诱导型一氧化氮合酶对骨骼肌损伤再生的影响

Role of iNOS during wound healing of skeletal muscle

目的观察骨骼肌损伤再生中的细胞形态学变化和损伤后诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达水平;观察巨噬细胞上清液对iNOS表达水平和骨骼肌再生的影响。方法制备SD大鼠骨骼肌损伤模型,损伤3 d后获取骨骼肌,进行HE染色、iNOS免疫组化分析;利用原代培养的巨噬细胞上清液处理小鼠C2C12成肌细胞株,RT-PCR技术探讨巨噬细胞和成肌细胞相互作用的分子机制。结果 HE染色可见,骨骼肌损伤处有新的血管产生并进入损伤的骨骼肌内;损伤部位iNOS表达强阳性,生发区iNOS表达呈阳性,正常与损伤细胞表达差异明显;巨噬细胞上清液处理小鼠C2C12成肌细胞株能够促进iNOS和成肌细胞因子Myo D的表达。结论骨骼肌损伤过程中,iNOS的表达增加能够促进骨骼肌的再生,加快骨骼肌的损伤修复。

Objective To observe the morphological changes and iNOS expression at cellular damaged zone during the wound healing of denervated muscle;To determine the affection of the expression of iNOS induced by supernatant of macrophages. Methods The damaged denervated muscle, three days after the damaged muscle model of rat was established, was harvested for H.E and immunohistochemical staining of iNOS. The cultured C2C12 cells were treated with supernatant of macrophages. The treated C2C12 cells were analyzed using RT-PCR method. Results New capillaries near the damaged muscle and high expression of iNOS could be observed. The expression of iNOS was different between the damaged and undamaged zones in muscle cells. The supernatant of macrophages could induce the expression of iNOS and MyoD genes. Conclusion The high expression of iNOS can accelerate the regeneration of denervated muscle during wound healing.