| It is called Angiopoietin-like protein 6 (Angptl 6/AGF) for its molecular structure and the angiopoietin amino acid sequence is very similar. Angptl 6 can inhibit the gluconeogenesis pathway to prevent the deposition of fatty liver, thereby regulating lipid metabolism. Resveratrol is a stilbene structure containing the natural polyphenols, resveratrol has anti-aging effect and the anti-cancer, anti-inflammatory, reduces blood glucose and cardiovascular protection, have a certain effect. High dose (3-5g) of resveratrol can increase energy consumption, lower blood sugar. Baicalin baicalin is in a class of flavonoids, baicalin has antibacterial,diuretic, anti-inflamm atory, anti-metamorphosis, solutions spasm, anti-platelet aggregation, lowering blood pressure, treatment of cardiovascular disease, liver or gallbladder cancer reactions Effect. Magnolol honokiol extract of Chinese medicine is the main active substance Magnolia. Honokiol have lower cholesterol, low density lipoprotein oxide production and the formation of atherosclerosis.Firstly,according to the Angptl6 gene sequences of cattle, pigs, horses, two pairs of specific primers were designed and synthesized., we cloned total RNA from goat liver by molecular biological methods and got Angptl 6 gene fragment. Primary cultured cells extracted from goat liver were stimulated by three kinds of medicinal plants monomer in 3 different concentrations (40μM,20μM and 10μM) of resveratrol, baicalin and magnolol.Using Angptl 6 gene as a target, we measured the expression of Angptl6 mRNA by quantitative PCR. It is expected to explore the kinds of metabolic mechanisms of these three aspects of drug regulation in a view of genetic point. The results are as follows:1. With structural integrity, total RNA extracted from goat liver holds its high purity. The size of the specificity amplified fragment tallies with the expected ones, and the sequencing results have uploaded to Genbank.2,Isolated by collagenase double perfusion, the total amount of goat liver cells are achieved to (2.065±0.485)×108 (n=4), and the yield is 1.032×107/g in each liver tissue. With 0.4% trypan blue dye test, immediate survival rate of the goat liver cells is (86.42± 2.57%). However, the morphology of liver cell has structural integrity and good viability. The first five days of high LDH levels up to 87 U/L in the Supernatant. Five days later, the self-repairing of goat liver cells decreased gradually LDH to a smooth level, about to 51 U/L, which indicates the healthy growth of the cells at this time in good condition with their membrane integrity. Supernatant BUN concentration curve has been steady. Cultured cells can maintain a good blood of urea nitrogen synthesis. In the first five days, the concentration of ALB has been increasing with high cell albumin secretion, which began to decline in Sixth day and then leveled off. The synthesis of albumin in hepatocytes gradually becomes stable. Compared with perfusion, tissue culture method is much more simple, but the success rate is low, the growth rate of cells is slower, and there are much more other tissue fragments like the pre-cultured blood cells.3,the fluorescence quantitative PCR melting curve is a single peak. All resveratrol, baicalin and magnolol can make goat liver cell mRNA expression increased. |
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