
2024
国家科技期刊平台
NSTL
万方数据