南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (05): 669-.

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骨髓源性肥大细胞对软骨细胞表达Ⅱ型胶原及糖胺多糖的影响

欧阳晴晴,赵进军,杨敏   

  • 出版日期:2014-05-20 发布日期:2014-05-20

Effects of bone marrow-derived mast cells on expressions of type II collagen and
glycosaminoglycan in co-cultured chondrocytes

  • Online:2014-05-20 Published:2014-05-20

摘要: 目的探讨骨髓源性肥大细胞(BMMCs)对软骨细胞表达Ⅱ型胶原和糖胺多糖(GAG)的影响。方法原代培养小鼠
BMMCs和关节软骨细胞,将第2代软骨细胞以及培养4周的BMMCs,按照1∶10的比例种植于Transwell共培养系统中。细胞
分为4个组:软骨细胞组(对照组)、软骨细胞+BMMCs组(BMMCs组)、软骨细胞+BMMCs+色甘酸二钠组(DSCG组)、软骨细
胞+BMMCs+Compound48/80组(C48/80组),每组设3个复孔,收集软骨细胞并裂解,随后ELISA以及Western Blotting检测软
骨细胞内的Ⅱ型胶原的表达,用1,9-二甲基亚甲蓝法(DBM)检测软骨细胞内GAG的含量。结果在培养24 h,BMMCs组和
DSCG组Ⅱ型胶原以及GAG的含量与对照组间无差异(P>0.05),C48/80组Ⅱ型胶原与GAG含量相对于对照组和BMMCs组显
著降低(P<0.01),48 h的结果与24 h一致,24 h与48 h之间比较,各组之间均无差异(P>0.05)。结论C48/80激活的BMMCs可
降低软骨细胞Ⅱ型胶原以及GAG表达。

Abstract: Objective To investigate the influence of the bone marrow-derived mast cells (BMMCs) on the expression of type II
collagen and glycosaminoglycan (GAG) in chondrocytes co-cultured with BMMCs. Methods Primarily cultured mouse
BMMCs at 4 weeks and the second passage of chondrocytes were plated in a Transwell co-cultured system at a ratio of 1∶10 in
the presence or absence of sodium cromoglycate (DSCG) or compound 48/80 (C48/80). The chondrocytes were harvested and
lysed for detecting type II collagen expression with ELISA and Western blotting and GAG expression using 1,9
dimethylmethylene blue (DBM). Results After a 24-hour culture, the chondrocytes co-cultured with BMMCs showed similar
expression levels of type II collagen and GAG to the control group regardless of the presence of DSCG (P>0.05). Compared
with chondrocytes cultured alone or with BMMCs, the co-cultured chondrocytes in the presence of C48/80 showed
significantly lower expressions of type II collagen and GAG (P<0.01). Such results did not vary significantly as the culture time
was extended to 48 h. Conclusions C48/80-activated BMMCs can reduce the expression of type II collagen and GAG in
chondrocytes in the co-culture system.