Study Of Alfalfa Saponins Extract On Cholesterol Metabolism Gene Expression Regulation In Laying Hens

This study was conducted to explore effect of adding different levels alfalfa saponins extract on cholesterol metabolism enzyme gene mRNA expression, by measuring the changes in gene expression of mRNA which is related to cholesterol metabolism.Through the Gel shift Assay of alfalfa saponins extract reguLate the gene expression of cholesterol metabolism, to explore the mechanism of the alfalfa saponins reduce the expression of cholesterol metabolism gene. The resuLts as follows:Exp.1:Effect of alfalfa saponins extract on the gene expression of cholesterol metabolism of laying hens.Alfalfa saponins was applicated in the diet of laying hens.150 hailanhe laying hens which was 190-days-old were randomLy divided into 5 treatment groups with 5 replicates and 6 per replicates. The experiment lasted 77 days. The control group was fed with the basal diets, The treatment groups were fed with 60 mg/kg、120 mg/kg、 240 mg/kg、480 mg/kg alfalfa saponins respectively adding to the basal diets. The resuLts showed that:(1) The mRNA expression of Vldl receptor was suppressed due to adding alfalfa saponins extract supplementation in the diet.(2) The mRNA expression of Cyp7a1 were significantly upreguLated.(3) Alfalfa saponins extract did not exert significance effect on the cholesterol gene of apoVLDL-Ⅱ、SREBP-2、HMGCR、 ApoB100、LDLR.Exp.2:The exploration of experiment of alfalfa saponins extract reguLate the cholesterol metabolism expression of laying hens.In order to investigate the alfalfa saponins extract effec the expression of cholesterol metabolism genes were reguLated directly or indirectly.The liver of laying hens which used in this experiment were from the Exp.1.The HMGCR gene, asa example, according to the different mobility of nucleic acid and protein by EMSA, identify the transcription factor binding proteins which binding to the nucleic acid probe. The resuLts showed that:(1) the gene of HMGCR was successfuLly cloned upstream non-coding region 2000bp fragment, while the probe divided into fragments between 100-200bp. (2)The nuclear protein werer successfuLly extracted which will bind the probe in the experiment through incubation then forming complexes. (3) The conditions of EMS A parameter was optimized through the experiment.