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The Effects Of Berberine On Human Adipocytes Secretoring Adiponectin And Leptin

Posted on:2008-07-31Degree:MasterType:Thesis
Country:ChinaCandidate:J P CaoFull Text:PDF
GTID:2144360215988465Subject:Internal Medicine
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Object:1.To culture human preadipocytes primarily and induce differentiation to adipocytes.2.To study the effects of berberine on human adipocytes secretoring adiponectin and leptin.Method:1.Cell culture of human preadipocytes:Omental adipose tissue biopsies were obtained from patients undergoing elective open-abdominal surgery.None of the patients had diabetes or severe systemic illness or had any recent weight change.None were taking medications known to affect adipose tissue mass or metabolism.Age of patients was below 50,and body mass index was below 25.Adipose tissue was separated from omental adipose tissue and capillaries by dissection and then digested with collagenase type 1.The digested tissue was filtered through nylon membrance.the filtrate was centrifugated at 600xg for 10 min and discarded supernatant and re-suspended in the DMEM/F12 basal medium.The cells were counted and planted.2.Induce preadipocytes differentiation to adipocytes:After planting for 12-hour at 37℃under 5%CO2 atmosphere,most of the cell have adhered.Changing the basal medium into the differentiational medium.The first three days,Making use of the differentiational medium with 3-isobutyl-1-methylxanthine.From the fourth day to eighth day,utilizating the differentiational medium without 3-isobutyl-1-methylxanthine.At the ninth day,the cells were divided into control group and berberine group and incubated for 24 hours.To collecte cell supernatant before and after intervention and store at -80℃.Preparing for assaying adiponectin and leptin at the same time3.Assay concentration of culture solution:Concentration of culture solution adiponectin and leptin were assayed by double- antibody enzyme linked immunosorbent assay(BA-ELISA)。4.Statistical analysis:All statistical analysis was performed with Windows SPSS13.0.Date are presented as (?)±s.The differences between the groups were determined by t-test.Differences were considered significant at a value of P<0.05.Results:1.To culture successfully human preadipocytes primarily.We can cultivate the preadipocytes of uniform components by the way of collagenase digestion.2.To induce human preadipocytes differentiation to adipocyte.In vitro,the human preadipocytes can be induced to be adipocytes under the effect of differentiation inducing factor.3.After 24-hour adipocytes intervented with 10μmol/l berberine,the change before and after adiponectin and leptin excretion is respective(0.99±0.06 VS 0.62±0.03)and(187.13±5.56 VS 137.62±4.31).Conclusion:1.To culture successfully human preadipocytes primarily and induce differentiation to adipocytes.It provides experimental cell.model for studying of endocrinal function of adipocytes that culturing human adipocytes.2.Ex vivo,berberine decreases human adipocytes secretory adiponectin and leptin.To study the direcrt effect of berberine to adipocytes in vitro,which establishes the experimental foundation for identifying the mechanism of improving Insulin resistance.
Keywords/Search Tags:Berberine, Adiponectin, Leptin, Adipocytes
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