ABSTRACT
The specific expression of TGH and HSL genes in different tissues of Bamei pig was investigated by RT-PCR and Western blot in this study. The result of RT-PCR showed that the expression of HSL could be detected in all these seven tissues examined, and which was higher expressed in fat, lower in heart, liver, lung, spleen and kidney. Expression of TGH gene could also be detected in seven tissues, and higher in liver and fat, lower in heart and kidney and lowest in spleen and lung. The result of Western blot showed that, HSL gene was highest expressed in epiploica fat and subcutaneous fat, higher in other tissues, but couldn' t be detected in kidney. Expression of TGH was detected in epiploica fat, subcutaneous fat, liver, lung and spleen, and highest in fat and liver, but it hadn't be found in heart and kidney. These results suggested that both HSL and TGH could be regulated by post-transcriptional, and their function was involved in different tissues.
Subject(s)
Animals , Male , Gene Expression Regulation , Lipase , Genetics , Metabolism , Sterol Esterase , Genetics , Metabolism , Swine , Tissue DistributionABSTRACT
<p><b>OBJECTIVE</b>To study the effect of emodin (EMO) on the proliferation and differentiation of rat preadipocytes.</p><p><b>METHOD</b>Separating and culturing rat preadipocytes, grouping the wells that preadipocytes were growing according to certain concentration such as 0, 5, 10, 20, 40, 80, 160 micromol x L(-1) randomly, MTT spectrophotometry and flow cytometry (FCM) were adopted to determine the effect of EMO on proliferation of rat preadipocytes. The accumulation of TG (triglyceride) in adipocytes was assayed by oil red O staining, and the morphological changes of the adipocytes were determined by morphology observation.</p><p><b>RESULT</b>EMO in the range of 20-160 micromol x L(-1) could inhibit the proliferation and differentiation of preadipocytes in a dose and time dependent manner, and induce apoptosis of preadipocytes in a certain degree.</p><p><b>CONCLUSION</b>EMO should have a potential to serve as a fat-reducing drug.</p>
Subject(s)
Animals , Male , Rats , Adipocytes , Cell Biology , Apoptosis , Cell Cycle , Cell Differentiation , Cell Proliferation , Cells, Cultured , Dose-Response Relationship, Drug , Emodin , Pharmacology , Random Allocation , Rats, Sprague-DawleyABSTRACT
Total RNA was isolated from kidney of BaMei pig, a local strain of Chinese pig, and then the cDNA sequence of SOCS-2 gene was cloned by RT-PCR (GenBank accepted number is EF121242). Then the cloned SOCS-2 gene was inserted into PMD19-T vector by T/A cloning, transformed into DH-5alpha, tested by PCR and sequenced. The data show that the homology of the cloned porcine SOCS-2, including 822 bp, is more than 93% and that of the deduced amino acid sequence is 89% when compared with human, rat and mice. And the molecular weight of SOCS-2 protein is about 22.25 kD and PI is 8.03. The cloning of SOCS-2 gene is useful for the further research on the molecular mechanism by which regulating growth and development of organism.
Subject(s)
Animals , Humans , Rats , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA, Complementary , Molecular Sequence Data , Protein Structure, Secondary , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis , Sequence Homology , Suppressor of Cytokine Signaling Proteins , Genetics , Swine , GeneticsABSTRACT
1 approximately 3 days old Piglet's primary preadipocytes in vitro were cultured and treated with 0micromol/L (control group), 10microlmol/L (lower dose group), 20micromol/L(middle dose group) and 50micromol/L, 100micromol/L (higher dose group) RES. Cell proliferation and viability were analyzed by MTT assay. The degree of differentiation and adipogenesis were measured by Oil Red O staining extraction assay and the expression of Sirt1 (sirtuin) mRNA were detected by RT-PCR. The results showed the optical density (OD) of MTT and Oil Red O staining were all decreased, especially treated by 50micromol/L, 100micromol/L RES at 72h and 96h (P < 0.01); the ratio of OD of the expression of Sirt1 mRNA to that of beta-actin mRNA were increased after treated by 100micromol/L RES (P < 0.01). RES can inhibit proliferation and differentiation of pig preadipocytes in certain degree. Higher dose of RES can markedly decrease adipogenesis and prevent preadipocytes differentiation into adipocytes, which may be in part associated with its effect on increasing the expression of Sirt1 mRNA.
Subject(s)
Animals , Adipocytes , Cell Biology , Metabolism , Cell Differentiation , Cell Proliferation , Cells, Cultured , RNA, Messenger , Sirtuin 1 , Genetics , Stem Cells , Stilbenes , Pharmacology , Swine , Transcription, GeneticABSTRACT
To investigate the effects of Baicalein (BAI) on the proliferation and differentiation of pig preadipocytes, and elucidate its potential mechanism. Primary preadipocytes of pig were cultured in vitro. The morphologic changes of preadipocytes differentiation were observed by Oil Red O staining. Status of cell proliferation was detected by MTT assay. The degree of adipogenesis and differentiation were measured by Oil Red O staining extraction assay. The activity of fatty acid synthase (FAS) was detected by spectrophotometry. The mRNA expression of special peroxisome proliferation activated receptor-gamma2 gene (PPARgamma2) was detected by reverse transcriptase polymerase chain reaction (RT-PCR). When preadipocytes differentiated into adipocytes, the preadipocytes were changed from shuttle shape to oval or round, in which big and small lipid droplets were filled. The proliferation of preadipocytes was inhibited by the treatment of 160-640 micromol/L BAI (P < 0.05). The mRNA expression of PPARgamma2 and FAS activity and the differentiation of preadipocytes was repressed by 40-320 micromol/L BAI treatment (P < 0.05). It is concluded that the proliferation and differentiation of preadipocytes is inhibited by BAI in some degree. The effect of BAI on differentiation of preadipocytes may be resulted from inhibiting the mRNA expression of PPARgamma2 and reducing FAS activity.
Subject(s)
Animals , Adipocytes , Cell Biology , Metabolism , Azo Compounds , Metabolism , Cell Differentiation , Cell Proliferation , Dose-Response Relationship, Drug , Flavanones , Pharmacology , Gene Expression Regulation , Obesity , Pathology , PPAR gamma , Genetics , RNA, Messenger , Genetics , Metabolism , Swine , fas Receptor , MetabolismABSTRACT
To investigate effects of docosahexaenoic acid (DHA) on proliferation and differentiation of rat adipocytes and to elucidate its potential mechanism, rat's primary preadipocytes in vitro were cultured. Treated adipocytes with 0 micromol/L (control group), 40 micromol/L (lower dose group) and 160 micromol/L (higher dose group) DHA. Cell living rations and proliferation were analyzed by trypan blue exclusion and MTT assay. The degree of adipogenesis and differentiation were measured by Oil Red O staining extraction assay and the expression of peroxisome proliferation activated receptor-gamma2 (PPARgamma2) mRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). It was demonstrated that cells living ration and the optical density (OD) of MTT were all decreased, especially treated by 160 micromol/L DHA at 60 (72 hours (P < 0.05). The OD of Oil Red O staining and the expression of PPARgamma2 mRNA were all decreased after treated by 160 micromol/L DHA (P < 0.01). It can be concluded that DHA can inhibite proliferation and differentiation of adipocytes in some degree. Higher dose of DHA can markedly decrease adipogenesis and prevent differentiation of adipocytes, which may be in part associated with its effect on decreasing the expression of PPARgamma2 mRNA.