[1]朱志恒,吴希晨,丁佳滢,等.凝血酶促大鼠软骨细胞生长、形态转变和抑制胶原合成的作用[J].中国中医骨伤科杂志,2022,30(11):1-6.
　ZHU Zhiheng,WU Xichen,DING Jiaying,et al.Efficacy of Thrombin on the Proliferation Morphological Transformation and Collagen Synthesis of Rat Chondrocytes[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2022,30(11):1-6.
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凝血酶促大鼠软骨细胞生长、形态转变和抑制胶原合成的作用()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第30卷
期数:
2022年11期
页码:
1-6
栏目:
实验研究
出版日期:
2022-11-15

文章信息/Info

Title:
Efficacy of Thrombin on the Proliferation Morphological Transformation and Collagen Synthesis of Rat Chondrocytes
文章编号:
1005-0205(2022)11-0001-06
作者:
朱志恒1; 2; 吴希晨1; 2; 丁佳滢1; 2; 王学宗3; 曹月龙3; 郑昱新3; 丁道芳1; 2△
1上海中医药大学康复医学院(上海,201203) 2上海中医药大学康复医学研究所 3上海中医药大学附属曙光医院石氏伤科医学中心
Author(s):
ZHU Zhiheng1; 2; WU Xichen1; 2; DING Jiaying1; 2; WANG Xuezong3; CAO Yuelong3; ZHENG Yuxin3; DING Daofang1; 2△
1School of Rehabilitation Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 2Institute of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 3Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
关键词:
凝血酶;  软骨细胞;  胶原合成;  增殖;  形态
Keywords:
thrombin;  chondrocyte;  collagen synthesis;  proliferation;  morphology
分类号:
R-33
文献标志码:
A
摘要:
目的:观察凝血酶对大鼠软骨细胞生长、形态和胶原合成的影响。方法:提取新生SD大鼠软骨细胞,取P1代细胞进行实验,设对照组和凝血酶组,凝血酶组加入20 U/mL凝血酶进行干预。倒置显微镜下观察两组细胞形态,并使用Edu试剂盒检测细胞增殖; 实时荧光定量聚合酶链式反应(PCR)法检测软骨细胞外基质成分Col2α1、Acan、Eln、Comp,炎症因子IL-6、TNF-α,炎症趋化因子Ccl2、Ccl7、Cxcl6、Cxcl16和胶原合成相关酶P3H1、Colgalt1、Lox的表达; 免疫细胞化学检测Lox的表达; 免疫荧光检测Col2α1的表达; Western Blot法检测分解代谢基因MMP9、MMP13、COX2的表达。结果:与对照组相比,凝血酶组的软骨细胞多为长梭形或不规则形状,且增殖明显; 凝血酶组Col2α1、Acan、Eln、Comp、P3H1、Colgalt1、Lox的mRNA表达下调(P<0.05),IL-6、TNF-α、Ccl2、Ccl7、Cxcl6、Cxcl16 mRNA表达上调(P<0.05),Lox和 Col2α1表达下调(P<0.05),MMP9、MMP13、COX2蛋白表达上调(P<0.05)。结论:凝血酶能够促进大鼠软骨细胞增殖和纤维样形态转变,提高炎症因子和分解代谢基因的表达,降低胶原合成相关酶表达而抑制胶原合成。
Abstract:
Objective:To observe the efficacy of thrombin on the growth, morphology and collagen synthesis of rat chondrocytes.Methods:Primary chondrocytes of neonatal SD rats were isolated, and all experiments were performed with cells from passage one.Chondrocytes were divided into two groups: control group and thrombin group.Chondrocytes were cultured with 20 U/mL thrombin in thrombin group.The cell morphology of the two groups was observed under an inverted microscope, and the cell proliferation was detected by an Edu kit.qRT-PCR was used to detect the expression of extracellular matrix(Col2α1, Acan, Eln, Comp), inflammatory factors(IL-6, TNF-α), chemokines(Ccl2, Ccl7, Cxcl6, Cxcl16)and enzymes that were involved in collagen synthesis(P3H1, Colgalt1, Lox).The expression of Lox was detected by immunocytochemistry.The expression of Col2α1 was detected by immunofluorescence.The expressions of catabolic genes MMP9, MMP13 and COX2 were detected by Western Blot.Results:Compared with the control group, the morphology of chondrocytes in thrombin group became long spindle, and the proliferative efficacy of thrombin on chondrocytes was significant.The mRNA levels of Col2α1, Acan, Eln, Comp, P3H1, Colgalt1, Lox were down-regulated in thrombin group(P<0.05), and the expressions of IL-6,TNF-α,Ccl2,Ccl7,Cxcl6,Cxcl16 mRNA were up-regulated(P<0.05).The expression of Lox and Col2α1 of thrombin group were increased.Compared with the control group, the protein levels of MMP9, MMP13 and COX2 protein in thrombin group were increased(P<0.05).Conclusion:Thrombin can promote the proliferation of rat chondrocytes and the morphology transformation towards fibroblastic chondrocytes, and inhibit collagen synthesis by increasing the expression of inflammatory factors and catabolic genes and reducing the activity of collagen synthesis related enzymes.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(81902306,82174406) △通信作者 E-mail:dingdaofang@shutcm.edu.cn
更新日期/Last Update: 2022-11-10