Study On The Protective Effect Of Bound Phenolics From Lycopus Lucidus Turcz. On Hepatocyte Injury In Vitro

Objective To determine the phenolic compositions of bound phenolic extracts from Lycopus lucidus Turcz.harvested from different sites at different harvest times.Through in vitro experiments,the protective effect of bound phenolic extracts on CCl₄ mediated the BRL rat hepatocyte damage was investigated.And its protective mechanism was preliminarily discussed,which provided experimental basis and theoretical basis for further revealing the functional activity of bound phenolic and the development and utilization of Lycopus lucidus Turcz..Methods Bound phenolic extracts from Lycopus lucidus Turcz.was obtained by alkaline extraction and X-5 macroporous resin was used for purification.The phenolic contents were determined by Folin-Ciocalteu method and the bound phenolic extracts composition was analyzed by high-performance liquid chromatography?HPLC?.In vitro CCl₄-induced hepatocyte injury model of BRL rats,by detecting liver function indicators alanine aminotransferase?ALT?,aspartate aminotransferase?AST?and lactate dehydrogenase?LDH?,inflammatory response factor tumor necrosis factor-??TNF-??,interleukin-6?IL-6?and interleukin-8?IL-8?levels,as well as apoptosis and Caspase-3 activity,to explore the effect of bound phenolic extracts from Lycopus lucidus Turcz.on hepatocyte injury induced by CCl₄ in vitro protection and mechanism.Results The phenolic contents of bound phenolic extracts from Lycopus lucidus Turcz.were in the range of 155.55?164.95?g GAE/mg EDW.The results of the analysis of monomer composition showed that the main phenolic compounds in the bound phenolic extracts were coffee acid?40.67?72.18?g/mg EDW?.The effects of coffee acid and rosemary acid on the content of bound phenolic extracts from Lycopus lucidus Turcz.were different in harvest period.The results of in vitro experiments showed that the concentration range of bound phenolic extracts from Lycopus lucidus Turcz.was 0.2?1.6 mg/mL,which had no effect on hepatocyte value-added activity in BRL rats.In the Liver cell injury protection system of BRL rats induced by CCl₄,the cell survival rate was 43.75?72.82 per cent when the concentration range of bound phenolic extracts from Lycopus lucidus Turcz.was 0.2?1.6 mg/mL,and the survival rate of the cells increased with the increase of the concentration.In the concentration range of 0.4?1.6 mg/mL,compared with the model group,bound phenolic extracts from Lycopus lucidus Turcz.significantly inhibited the increase of ALT,AST and LDH on hepatocyte injury induced by CCl₄ in BRL rats?p<0.05?,reduce the levels of inflammatory factors TNF-?,IL-6,IL-8 and inhibit apoptosis and Caspase-3 activation?p<0.05?.Conclusions The effects of sampling harvest times on the phenolic contents and compositions of bound phenolic extracts from Lycopus lucidus Turcz.were significantly different.The abundant phenolic compound in bound phenolic extracts was identified as caffeic acid.Significant protective effects of bound phenolic extracts from Lycopus lucidus Turcz.were observed on the CCl₄ mediated the BRL rat hepatocyte damage,and showed dose-effect dependence.The protective mechanism of bound phenolic extracts from Lycopus lucidus Turcz.on hepatocyte injury induced by CCl₄ in BRL rats was associated with inhibition of inflammatory response,inhibit apoptosis and Caspase-3 activation.