片仔癀通过调控mir-155表达缓解LPS刺激肝脏Kupffer细胞的损伤

Pien Tze Huang alleviates LPS-stimulated liver Kupffer cell injury by regulating miR-155 expression

目的探究片仔癀通过调控miR-155表达改善脂多糖(lipopolysaccharide,LPS)诱导的肝脏Kupffer细胞损伤的发生机制。方法LPS诱导肝脏Kupffer细胞建立细胞损伤模型,模拟脓毒症肝损伤。RT-qPCR检测损伤细胞中miR-155的表达,RT-qPCR、Western Blot、ELISA和流式细胞术评估损伤细胞的炎症反应和细胞凋亡情况。然后用不同浓度(0 mg/L、5 mg/L、10 mg/L及15 mg/L)的片仔癀处理LPS诱导的Kupffer细胞,检测了细胞中miR-155的表达,细胞的炎症反应和凋亡率。在细胞损伤模型中沉默miR-155,RT-qPCR、Western Blot、ELISA和流式细胞术评估miR-155对模型细胞炎症反应和凋亡的影响。在片仔癀处理的损伤细胞中过表达miR-155检测细胞炎症反应和凋亡的变化。数据以均数±标准差(x±s)的表示,采用成组t检验或单因素方差分析各组数据。结果在LPS诱导肝脏Kupffer细胞损伤的模型中,miR-155的表达显著升高(P<0.05),促炎因子IL-6、TNF-α的表达水平显著增加,抗炎因子IL-10明显被抑制(P<0.05),细胞凋亡率显著升高(P<0.05)。片仔癀处理后,抑制损伤肝细胞中miR-155的表达(P<0.05),抑制细胞炎症因子IL-6、和TNF-α的水平,促进抗炎因子IL-10的表达(P<0.05),抑制细胞凋亡(P<0.05)。沉默miR-155减轻了细胞的炎症反应和凋亡率(P<0.05)。过表达miR-155可逆转片仔癀治疗肝细胞损伤的作用(P<0.05)。结论在LPS诱导肝脏Kupffer细胞损伤的模型中,片仔癀通过抑制miR-155的表达,抑制细胞的炎症反应及其凋亡。

Objective To explore the mechanism by which Pien Tze Huang improves liver Kupffer cell damage induced by lipopolysaccharide(LPS)by regulating the expression of miR-155.Methods LPS induced liver Kupffer cells to establish a cell injury model to simulate septic liver injury.RT-qPCR was used to detect the expression of miR-155 in damaged cells,and RT-qPCR,Western Blot,ELISA and fl ow cytometry were used to evaluate the infl ammatory response and apoptosis of damaged cells.Then we treated LPS-induced Kupffer cells with Pien Tze Huang at different concentrations(0 mg/L,5 mg/L,10 mg/L and 15 mg/L),and detected the expression of miR-155 in the cells,the infl ammatory response of the cells and Apoptosis rate.MiR-155 was silenced in the cell injury model,and RT-qPCR,Western Blot,ELISA and flow cytometry were used to evaluate the effect of miR-155 on inflammatory response and apoptosis of model cells.Overexpression of miR-155 in damaged cells treated with Pien Tze Huang was used to detect changes in cellular infl ammatory response and apoptosis.Data are expressed in the form of mean±standard deviation,and each group of data is analyzed using t test or one-way analysis of variance.Results In the LPS-induced liver Kupffer cell injury model,the expression of miR-155 was signifi cantly increased(P<0.05),the expression levels of pro-infl ammatory factors IL-6 and TNF-αwere signifi cantly increased,and the antiinfl ammatory factor IL-10 was signifi cantly increased.was inhibited(P<0.05),and the cell apoptosis rate was signifi cantly increased(P<0.05).After Pien Tze Huang treatment,the expression of miR-155 in damaged liver cells was inhibited(P<0.05),the levels of cellular infl ammatory factors IL-6 and TNF-αwere inhibited,and the expression of anti-infl ammatory factor IL-10 was promoted(P<0.05).Inhibit cell apoptosis(P<0.05).Silencing miR-155 reduced the infl ammatory response and apoptosis rate of cells(P<0.05).Overexpression of miR-155 can reverse the effect of Pien Tze Huang on liver cell injury(P<0.05).Conclusions In the model of LPS-induced liver Kupffer cell injury,Pien Tze Huang can inhibite the infl ammatory response and apoptosis of cells by inhibiting the expression of miR-155.