烧伤血清对3T3-L1前脂肪细胞增殖活性和成脂分化能力的影响  

INFLUENCE OF BURN SERUM ON PROLIFERATION AND ADIPOSE DIFFERENTIATION OF 3T3-L1 PREADIPOCYTES

作　　者：庄淑波[1,2] 柴家科[1] 郁永辉[1] 段红杰[1] 刘玲英[1] 樊军[1] 侯玉森 王一贺 

机构地区：[1]解放军总医院第一附属医院烧伤整形外科,北京100048 [2]解放军医学院

出　　处：《中国修复重建外科杂志》2014年第6期763-767,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81120108014;81071544;81372052);首都医学发展科研基金资助项目(2009-2040);National Natural Science Foundation of China(81120108014;81071544;81372052);Capital Medical Development Scientific Research Funds(2009-2040)

摘　　要：目的通过观察大鼠烧伤血清对3T3-L1前脂肪细胞增殖活性和成脂分化能力的影响,探索烧伤对脂肪代谢的影响。方法选择成年雄性SD大鼠48只,随机分为假伤组及烧伤1、4、7、14、21 d组,每组8只。各烧伤组大鼠背部制备30%总体表面积的Ⅲ度烫伤模型,分别于对应时间点收集血清;假伤组不作处理,取血清作为正常对照。采用各组血清培养3T3-Ll前脂肪细胞,MTT法检测细胞增殖活性,选择增殖能力最低组血清作为后续刺激血清。取3T3-Ll前脂肪细胞分别采用假伤血清(A组)、烧伤血清(B组)、假伤血清成脂诱导(C组)、烧伤血清成脂诱导(D组)培养。倒置显微镜下观察细胞生长情况,诱导培养7 d后行油红O染色,进行脂肪细胞计数并测量吸光度(A)值,实时定量PCR及Western blot检测过氧化物酶体增殖物激活受体γ(preoxisome proliferator-activated receptorγ,PPAR-γ)和脂蛋白脂酶(lipoprotein lipase,LPL)基因及蛋白表达。结果 MTT检测示,与其余各组比较,烧伤4、7 d组细胞增殖能力显著降低(P<0.05),其中7 d组最低(P<0.05),取烧伤7 d组大鼠血清作为后续刺激血清。倒置显微镜观察示,培养后A、B组细胞形态无明显变化;诱导培养后C、D组细胞呈脂肪细胞样变,其中C组最显著。油红O染色示,C组细胞染色呈阳性,D组为弱阳性,余均为阴性;脂肪细胞计数以及A值比较示,A组高于B组、C组高于D组,差异均有统计学意义(P<0.05)。实时定量PCR及Western blot检测示,B组PPAR-γ基因相对表达量明显低于A组(P<0.05),但LPL基因相对表达量差异无统计学意义(P>0.05);PPAR-γ、LPL蛋白表达量比较差异无统计学意义(P>0.05)。D组PPAR-γ、LPL基因及蛋白表达量均显著优于C组(P<0.05)。结论大鼠烧伤7 d血清可致3T3-Ll前脂肪细胞成脂分化能力下降。Objective To investigate the effect of burn on the fat metabolism by observing the effect of burn serum on the proliferation and adipose differentiation of 3T3-L1 preadipocytes. Methods Forty-eight male Sprague Dawley rats were randomly divided into sham burn group and burn at 1, 4, 7, 14, and 21 days groups, 8 rats in each group. The rats in burn groups were made the full-thickness thermal burns comprising 30% total body surface area. At 1, 4, 7, 14, and 21 days after burn, the serum of burn rats was collected. The rats in sham burn group were not treated as normal control. The proliferation activity of 3T3-L1 cells was detected using MTT method after treated by normal and burn serum. The burn serum having the highest proliferation inhibitory effect was chosen for subsequent study. The growth of 3T3-L1 cells in normal serum group （group A）, burn serum group （group B）, normal serum and adipogenic induction group （group C）, burn serum and adipogenic induction group （group D） was observed using inverted microscope. After 7 days of treatment, the adipocytes was stained by oil red O and the absorbance （A） value was measured. The mRNA and protein levels of preoxisome proliferator-activated receptor y （PPAR-Y） and lipoprotein lipase （LPL） were detected by real-time quantitative PCR and Western blot. Results The proliferation ability of 3T3-L1 cells was significantly reduced in the group treated by 4- or 7-day burn serum （P 〈 0.05）, especially 7-day burn serum treatment group （P 〈 0.05）. Under inverted microscope, the cell morphology in group A and group B had no obvious change, but a large number of fat ceils were observed in group C and a few were observed in group D. The positive or weak positive oil red O staining was observed in group C or group D, respectively. The cell counting and A value were significantly higher in group A than in group B, and in group C than in group D （P 〈 0.05）. The mRNA level of PPAR-y in group B was significantly reduced when compared with tha

关 键 词：烧伤血清 3T3-Ll前脂肪细胞 成脂分化 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]