绞股蓝多糖含药血清对四氯化碳肝细胞损伤的保护作用

Protective effect of PGP-containing mouse serum on HepG2 cells injured by CCl_4 and its possible mechanism

目的研究绞股蓝多糖(gynostemma pentaphyllum makino polysaccharide,PGP)含药血清对四氯化碳(car-bon tetrachloride,CCl4)诱导的肝细胞损伤保护作用。方法采用中药血清学方法收集PGP含药血清,用CCl4诱导肝HepG2细胞损伤,设正常对照组、CCl4损伤组和不同浓度PGP含药血清保护组;细胞形态学观察细胞凋亡,MTT法检测细胞活力,生化法检测细胞培养上清液中丙氨酸氨基转移酶(alanine aminotransferase,ALT/GPT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST/GOT)活性及western blot凝胶电泳检测细胞色素P450 2E1的表达。结果 各PGP含药血清保护组肝细胞损伤程度明显减轻;细胞存活率显著升高(P<0.05);上清液中ALT、AST活性显著降低(P<0.05);CYP 2E1表达量显著增加。以4～8 mg·kg-1·d-1保护组效果最佳。结论 PGP含药血清对CCl4诱导的HepG2细胞损伤有明显保护作用,以4～8 mg·kg-1·d-1含药血清作用最显著。

Objective To investigate the protective effect of gynostemma pentaphylhim makino polysaccharide（ PGP）-containing mouse serum on HepG2 cells injured by a 70% carbon tetrachloride（ CC14 ） saturated medium. Methods PGP-containing mouse serum was collected from PGP-treated mice. HepG2 cells used in the experiment were divided into the following groups： the control group, the CC14 injury group and the PGP-containing mouse serum protection groups with different concentrations of PGP. A 70% CCI4 saturated medium was used to induce HepO2 cell injury in all groups except the control group. Cell morphological change was observed under a bright-field microscope, cell viability by 4-methyl- tetrazolium （M＇IT） chromatometry, the degree of cell damage by measuring supernatant alanine aminotransferase （GPT） and aspartate aminotransferase （GOT） activities, and expression level of the cytochrome P450 2E1 （ CYP2E1 ） protein by Western blot. Results Compared with the CC14 injury group, the PGP-containing serum protection groups had significant morphological improvement, and their survival rate was significantly higher （P 〈 0.05 ）. ALT and AST activities in the supernatant were significantly decreased （P 〈0.05） while expression level of CYP2E1 was increased.The protective effect was the most obvious in the 4～8mg·kg^-1·d^-1 PGP-containing serum protection group. Conclu- sion Pre-treatment of PGP-containing mouse serum is shown to possess a significant protective effect on HepG2 cells injured by CC14 especially at 4～8mg·kg^-1·d^-1.