Tsai Wen-Chung, Yu Tung-Yang, Lin Li-Ping, Lin Miao-Sui, Wu Yi-Cheng, Liao Chih-Hao, Pang Jong-Hwei S
a Department of Physical Medicine and Rehabilitation , Chang Gung Memorial Hospital at Linkou , Taoyuan City , Taiwan.
b College of Medicine , Chang Gung University , Taoyuan City , Taiwan.
Platelets. 2017 Jul;28(5):491-497. doi: 10.1080/09537104.2016.1227061. Epub 2016 Oct 26.
Platelet rich plasma (PRP) contains various cytokines and growth factors which may be beneficial to the healing process of injured muscle. The purpose of this study is to investigate the effect and molecular mechanism of PRP releasate on proliferation of skeletal muscle cells. Skeletal muscle cells intrinsic to Sprague-Dawley rats were treated with PRP releasate. Cell proliferation was evaluated by 3-[4,5-Dimethylthiazol- 2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and immunocytochemistry with Ki-67 stain. Flow cytometric analysis was used to evaluate the cell cycle progression. Western blot analysis was used to evaluate the protein expressions of PCNA, cyclin E1, cyclin A2, cyclin B1, cyclin dependent kinase (cdk)1 and cdk2. The results revealed that PRP releasate enhanced proliferation of skeletal muscle cells by shifting cells from G1 phase to S phase and G2/M phases. Ki-67 stain revealed the increase of proliferative capability after PRP releasate treatment. Protein expressions including cyclin A2, cyclin B1, cdk1, cdk2 and PCNA were up-regulated by PRP releasate in a dose-dependent manner. It was concluded that PRP releasate promoted proliferation of skeletal muscle cells in association with the up-regulated protein expressions of PCNA, cyclin A2, cyclin B1, cdk1 and cdk2.
富含血小板血浆(PRP)含有多种细胞因子和生长因子,这些因子可能对受损肌肉的愈合过程有益。本研究的目的是探讨PRP释放物对骨骼肌细胞增殖的影响及其分子机制。用PRP释放物处理Sprague-Dawley大鼠的原代骨骼肌细胞。通过3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐(MTT)法和Ki-67染色免疫细胞化学法评估细胞增殖。采用流式细胞术分析评估细胞周期进程。采用蛋白质印迹法分析评估增殖细胞核抗原(PCNA)、细胞周期蛋白E1、细胞周期蛋白A2、细胞周期蛋白B1、细胞周期蛋白依赖性激酶(cdk)1和cdk2的蛋白表达。结果显示,PRP释放物通过将细胞从G1期转移至S期和G2/M期来增强骨骼肌细胞的增殖。Ki-67染色显示PRP释放物处理后增殖能力增强。PRP释放物以剂量依赖性方式上调细胞周期蛋白A2、细胞周期蛋白B1、cdk1、cdk2和PCNA的蛋白表达。结论是,PRP释放物通过上调PCNA、细胞周期蛋白A2、细胞周期蛋白B1、cdk1和cdk2的蛋白表达促进骨骼肌细胞增殖。