Expression Pattern And Regulation Of Adipocytes Differentiation Key Gene Apelin

The study of adipose tissue,not only help to improve the quality of animal products,but also can provide certain theoretical basis for obesity,diabetes,hypertension and other diseases.Adipose tissue is not only can store energy,but also can secrete adipocytokines and hormone,such as Apelin,PPAR?,leptin,resistin,etc.They regulate the metabolism of adipose tissue and adipocyte differentiation through autocrine or paracrine modes of action.It also can take part in the pathological process of adiposity,so it may play an important role during occurrence of some diseases such as obesity and insulin resistance.Apelin is an endogenous ligand for the APJ receptor.In2005,mRNA and protein of Apelin was being found in subcutaneous adipose tissue of rats and human.So some studies suggested that Apelin is an obesity-associated adipocytokine,it can increase the overall energy consumption and improve insulin sensitivity to suppress the occurrence of insulin resistance,and it may play an important role during occurrence of some diseases such as obesity and insulin resistance.Recent studies have indicated that Berberine can reduce the obesity mice weight by AMPK pathway improve glucolipid metabolism in mice.A number of studies show that berberine can inhibit the differentiation of adipocyde and the expression of PPAR? and C/EBP?.However,the molecular mechanism for the berberine regulation of adipocyte differentiation remains unclear.In this paper,we take the rats and pigs as research object.We use bioinformatics to realize the basic information about gene sequences and spatial structure of rat Apelin,for establishing the evolutionary tree and estimating the genetic relationship.By using RT-PCR,we explore the expression between organizations.At the same time for studying the differentially expressed among species,we compare the Apelin gene expression of two different rat and porcine.Using different concentrations of berberine to healthy SD rats by lavage,take the subcutaneous adipose tissue and visceral adipose tissue,to observe the mRNA and protein expression changes of Apelin,PPAR?,LPL,LXR? by RT-PCR and western blot.Finally try to know the underlying molecular mechanisms of the berberine in lipid-lowering.The main results were as follows:1.Porcine adipose tissue total RNA reverse transcription of cDNA first chain as a template,using reverse transcription PCR and other techniques to clone the pig's Apelin gene fragment,the length of the fragment 267 bp.The sequence of Apelin gene was obtained from the study of the differences in tissue expression of porcine,which provided the basis for genetic material and theory.2.The mRNA of the Apelin gene in seven tissues(heart,liver,spleen,lung,kidney,Muscle,adipose)from the pig and SD rat sample were examined by RT-PCR.The results showed that the Apelin gene was expressed specificity.The Apelin gene were expressed in heart,liver,spleen,lung,kidney and muscle tissue,and highly expressed in adipose tissue.While remarkable lower in heart,lung,spleen and muscle.SD rat and pig has the same expression regulation,and similarities between species.The results suggested that Apelin play an important role in lipid metabolism.3.Using different concentrations of berberine to lavage the SD rats.The results showed that with the berberine concentration increased,the level of transcription LPL,LXR? decreased significantly.Contrary,the level of transcription of Apelin showed a significant rise.The express of PPAR? genes was be inhibited in mRNA and protein expression,suggesting that berberine can slow down the formation of adipocyte.4.Protein structure analysis suggested that Apelin protein receptor is a transmembrane protein.The secondary structure of Apelin protein has more random curl which can form stability disulfide bond.The results show that the protein has signal peptide.Apelin protein is mainly composed of three functional domains: a low complexity of component area and a transmembrane region,a total of 22 amino acid from 5 to 27 residues constitute a transmembrane domain structure,and rat Apelin is a close relationship with people.Through the porcine Apelin gene cloning and of Apelin gene differentially expressions were studied,in order to increase the lean meat rate,improve pork quality and its application to provide a theoretical basis.Apelin gene has inhibitory effect on lipogenesis and berberine may by promoting the expression of Apelin gene in the body,decrease the amount of the expression of PPAR gamma gene and thus to lipid-lowering effect,the research on the fat synthesis and decomposition to provide new ideas for and improve the quality of human life has great significance.