The Determination Of Chemical Constituents In Trgionella Foenumgraecum L And Therapeutic Effects Of TFGe On Alcoholic Fatty Liver Of Rats

Objective:To establish a method for the quantitiative determination ofTrigonelline and diosgenin in different extract of Fenugreek by HPLC. Antioxidantactivity on DPPH and hydroxyl freebase (OH) with different extract of Fenugreek werestudied in this paper. To observe therapeutic effects of Trigonella foenum greacum.Lextract of alcohol （TFGe）on alcoholic fatty liver(AFL)of rats，trying to clarify themechanism of it’s therapeutic effects.Method:Using the CHP method to determinateTrigonelline; diosgenin was analyzed by HPLC on Hypersil ODS-1columns(4.6×250mm，5μm) with the mobile phase of methanol acetonitrile-water(86:14,v/v),the flow rate1.0mL/min,detection wavelength203nm,column temperature35°C. Vitro antioxidant effects were studied by UV method. The model of alcoholic fattyliver was established by intragastricly feeding rats with the mixture of white spirit andemulsion containing high fat for10weeks. When pathological inspection showed that themodel of alcoholic fatty liver of rats was established，intragastricly fed rats with TFGe for4weeks，at the same time，the positive control group was intragastricly fed with silymarinand TFGe. After the rats were killed，liver homogenate was prepared. Serum level of totalcholesterol(TC)、 triglyceride(TG)、 high density lipoprotein(HDL-C)、 low densitylipoprotein(LDL-C)、total bilirubin (TBili)、albumin(ALB) was detected: activity ofAspartate aminotransferase (AST)、 alanine aminotransferase (ATL)、glutamyltranspeptidase (GGT) was detected; hepatic contents of TC、TG were tested:Serum level of tumor necrosis factor(TNF-α)、adiponectin and AMPK was examined byenzyme-linked immunospecific assay (ELISA)method.Results:The liner range ofdiosgenin was43.2μg/ml~864μg/ml (r=0.9995); average recovery (n=9) was100.3%.Results showed that: Fenugreek extract have Scavenging effects on DPPH and hydroxylradicals (OH). With TFGe treatment，contents of serum TG、TC、LDL-C of AFL ratswere significantly decreased; activity of AST、ATL was depressed; hepatic contents of TC、TG were significantly reduced: the level of Serum TNF-α decreased while level ofadiponectin increased highly. Steatosis of hepatic tissue obviously improved. But Serumlevel of TG increased as compared with that of the normal group.the level of Serum TNF-αdecreased while level of adiponectin、SOD、AMPK increased highly.Conclusion:TheHPLC method is accurate and reliable, good reproducibility, and could be used in thequantitative determination of Fenugreek extract. Fenugreek extract have effects onScavenging DPPH and hydroxyl radicals.TFGs have therapeutic effects alcoholic fattyliver of rats including:reduce blood fat， decrease liver fat， decrease activity oftransaminase.