中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2009年
4期
268-271
,共4页
折涛%胡大海%张军%刘佳琦%张万福%蔡维霞%赵周婷%汤朝武
摺濤%鬍大海%張軍%劉佳琦%張萬福%蔡維霞%趙週婷%湯朝武
절도%호대해%장군%류가기%장만복%채유하%조주정%탕조무
胰岛素%干细胞%成纤维细胞%细胞增殖%胶原合成
胰島素%榦細胞%成纖維細胞%細胞增殖%膠原閤成
이도소%간세포%성섬유세포%세포증식%효원합성
Insulin%Stem cells%Fibrublasts%Cell proliferation%Collagen synthesis
目的 了解不同浓度胰岛素对脂肪干细胞(ADSC)分泌生长因子功能的影响.方法 获取人腹部脂肪组织分离培养ADSC,选择第3代细胞经免疫表型鉴定及脂源性诱导分化鉴定.根据培养液中胰岛素浓度,将ADSC分为1×10-8、1×10-7、1 × 10-6mol/L 3个实验组,常规培养至70%融合时,更换为含不同浓度胰岛素的无血清DMEM培养液培养3 d.设无胰岛素培养液培养的ADSC为对照组.采用酶联免疫吸附测定法,测定ADSC的血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)分泌量;应用噻唑蓝比色法和羟脯氨酸比色法,检测ADSC上清液对Fb增殖及胶原合成的影响. 结果 1×10-8、1×10-7mol/L胰岛素组ADSC的VEGF、HGF分泌量分别为(471±41)、(762±66)ng/L和(643±64)、(930±67)ng/L,与对照组(286±47)、(577±84)ng/L比较,其含量显著增加(P<0.05或P<0.01);而1×10-6mol/L胰岛素组ADSC的VEGF、HGF分泌量无明显变化(P>0.05).1×10-8、1×10-7moL/L胰岛素组ADSC培养上清液有明显的促Fb增殖及胶原合成作用,以1×10-7mol/L组作用更显著(P<0.05或P<0.01). 结论 1×10-8、1×10-7mol/L胰岛素能显著促进ADSC分泌VEGF和HGF.
目的 瞭解不同濃度胰島素對脂肪榦細胞(ADSC)分泌生長因子功能的影響.方法 穫取人腹部脂肪組織分離培養ADSC,選擇第3代細胞經免疫錶型鑒定及脂源性誘導分化鑒定.根據培養液中胰島素濃度,將ADSC分為1×10-8、1×10-7、1 × 10-6mol/L 3箇實驗組,常規培養至70%融閤時,更換為含不同濃度胰島素的無血清DMEM培養液培養3 d.設無胰島素培養液培養的ADSC為對照組.採用酶聯免疫吸附測定法,測定ADSC的血管內皮生長因子(VEGF)、肝細胞生長因子(HGF)分泌量;應用噻唑藍比色法和羥脯氨痠比色法,檢測ADSC上清液對Fb增殖及膠原閤成的影響. 結果 1×10-8、1×10-7mol/L胰島素組ADSC的VEGF、HGF分泌量分彆為(471±41)、(762±66)ng/L和(643±64)、(930±67)ng/L,與對照組(286±47)、(577±84)ng/L比較,其含量顯著增加(P<0.05或P<0.01);而1×10-6mol/L胰島素組ADSC的VEGF、HGF分泌量無明顯變化(P>0.05).1×10-8、1×10-7moL/L胰島素組ADSC培養上清液有明顯的促Fb增殖及膠原閤成作用,以1×10-7mol/L組作用更顯著(P<0.05或P<0.01). 結論 1×10-8、1×10-7mol/L胰島素能顯著促進ADSC分泌VEGF和HGF.
목적 료해불동농도이도소대지방간세포(ADSC)분비생장인자공능적영향.방법 획취인복부지방조직분리배양ADSC,선택제3대세포경면역표형감정급지원성유도분화감정.근거배양액중이도소농도,장ADSC분위1×10-8、1×10-7、1 × 10-6mol/L 3개실험조,상규배양지70%융합시,경환위함불동농도이도소적무혈청DMEM배양액배양3 d.설무이도소배양액배양적ADSC위대조조.채용매련면역흡부측정법,측정ADSC적혈관내피생장인자(VEGF)、간세포생장인자(HGF)분비량;응용새서람비색법화간포안산비색법,검측ADSC상청액대Fb증식급효원합성적영향. 결과 1×10-8、1×10-7mol/L이도소조ADSC적VEGF、HGF분비량분별위(471±41)、(762±66)ng/L화(643±64)、(930±67)ng/L,여대조조(286±47)、(577±84)ng/L비교,기함량현저증가(P<0.05혹P<0.01);이1×10-6mol/L이도소조ADSC적VEGF、HGF분비량무명현변화(P>0.05).1×10-8、1×10-7moL/L이도소조ADSC배양상청액유명현적촉Fb증식급효원합성작용,이1×10-7mol/L조작용경현저(P<0.05혹P<0.01). 결론 1×10-8、1×10-7mol/L이도소능현저촉진ADSC분비VEGF화HGF.
Objective To study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs). Methods ADSCs were isolated from human ab-dominal adipose tissue and cultured. The immunophenotype and adipose induced-differenciation were identi-fied, and the third generation cells were collected. The collected cells were assigned to 1 × 10-8, 1 × 10-7, 1 × 10-6 mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry. Results The secretion amounts of VEGF and HGF of ADSCs in 1 × 10-8 and 1 × 10-7 mol/L insulin groups [ (471±41, 762±66 ng/L), (643±64, 930±67 ng/L) , respectively ] were significantly higher as compared with those in control group (286±47, 577±84 ng/L) ( P <0.05 orP <0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1±10-6 mol/L insulin group ( P >0.05 ). The supernatant fluid of ADSCs' nutrient medium of 1 ± 10-8 ,1 ± 10-7 mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 ± 10-7 mol/L group ( P < 0.05 or P < 0. 01 ). Conclusions Insulin in the concentrations of 1 ± 10-8 and 1 ± 10-7 mol/L can notably promote ADSCs' function of secreting VEGF and HGF.
