膜离子通道阻滞及细胞骨架破坏对拉伸应力刺激兔关节软骨细胞代谢的影响  

Effects of membrane ion channel block and cytoskeleton damage on tensile stress-stimulated metab- olism of articular cartilage cells

作　　者：乔梁段[1] 王平[1] 卫小春[1] 

机构地区：[1]山西医科大学第二医院骨科,太原030001

出　　处：《中华实验外科杂志》2013年第7期1364-1368,共5页Chinese Journal of Experimental Surgery

摘　　要：目的观察在拉伸应力作用下，细胞骨架（CSK）破坏剂和膜离子通道阻滞剂对体外培养兔膝关节软骨细胞代谢的影响。方法2月龄新西兰大白兔10只，无菌条件下剖取双膝，并将其消化为软骨细胞，并随机分为8组，即对照组，微管、微丝、中间纤维破坏组，Na＋、K＋、Ca2＋、Cl-通道阻滞组。在拉伸应力作用下干预3d后，苏木素-伊红（HE）染色观察其形态。采用酶联免疫吸附试验（ELASA）和阿尔新蓝染色检测各组标本上清液中基质金属蛋白酶（MMP）-13、金属蛋白酶组织抑制因子（TIMP）-1及糖胺聚糖（GAG）含量，采用逆转录一聚合酶链反应（RT—PCR）方法检测II型胶原（Col-II）、MMP-3、TIMP-1、微管蛋白、肌动蛋白、波形蛋白mRNA的相对表达量。结果HE染色软骨细胞形态由正常的软骨细胞变为排列不均匀，形态各异，细胞核深染，大小不一，细胞质减少。CSK破坏组上清液中GAG的浓度分别为15．6、18．0、14．0¨∥L，较对照组24．2肛∥L降低（P〈0．05）；微丝、中间纤维破坏组MMP-13的浓度分别为9．3、9．8μg/L，较对照组12．7μg/L降低（P〈0．05），中间纤维破坏组TIMP-1的浓度为5．1mg／L，较对照组2．3mg／L明显上升（P〈0．05）；Ca2＋、Cl-离子通道阻滞组GAG浓度分别为6．4、96．0μg/L，较对照组差异有统计学意义（P〈0．05）；K＋、Cl-通道阻滞组MMP-13浓度分别为8．8、6．7μg/L，较对照组均降低（P〈0．05）；各实验组软骨细胞Ⅱ型胶原mRNA相对表达量均下调（P〈0．05）；细胞骨架破坏的各组中，其相对应的蛋白mRNA表达量分别为54．4、52．1、51．3μg/L，较对照组减少且差异有统计学意义（P〈0．05）。结论在拉伸应力作用下，破坏CSK和阻滞离子通道使软骨细胞发生退化，其中微管、中间纤维和Na＋、Cl-通道可能与软骨细胞分泌的基质密切相关。Objective Discussion under tensile stress, Cytoskeleton （CSK） destruction agent and ion channel blockers on the metabolism of rabbit knee joint cartilage cells cultured in vitro. Methods 10 New Zealand white rabbits ,2-month-old ,under sterile conditions to taken knees, and digestion for cartilage cells, and randomly divided into eight groups, namely the control group, microtubules, microfilaments, inter- mediate filaments destruction group, Na＋ , K＋ , Ca2± , C1- channel blocking group. The tensile stress inter- vention 3 d,hematoxylin and eosin （HE） staining was observed morphology. Results HE staining mor- phology of chondroeytes from normal chondroeytes into are uniform, different patterns, with hyperchromatie nuclei, of different sizes, cytoplasmic reduced. The concentration of CSK in the supernatant of glycosamin- oglycan （GAG） failure group were 15.57,18.00,14.04 μg/L lower than the control group 24. 23μg/L P 〈 0.05 ） ; filaments, intermediate filaments were damaged in the concentration of matrix metalloproteinase （MMP） -13 was 9.28,9. 82μg/L decreased compared with the control group 12. 66μg/L（P 〈0. 05） ,in- termediate filaments were damaged in the concentration of tissue inhibitorof metalloproteinase （TIMP） -1 is 5.06 mg/L were increased significantly compared with the control group 2. 32 mg/L （ P 〈 0.05 ） ; Ca2 ± , C1- ion channel block GAG concentration was 6.40,95.95 μg/L compared with the control group, there was statistical significance ; K± , C1- channel block MMP-13 concentrations were 8.83,6. 67 μg/L were low- er than the control group （P 〈 01 05 ） ;the experimental group chondrocytes mRNA relative expression quan- tity was reduced （ P 〈 O. 05 ） ; Cytoskeleton damage in each group, its corresponding protein mRNA expres- sion quantity were 54.4,52. 1,51.3 ng/L1 than control group to reduce and statistical significance. Conclu- sion Under the action of tensile stress, damage CSK and block ion channel make cartilage cells occur degradation

关 键 词：软骨细胞 细胞骨架 离子通道 Ⅱ型胶原 糖胺多糖 

分 类 号：R441.1[医药卫生—诊断学]