摘要
目的利用膜片钳研究L型钙通道对软骨细胞合成Ⅱ型胶原的作用。方法分离培养兔关节软骨细胞，随机分为对照组和胰岛素样生长因子（IGF）组。采用膜片钳记录L型钙通道的变化；激光共聚焦显微镜观察1周时细胞内钙离子浓度变化；免疫组织化学和Westernblot法检测Ⅱ型胶原合成的变化。结果IGF组L-型钙通道最大电流密度为（5．447±O．208）pA／pF，显著高于对照组（4．925±0．316）pA／pF（P〈0．05）。激光共聚焦显微镜观察到IGF组钙离子荧光强度明显增强。免疫组织化学观察到IGF组Ⅱ型胶原染色明显比对照组更深。Western blot法行Ⅱ型胶原半定量分析，实验组22．96±4．92，显著高于对照组16．19±5．54（P〈0．05）。结论L型钙通道是IGF促进Ⅱ型胶原合成的一个重要途径。
Objective To investigate the effect of the L-type calcium channels on The type IT col- lagen synthesis of chondrocytes with patch-clamp. Methods The chondrocytes of rabbits, isolated and cultured in vitro, were divided into 2 groups： control group and insulin-like growth factor （IGF） group. Then, the L-type calcium channels was detected by patch-clamp, the calcium concentration by the laser confocal microscopy, and the type II collagen was quantitatively assessed by immunohistochemistrical stain and Western blotting. Results The L-type calcium channel current density was （5.447 ± 0. 208） pA/pF in IGF group, significantly higher than （4. 925 ± 0. 316 ） pA/pF in control group （ P 〈 0.05 ）. The fluo- rescence intensity of calcium in IGF group was significantly enhanced, and the type II collagen staining of immunohistochemical was significantly deeper than the control group. The semi-quantitative analysis of the type II collagen of the Western blot in IGF group was 22. 96 ±4. 92, and 16. 19 ±5.54 in the control group （P 〈 0.05 ）. Conclusion The L-type calcium channel is an important way of IGF to promote type II col- lagen synthesis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第7期1233-1235,共3页
Chinese Journal of Experimental Surgery
基金
江苏省自然科学基金资助项目（BK2011433）
江苏省苏北人民医院院级基金资助项目（NJPH200910）
关键词
钙通道
膜片钳
软骨细胞
Calcium channels
Patch clamp
Chondrocyte
