竹节人参皂苷对乙醇致肝细胞L-O2损伤的保护作用

Protective effect of saponins extracted from Panax japonicus on ethanol-induced hepatic cells L-O2 injury

目的考察竹节人参皂苷对乙醇损伤肝细胞L-O2的保护作用,并探讨其作用机制。方法高效液相色谱-蒸发光散射检测法(HPLC-ELSD)分离纯化竹节人参皂苷,MTT法测定细胞存活率,分光光度法测定肝细胞内液丙二醛(MDA)含量、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活性。结果经分离纯化鉴定获得了6个竹节人参皂苷单体Rg1,Re,Rf,F3,Rg2和Rd;其中Rg10.16 g·L-1,Re 1.28g.L-1和Rf0.64 g·L-1可促进正常肝细胞L-O2增殖,增殖率分别为22.7%,34.8%和28.5%(P<0.01);Rd0.16 g·L-1和F31.28 g·L-1对正常肝细胞生长表现出明显的抑制作用,抑制率分别为49.7%和43.3%(P<0.01)。Rg10.16 g·L-1,Re 1.28 g·L-1和Rf 0.64 g·L-1对乙醇200 mmol·L-1损伤的肝细胞L-O2具有明显的保护作用,对细胞生长的抑制率由乙醇损伤模型组的50.4%分别降低为23.3%,26.9%和26.6%(P<0.01);Rd 0.16 g·L-1和F31.28 g·L-1则加强乙醇对肝细胞的损伤,抑制率高达83.2%和64.8%(P<0.01)。乙醇损伤模型组肝细胞L-O2乙醇代谢产生MDA含量升高、SOD和GSH-Px降低;Rg10.16 g·L-1,Re 1.28 g·L-1和Rf 0.64 g·L-1可明显改善乙醇损伤导致的肝细胞L-O2内MDA含量升高,增强SOD和GSH-Px活性(P<0.01)。结论人参皂苷Rg1,Re和Rf对乙醇损伤肝细胞L-O2具有明显的保护作用,抗氧化活性可能是其发挥保护作用的机制之一。

OBJECTIVE To study the protective effect of saponins from Panax japonicus against ethanol-induced hepatic cells L-O2 injury and their possible mechanisms.METHODSSaponins from P.japonicus was purified by HPLC-evaporation light scattering detection（ELSD）.The survival rate of hepatic cells was determined by MTT assay.The level of malondialdehyde（MDA）,the activities of superoxide dismutase（SOD） and glutathione peroxidase（GSH-Px） in L-O2 cells were measured by spectrophotometer.RESULTS Six major saponins from P.japonicus were identified by HPLC-ESI-MS： ginsenosides Rg1,Re,Rf,F3,Rg2 and Rd.Rg1 0.16 g·L^-1,Re 1.28 g·L^-1 and Rf 0.64 g·L^-1 could promote normal cells L-O2 proliferation（P〈0.01）,at the rate of 22.7%,34.8% and 28.5%,respectively.Rd 0.16 g·L^-1 and F3 1.28 g·L^-1 showed evident inhibition to normal cells L-O2（P〈0.01）,at the rate of 49.7% and 43.3%.Rg1 0.16 g·L^-1,Re 1.28 g·L^-1 and Rf 0.64 g·L^-1 also showed significantly protection against ethanol-induced hepatic cells L-O2 injury（P〈0.01）.The rate of inhibition decreased to 23.3%,26.9% and 26.6% compared with the ethanol-induced model control,the inhibition rate of which was 50.4%.Rd 0.16 g·L^-1 and F3 1.28 g·L^-1 showed more evident toxicity to ethanol-induced hepatic cells L-O2.The inhibition rate was 83.2% and 64.8%（P〈0.01）,respectively.Meanwhile,the content of MDA and the activity of SOD and GSH-Px in ethanol-induced L-O2 cells were（1.35±0.05）mmol·L^-1,（26.4±2.8）kU·L^-1 and（67.0±4.3）kU·L^-1,respectively.Rg1 0.16 g·L^-1,Re 1.28 g·L^-1 and Rf 0.64 g·L^-1 could decrease the level of intracellular MDA to（1.17±0.04）,（1.21±0.03） and（1.21±0.05）mmol·L^-1（P〈0.01）,increase the activity of intracellular SOD and GSH-Px activity to 38.3±4.8,35.6±4.4 and（39.0±3.3）kU·L^-1（P〈0.01）,and 86.3±5.7,78.4±3.5 and（84.2±4.4）kU·L^-1（P〈0.01）,respectively.CONCLUSIONRg1,Re and Rf have evidently protective effect against ethanol-induced hepatic cells L-O2 injury.The possible mechanism may be related to the decline of MDA content and increase in antioxidant enzymes such as SOD and GSH-Px activities.