实用医学杂志 ›› 2022, Vol. 38 ›› Issue (23): 2947-2951.doi: 10.3969/j.issn.1006⁃5725.2022.23.010

• 临床研究 • 上一篇    下一篇

重组人血小板生成素对凝血酶诱导血小板凋亡的调节作用 

周先桃1 李瑜1 沈阳2 王正华2    

  1. 1 武汉血液中心(武汉 430030);2 湖北省中西医结合医院(武汉 430015)

  • 出版日期:2022-12-10 发布日期:2022-12-10
  • 基金资助:
    湖北省卫生健康委科研基金项目(编号:2041ZF315)

Regulatory effect of recombinant human thrombopoietin on the cell apoptosis of platelet induced by throm⁃ bin

ZHOU Xiantao*,LI Yu,SHEN Yang,WANG Zhenghua.   

  1. Wuhan Blood Center,Wuhan 430030,China 

  • Online:2022-12-10 Published:2022-12-10

摘要:

目的 探讨重组人血小板生成素(rhTPO)对凝血酶(TMB)诱导的血小板凋亡的调节机制。 方法 常规制备洗涤血小板,分组如下:Normal 组、TMB 组、rhTPO(低、中、高)剂量组。检测各组血小板 中线粒体的完整性、细胞凋亡率以及 ROS 的含量,检测 Opa1、Bcl⁃xl、Mfn2、Fis⁃1、Drp⁃1、Cyt⁃c 和 Bak 的表 达。结果 与 Normal 组相比,TMD 组、rhTPO(低、中、高)组中血小板中完整线粒体的比例及 Opa1、Bcl⁃xl、 Mfn2 的 mRNA 和蛋白的表达明显降低,细胞的凋亡率和 ROS 的含量及 Fis⁃1、Drp⁃1、Cyt⁃c 和 Bak 的 mRNA 和蛋白的表达明显升高,rhTPO(低、中、高)剂量组能明显逆转这些趋势,并且具有明显的剂量依赖性,差 异均具有统计意义(< 0.05)。结论 rhTPO 能明显抑制 TMD 诱导的血小板的细胞凋亡,这可能与抑制 ROS 的富集,改善线粒体功能有关。

关键词:

重组人血小板生成素, 血小板, 细胞凋亡, 凝血酶, 线粒体

Abstract:

Objective To investigate the regulatory mechanism of recombinant human thrombopoietin (rhTPO)on the cell apoptosis of platelet induced by thrombin(TMB). Methods Washed platelets were routinely prepared and divided into five groups:normal control,TMB group and three rhTPO groups with low,medium and high dose respectively. MitoTracker Red CMXRos staining was used to detect the integrity of mitochondria,the apoptosis rate,the content of reactive oxygen species(ROS). The expression levels of Opa1,Bcl⁃xl,Mfn2,Fis⁃1, Drp⁃1,Cyt⁃c and Bak were also detected. Results Compared with the normal group,the proportion of intact mito⁃ chondria in platelets,the mRNA and protein expressions of Opa1,Bcl ⁃xl and Mfn2,in the TMD group and the rhTPO low,medium and high groups,were significantly decreased. The apoptosis rate and the content of ROS and the mRNA and protein expressions of Fis⁃1,Drp⁃1,Cyt⁃c and Bak were significantly increased. The rhTPO(low, medium,and high)groups could obviously reverse these trends,and had a significant dose ⁃ dependent manner. The differences were statistically significant(all < 0.05). Conclusion rhTPO can significantly inhibit the cell apoptosis of platelet induced by TMD,which may be related to inhibiting ROS accumulation and improving mito⁃ chondrial function.

Key words:

recombinant human thrombopoietin, platelet, apoptosis, thrombin, mitochondrion