Evaluation of chemopreventive effect of Fumaria indica against N-nitrosodiethylamine and CCl_4-induced hepatocellular carcinoma in Wistar rats

Objective:To investigation the chemopreventive potential of Fumaria indica（F.indica） extract（FIE） on N-nitrosodiethylamine and CCl4-induced hepatocarcinogenesis in Wislar rats.Methods:The experimental animals were divided into six groups（n=6）.Hepatocellular carcinoma was induced by single intraperitoneal injection of N-nitrosodiethylamine（NDEA） in normal saline at a dose of 200 mg/kg body weight followed by weekly subcutaneous injections of CCl4（3 mL/kg/week） for 6 weeks,as the promoter of carcinogenic effect.After administration of the carcinogen,200 and 400 mg/kg of FIE were administered orally once a day throughout the study.At the end of 20 weeks,the body weight,liver weight and relative liver weight were measured.The percentage of nodule incidence and liver cancer markers such as aspartate transaminase（AST）,alanine transaminase（ALT）,alkaline phosphatase（ALP）,γ-glutamyl transferase（γ-GT）,total bilirubin level（TBL）,α-feto protein（AFP） and carcinoembryonic antigen were estimated along with histopathological investigation in experimental groups of rats. Results:Obtained results demonstrated that the cotreatment with FIE significantly prevented the decrease of the body weight and also increased in relative liver weight caused by NDEA. The treatment with FIE significantly reduced the nodule incidence and nodule multiplicity in the rats after NDEA administration.The levels of liver cancer markers such as AST,ALT,ALP,γ-glutamyl transferase,TBL,AFP and carcinoembryonic antigen were substantially increased by NDEA treatment.However,FIE treatment significantly reduced the liver injury and restored the entire liver cancer markers.Histological observations of liver tissues too correlated with the biochemical observations.Conclusions:These finding powerfully supports that F.indica exert chemopreventive effect by suppressing the tumor burden and restoring the activities of hepatic cancer marker enzymes on NDEA and CCl4-induced hepatocarcinogenesis in Wistar rats.

Phytochemical investigation and simultaneous estimation of bioactive lupeol and stigmasterol in Abutilon indicum by validated HPTLC method

Objective:To perform a simultaneous quantitative estimation of two biologically active triterpenoid compounds lupeol and a steroid compound,stigmasterol,in Abutilon indicum(A.indicum)using high-performance thin-layer chromatography(HPTLC).Methods:TLC aluminum plates precoated with silica-gel 60 F254(20 cmí10 cm)were used with a mobile phase of toluene-methanol-formic acid(7.0:2.7:0.3,v/v/v)and densitometric determination of these compounds was carried out at 530 nm in reflectance/absorbance mode.Results:Compact bands for lupeol and stigmasterol were obtained at R_(f)0.52±0.02 and 0.28±0.05.The limit of detection(45 and 18 ng/band),limit of quantification(135 and 54 ng/band),recovery(98.2%-99.7%and 97.2%-99.6%)and precision(≤2.18 and 1.91)were satisfactory for lupeol and stigmasterol respectively.Linearity range for lupeol and stigmasterol were 100-1000(r^(2)=0.9994)and 50-500 ng/band(r^(2)=0.9941)and the contents were estimated as(0.59±0.10)%and(0.83±0.10)%w/w respectively.The total phenolic,flavonoid,proanthocyanidin,alkaloidal and saponin contents of methanolic extract of A.indicum were also measured in this work.According to International Conference on Harmonization(ICH)guidelines,the method was validated for linearity,precision,accuracy,and recovery,limit of detection,limit of quantification,specificity,and robustness.Conclusions:The HPTLC method was found to be reproducible,accurate,and precise and could detect these two compounds at nanogram level from the A.indicum.