CAJ | 학술논문

目的探讨体外脂肪源性干细胞(ADSCs)向软骨诱导的过程中,不同浓度白细胞介素-1β(IL-1β)对ADSCs向软骨细胞分化的影响. 方法体外成功分离、培养大鼠ADSCs,实验分4组(n=12):对照组、实验A、B、C组,每组加入等量的细胞数(1 ×105个),实验A、B、C组中分别加入浓度为1、2、3 ng/mL的IL-1β.培养6、12、18d应用实时定量聚合酶链式反应检测各组成软骨基因(Ⅱ型胶原和糖胺聚糖蛋白)和成脂基因(脂肪酸结合蛋白)的表达量. 结果 ADSCs免疫化学染色结果显示CD44和CD105呈阳性表达培养6、12、18 d,实验B组Ⅱ型胶原、糖胺聚糖蛋白基因表达量较对照组明显增加,差异均有统计学意义(P＜0.05);而实验A、C组之间及分别与对照组比较差异均无统计学意义(P＞0.05).培养6、12、18 d,实验A组脂肪酸结合蛋白基因表达量较对照组和实验B组明显增加,实验B组脂肪酸结合蛋白基因表达量较对照组明显减少,差异均有统计学意义(P＜0.05);而实验C组与对照组之间比较差异无统计学意义(D＞0.05). 结论当IL-1β浓度为1、3ng/mL时,其对ADSCs向软骨细胞分化未见明显促进作用;但当IL-1β浓度为2 ng/mL时,可对ADSCs的成软骨分化产生促进作用.
목적 탐토체외지방원성간세포(ADSCs)향연골유도적과정중,불동농도백세포개소-1β(IL-1β)대ADSCs향연골세포분화적영향. 방법 체외성공분리、배양대서ADSCs,실험분4조(n=12):대조조、실험A、B、C조,매조가입등량적세포수(1 ×105개),실험A、B、C조중분별가입농도위1、2、3 ng/mL적IL-1β.배양6、12、18d응용실시정량취합매련식반응검측각조성연골기인(Ⅱ형효원화당알취당단백)화성지기인(지방산결합단백)적표체량. 결과 ADSCs면역화학염색결과현시CD44화CD105정양성표체배양6、12、18 d,실험B조Ⅱ형효원、당알취당단백기인표체량교대조조명현증가,차이균유통계학의의(P＜0.05);이실험A、C조지간급분별여대조조비교차이균무통계학의의(P＞0.05).배양6、12、18 d,실험A조지방산결합단백기인표체량교대조조화실험B조명현증가,실험B조지방산결합단백기인표체량교대조조명현감소,차이균유통계학의의(P＜0.05);이실험C조여대조조지간비교차이무통계학의의(D＞0.05). 결론 당IL-1β농도위1、3ng/mL시,기대ADSCs향연골세포분화미견명현촉진작용;단당IL-1β농도위2 ng/mL시,가대ADSCs적성연골분화산생촉진작용.
Objective To investigate the effects of interleukin-1β (IL-1β) at different concentrations on the differentiation of adipose derived stem cells (ADSCs) into chondrocytes in vitro.Methods ADSCs were successfully isolated,cultured and identified in vitro from SD rats.According to the preliminary results,the experiments were performed in 4 groups (3 experimental and one control ones) with 12 samples in each.The same quantity of cells (1 × 105) was used in each group.IL-1β at concentrations of 1 ng/mL,2ng/mL and 3 ng/mL was added respectively in experimental groups A,B and C.At 6,12 and 18 days after culture,the gene expression levels of cartilage formation (type Ⅱ collagen and glycosaminoglycan protein) and lipoid formation (fatty acid binding protein) were detected using RT-PCR.Results Immunochemistry showed positive expression of CD44 and CD105 in ADSCs.At 6,12 and 18 days after culture,the expression levels of type Ⅱ collagen and glycosaminoglycan protein in group B were significantly increased than in the control group(P ＜ 0.05).There were no such significant differences between groups A and C,or between the control group and groups A and C respectively (P ＞ 0.05).At 6,12 and 18 days after culture,the expression of fatty acid binding protein was significantly higher in group A than in the control group and group B,and significantly lower in group B than in the control group(P ＜ 0.05),but there was no significant difference between group C and the control group (P ＞ 0.05).Conclusion IL-1 β at concentrations of 1 ng/mL and 3 ng/mL has no significant effect on the differentiation of ADSCs into chondrocytes in vitro,but IL-1β at a concentration of 2 ng/mL may have.