CAJ | 학술논문

根据形态学变化,甘油-3-磷酸脱氢酶(GPDH)活力的升高和三酸甘油酯(TG)积累的增加与胚牛血清(EBS)相比,猫血清能显著地使元代培养中大鼠前脂细胞发生分化作用,GPDH酶活力因加猫血清者为:373+/-45单位/mg蛋白质,而加入FBS者为:118+/-23单位/mg蛋白质;N＝21,P<0.001,相对应的TG分别为:16.1+/-2.7μmol/mg DNA与5.5+/-1.2μmol/mg DNA,N＝12,P<0.0005.分化细胞引发的脂肪细胞转化作用,GPDH:FBA与胰岛素组为1247+/-82单位/mg蛋白质,而猫血清+FBS+胰岛素组为1145+/-80单位/mg蛋白质.猫血清还对大鼠前脂细胞具有促进有丝分裂的作用,尤其在接种后的第4天至第5天最为明显.此种促分化作用成分在56℃经45分钟后仍稳定,但经100℃30分钟处理即遭破坏.它是非透析性的,对胰蛋白酶、链霉菌蛋白酶和羧肽酶A只有抗性,但胰凝乳蛋白酶却可使其部分地失活.它对DTT和高碘酸盐不敏感,在pH2和pH12条件下不稳定,其等电点为5左右,它能与Con A琼脂糖相结合,看来是一种糖蛋白.凝胶过滤层析表明:它的分子量为57KDa.结论:猫血清含有一种能促使元代培养中的大鼠前脂细胞转化成脂肪细胞,但却没有使3T3细胞系细胞发生这种作用,表明这两种细胞存在着固有的差异.
근거형태학변화,감유-3-린산탈경매(GPDH)활력적승고화삼산감유지(TG)적루적증가여배우혈청(EBS)상비,묘혈청능현저지사원대배양중대서전지세포발생분화작용,GPDH매활력인가묘혈청자위:373+/-45단위/mg단백질,이가입FBS자위:118+/-23단위/mg단백질;N＝21,P<0.001,상대응적TG분별위:16.1+/-2.7μmol/mg DNA여5.5+/-1.2μmol/mg DNA,N＝12,P<0.0005.분화세포인발적지방세포전화작용,GPDH:FBA여이도소조위1247+/-82단위/mg단백질,이묘혈청+FBS+이도소조위1145+/-80단위/mg단백질.묘혈청환대대서전지세포구유촉진유사분렬적작용,우기재접충후적제4천지제5천최위명현.차충촉분화작용성분재56℃경45분종후잉은정,단경100℃30분종처리즉조파배.타시비투석성적,대이단백매、련매균단백매화최태매A지유항성,단이응유단백매각가사기부분지실활.타대DTT화고전산염불민감,재pH2화pH12조건하불은정,기등전점위5좌우,타능여Con A경지당상결합,간래시일충당단백.응효과려층석표명:타적분자량위57KDa.결론:묘혈청함유일충능촉사원대배양중적대서전지세포전화성지방세포,단각몰유사3T3세포계세포발생저충작용,표명저량충세포존재착고유적차이.
Differential responsiveness of rat preadipocytes and 3T3-F442A cells to maturation induction was studied in the presence of cat serum. As judged by morphological change and increases in glycerol-3-phosphate dehydrogenase(GPDH)activity and triacylglycerol(TG) accumulation, cat serum induced significantly differentiation of preadipocytes in primary culture as compared to fetal bovine serum(FBS)(GPDH: 373±45 vs.118±23 U/mg protein with FBS,n=21, p<0.001;TG:16.1±2.7 vs 5.8±1.2 μmol/mg DNA for FBS, n=12, p<0.0005).The differentiation-promoting effect is a dose -dependent fashion. In contrast,cat serum neither promoted nor inhibited appreciable adipose conversion of 3T3-F442A cells by FBS and insulin(GPDH:1247+82 vs.1145±80 U/mg protein with cat serum). The pronouced mitogenic effect of cat serum on rat preadipocytes was also observed, especially from day 4 to day 5 post plating. The adipogenic principle(s) is stable at 56℃ for 45 min but destroyed at 100℃ for 30 min, non-dialyzable, resistant to trypsin, pronase and carboxypeptidase A but partially inactivated by chymotrypsin, insensitive to DTT and periodate, unstable at pH 2 and pH 12 and has a pI of about 5. It is a glycoprotein binding to ConA-Argarose. Gel filtration chromatography showed that the differentiation activity corresponded to a molecular weight of 57 kDa. We conclude that there is a principle(s) in cat serum which promotes adipose conversion of rat preadipocytes in primary culture, but not for 3T3 clonal cells, and that inherent differences exist between rat preadipocytes and 3T3 cell lines.