紫檀芪对脂多糖诱导的人HL-7702肝脏细胞损伤作用及机制

Effects and mechanisms of pterostilbene on lpopolysaccharide-induced liver injury of HL-7702

目的探索紫檀芪(pterostilbene,PTE)对细菌脂多糖(lipopolysaccharide,LPS)诱导的人HL-7702肝脏细胞损伤的作用及机制。方法建立LPS诱导的人HL-7702细胞炎症损伤模型,分别使用不同浓度(1μg/ml、5μg/ml、10μg/ml、20μg/ml、40μg/ml、60μg/ml、80μg/ml、100μg/ml,6 h)的LPS诱导肝细胞损伤,选择合适的作用浓度。分别使用10μg/ml、15μg/ml、20μg/ml的PTE预处理HL-7702模型细胞。利用MTT法检测各组细胞存活率。利用Western blot检测各组细胞的B淋巴细胞瘤-2(B-cell lymphoma-2,BcL-2)、B淋巴细胞瘤-2相关的蛋白质X(BcL2-asssociated X,Bax)、核因子-κB(nuclear factor kappa-B,NF-κB)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)的蛋白表达水平。结果与正常对照组相比,100μg/ml LPS损伤组细胞A值显著下降(0.54±0.01 vs 0.38±0.02),Bax(0.36±0.07 vs 0.87±0.09)、NF-κB(0.40±0.01 vs 0.90±0.02)、TNF-α(0.35±0.07 vs 0.90±0.04)及IL-6(0.30±0.04 vs 0.73±0.09)蛋白表达水平显著上升,BcL-2蛋白表达下调(0.81±0.08 vs 0.32±0.07),差异均具有统计学意义(P均<0.05)。与100μg/ml LPS组细胞相比,15μg/ml PTE预处理组A值显著增加(0.34±0.01 vs 0.46±0.01)、Bax(0.87±0.09 vs 0.61±0.09)、NF-κB(0.90±0.02 vs 0.73±0.06)、TNF-α(0.90±0.04 vs 0.66±0.06)及IL-6(0.73±0.09 vs 0.53±0.03)蛋白表达水平显著下降,BcL-2蛋白表达上调(0.32±0.07 vs 0.68±0.02),差异均有统计学意义(P均<0.05)。结论PTE可抑制LPS诱导的肝细胞凋亡,减少炎性介质产生,其机制可能与抑制NF-κB信号转导通路的激活有关。

Objective To evaluate the effects and mechanisms of pterostilbene(PTE)on lipopolysaccharide(LPS)-induced liver injury of HL-7702.Methods HL-7702 cells were stimulated with different concentration(1μg/ml,5μg/ml,10μg/ml,20μg/ml,40μg/ml,60μg/ml,80μg/ml,100μg/ml)of LPS for 6 h to select the appropriate concentration in order to establish the inflammation injury model.HL-7702 cells were treated with PTE(10μg/ml,15μg/ml,20μg/ml)after inflammation injury model was established.MTT assay was performed to detect the cell viability and the levels of B-cell lymphoma-2(BcL-2),BCL2-associated X(Bax),nuclear factor kappa-B(NF-κB),tumor necrosis factor-α(TNF-α)and interleukin-6 were evaluated by Western blot.Results Compared with blank control group,A values of 100μg/ml LPS group decreased significantly(0.54±0.01 vs 0.38±0.02),while Bax(0.36±0.07 vs 0.87±0.09),NF-κB(0.40±0.01 vs 0.90±0.02),TNF-α(0.35±0.07 vs 0.90±0.04)and IL-6(0.30±0.04 vs 0.73±0.09)levels increased significantly,BcL-2 level decreased significantly,the differences were statistically significant(all P<0.05).Compared with 100μg/ml LPS group,A values of 15μg/ml PTE group increased significantly(0.34±0.01 vs 0.46±0.01),while Bax(0.87±0.09 vs 0.61±0.09),NF-κB(0.90±0.02 vs 0.73±0.06),TNF-α(0.90±0.04 vs 0.66±0.06)and IL-6(0.73±0.09 vs 0.53±0.03)levels decreased significantly,BcL-2 level increased significantly,the differences were statistically significant(all P<0.05).Conclusions PTE alleviates LPS-induced HL-7702 cells injury by reducing inflammatory response and inhibiting cell apoptosis,the mechanism may be related to the inhibition of the activation of NF-κB signaling transduction pathway.