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目的探讨灰树花多糖(PGF)对四氯化碳诱导的肝L-02细胞损伤的保护作用。方法培养人肝L-02细胞,建立CCl4肝细胞损伤模型,分组实验:正常对照组、CCl4损伤组和不同浓度的PGF(50、100、200和400μg/mL)保护组。采用形态学观察,MTT检测,生化分析培养液中ALT、AST活性及细胞内SOD活性、MDA含量,流式细胞仪检测线粒体膜电位、荧光探针观测胞内Ca2+浓度;Western blot检测细胞bcl-2、bax蛋白表达。结果与CCl4损伤组相比,各PGF保护组细胞活力明显增强,上清液中ALT、AST活性明显降低;细胞内SOD明显升高、Ca2+浓度MDA含量降低,细胞bcl-2的表达上调、bax的表达下调。以100μg/mL浓度保护效果最佳。结论 PGF对CCl4诱导的肝L-02细胞损伤有明显的保护作用,其机制可能与清除自由基、抑制肝细胞内Ca2+浓度的升高、改变凋亡相关蛋白bcl-2、bax的表达水平,有效地防止线粒体膜电位的下降、减少细胞凋亡有关。
Objective To investigate the protective effect of Grifola frond polysaccharide (PGF) on carbon tetrachloride-induced liver L-02 cell injury. Methods Human liver L-02 cells were cultured to establish CCl4 hepatocytes injury model. The rats were divided into normal control group, CCl4-injured group and different concentrations of PGF (50, 100, 200 and 400 μg / mL). Morphological observation, MTT assay, biochemical analysis of ALT, AST activity and intracellular SOD activity, MDA content, mitochondrial membrane potential measured by flow cytometry, intracellular Ca2 + concentration by fluorescent probe; Western blot detection of bcl- 2, bax protein expression. Results Compared with CCl4-injured group, the viability of all PGF-treated groups was significantly increased, and the activities of ALT and AST in supernatant were significantly decreased. The intracellular SOD was significantly increased, the content of Ca2 + in MDA was decreased, the expression of bcl-2 was up-regulated and bax Down-regulated 100μg / mL concentration of the best protection. Conclusion PGF can protect CCl4-induced liver L-02 cells from injury. Its mechanism may be related to the scavenging of free radicals, inhibiting the increase of Ca2 + concentration in hepatocytes, changing the expression levels of bcl-2 and bax, Effectively prevent the mitochondrial membrane potential decline, reduce apoptosis.