Effect of alcohol on induction of differentiation of NIH 3T3 fibroblast into adipocyte / 中国组织工程研究

ABSTRACT

OBJECTIVE: Animal experiments prove that alcohol may cause adipo-accumulation in femoral head and then induce osteonecrosis,but its mechanism of action is not known. NIH 3T3 fibroblast is a kind of pluripotent stem cell.There are not relative reports on the effect of alcohol on NIH 3T3fibroblast at present.OBJECTIVE: To observe the effect of alcohol on the differentiation of NIH 3T3 fibroblast into adipocyte and the influence of that on the expression of adipogenic transcription factor-peroxisome proliferator activated receptor gamma (PPAR-γ).DESIGN: Single sample observation.SETTING: Department of Biochemistry, Basic Medical College,Zhengzhou University and Department of Orthopaedics, First Affiliated Hospital, Zhengzhou University.MATERIALS The experiment was completed in the Laboratory of Department of Biochemistry, Basic Medical College, Zhengzhou University from April 2002 to June 2003.NIH 3T3 fibroblasts were provided by the Laboratory of Department of Orthopaedics,University of Virginia Hospital.METHODS: The NIH 3T3 fibroblasts were divided into 6 groups randomly.The alcohols (volume fraction was 0.998) with the dosage 0(control group),0.03,0.06,0.09,0.15,0.21 mol/L were added respectively to the NIH 3T3 fibroblasts daily.The mixtures were cultured for 14 days and then stained with Sudan Ⅳ .The percentage of adipocyte was determined with image analysis software of computer,Image Pro Plus 4.1.The expression of PPAR-γ mRNA was determined with the technology of reverse transcription-polymerase chain reaction.alcohol and control groups.RESULTS: The cells were processed with the alcohols of progressive inpercentages of adipocytes in all the alcohol groups were 11.9% ,23.8%,28.2% ,31.1% ,42.6% respectively and 1.2,2.5,2.9,3.2,4.4 times that in control group(9.6%) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant (P ＞ 0.05),but the differences between the other 4 groups and control group,0.03 mol/L alcohol ues of PPAR-γ mRNA to 18S gene in the cells of all the groups were 0.218,0.411,0.486,0.473,0.453 respectively and were 1.1,2.1,2.5,2.4,2.3times that in control group (0.197) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant(P ＞ 0.05),but the differences between the other 4 groups and control group and 003 mol/L alcohol group were significant extremely(P ＜ 0.001).CONCLUSION: Alcohol can induce differentiations of a large quantity of NIH 3T3 fibroblasts into adipocytes directly, which might be one of the factors of lipotrophy in bone marrow wheri alcohol-induced osteonecrosis happens.