To investigate the cholesterol-lowering effects of alfalfa saponin extract (ASE) and its regulation mechanism on some key genes involved in cholesterol metabolism 40 healthy 7 weeks old male Sprague Dawley (SD) rats were randomly divided into four groups with 10 rats in each group: control group GANT 58 hyperlipidemic group ASE treatment group ASE prevention group. abnormal serum lipid levels in hyperlipidemic rats were ameliorated by ASE Rabbit Polyclonal to ARNT. administration (both ASE prevention group and treatment group) (and in the liver of hyperlipidemic rats were amazingly down-regulated (and were dramatically up-regulated by both ASE administration (and and in the liver of hyperlipidemic rats which was involved in cholesterol biosynthesis uptake and efflux pathway; (2) the increase in excretion of cholesterol. The findings in our study suggested ASE experienced great potential usefulness as a natural agent for treating hyperlipidemia. Introduction Hyperlipidemia is usually common in the worldwide population and is considered as a highly modifiable risk factor for cardiovascular disease such as coronary heart disease and peripheral artery diseases. Elevated blood lipid levels especially increased serum low-density lipoprotein (LDL) level can accelerate atherosclerosis. Therefore reducing high lipid levels has been considered to be an important approach to prevent or slow the progression of atherosclerosis [1]. In recent decades increasing attention has been paid to new natural brokers with lipid-reducing activity [2]-[4]. The cholesterol-lowering effects and prevention GANT 58 in cardiovascular disease of saponins have been exhibited [5] [6]. Alfalfa (and in hepatic tissues among treatments. was used as an internal control. The primers for QPCR were presented in Table 1. Real-time QPCR was performed on a 96-well PCR plate in triplicate with GANT 58 a total reaction volume of 10 μL made up of 1 μL cDNA 5 uL SYBRGreen Mastermix 0.1 uL of each specific forward GANT 58 and reverse primers and 3.8 μL nuclease-free water PCR was carried out in an ABI PRISM 7700 sequence detection system (Applied Biosystems) with 2 min at 95°C for predegeneration then followed by 40 cycles at 95°C for 15 s 60 for 20 s and 72°C for 30 s each. The reaction mixture with no cDNA was considered as the unfavorable control to confirm the absence of primer dimerization. The cycle threshold (Ct) values were determined by Sequence Detection System software version 1.7a. Qualitative PCR was performed to confirm formation of a single product in each reaction before quantitation. The target gene expressions of the samples were exhibited as fold change from control. All genes were normalised with ASE treatment group; ASE prevention group) but the administration of ASE experienced a trend to reduce body weight gain of hyperlipidemic rats although they did not reach the values of control group (both ASE administration groups). No significant differences in relative liver excess weight of rats among treatments were observed (both ASE administration groups) which indicated the beneficial effects of ASE on serum lipid profiles in hyperlipidemic rats. Physique 2 Effects of alfalfa saponin extract on serum lipid levels of rats. Effects of ASE on total cholesterol and total bile acids levels in liver and feces of rats As summarized in Physique 3 and ?and4 4 the rats fed with high-lipid diet showed markedly higher levels of liver TC and TBA compared with the control group (both ASE administration groups) however both ASE administration significantly increased liver TBA level (in liver of hyperlipidemic rats was down-regulated as compared with the control group (in liver of hyperlipidemic rats was up-regulated as compared with the control group (in liver of hyperlipidemic rats was up-regulated as compared to the control group (both ASE GANT 58 administration groups). On the contrary gene expression of in liver of hyperlipidemic rats was down-regulated as compared to the control group (both ASE administration groups). Compared with ASE treatment group gene expression of and of rats in ASE prevention group were significantly increased (was suppressed in hyperlipidemic rats and further significantly inhibited by ASE administration and markedly reduced TC level in the liver and serum of hyperlipidemic rats. So the decreasing TC level in the liver and serum observed in the current study could be explained by the down-regulation of mRNA levels and.