虫草素通过抑制氧化应激和炎症反应对骨折愈合的影响研究

Study on the effect of cordycepin on fracture healing by inhibiting oxidative stress and inflammation

关键词:  虫草素  骨折  氧化应激  炎症反应

Key words:  cordycepin  fracture  oxidative stress  inflammation

摘要:

[目的] 探讨虫草素治疗骨折愈合的作用及机制。[方法] 50只SPF级SD大鼠（240～270 g），采用标准的大鼠闭合性股骨横行骨折内固定模型，并腹腔注射给予虫草素治疗；采用微型CT扫描仪（Micro-CT）对骨折大鼠股骨进行放射学检测，并用骨体积分数（BV/TV），骨小梁厚度（Tb.Th），骨小梁数（Tb.N）和骨密度（BMD）定量分析；采用商用融合检测试剂盒检测组织中骨源碱性磷酸酶（BAP）、核因子κB受体活化因子配体（RANKL）和抗酒石酸酸性磷酸酶（TRAP）的含量；采用苏木精-伊红染色（HE）、番红固绿染色和马松染色检测骨折大鼠股骨的病理学变化；采用酶联免疫吸附试验（ELISA）试剂盒检测骨折大鼠血清中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）的含量；采用商用融合检测试剂盒检测骨折大鼠血清丙二醛（MDA）含量及组织中MDA和过氧化氢酶（CAT）的含量。[结果] 与模型组相比，给予虫草素治疗大鼠的骨折部位明显愈合，并且Tb.Th，BV/TV，BMD，和Tb.N显著增加（P<0.05）；给予虫草素治疗大鼠的股骨组织中骨源碱性磷酸酶（BAP）、RANKL和抗酒石酸酸性磷酸酶（TRAP）含量呈剂量依赖性显著减少（P<0.05）；HE染色结果表明虫草素5 mg/kg和10 mg/kg组大鼠观察板层状骨排列整齐，编织骨减少；番红固绿染色结果表明虫草素5 mg/kg和10 mg/kg组大鼠观察到软骨组织减少，成骨组织增加；马松染色结果表明虫草素5 mg/kg和10 mg/kg组观察到成熟骨组织；虫草素呈剂量依赖性显著减小骨折大鼠IL-1β、IL-6和TNF-α的含量（P<0.05）；同时，虫草素呈剂量依赖性显著减小骨折大鼠血清MDA含量及组织中MDA和CAT的含量（P<0.05）。[结论] 虫草素对骨折愈合具有潜在的促进作用，并且这种作用可能与抗氧化应激和炎症反应有关。

Abstract:

[Objective] To explore the effect and mechanism of cordycepin in the treatment of fracture healing. [Methods] 50 SPF SD rats were selected to established fracture healing model,which were treated with cordycepin. The femurs of fracture rats were examined by Micro-CT and quantitatively analyzed by Tb.Th,BV/TV,BMDand Tb.N,the contents of bone alkaline phosphatase(BAP),RANKL and tartrate-resistant acid phosphatase(TRAP) in tissues were detected by commercial kit. The contents of bone alkaline phosphatase(ALP),bone alkaline phosphatase(RANKL) and tartrate-resistant acid phosphatase(TRAP) in tissues were detected by commercial kit. The pathological changes of femur of fract'ure rats were detected by hematoxylin-eosin staining,safranin O staining and Masson staining. The contents of interleukin-1β,interleukin-6 and tumor necrosis factor-α in serum of fracture rats were detected by ELISA kit. The contents of MDA in serum and MDA and CAT in tissue of fracture rats were detected by commercial kit. [Results] Compared with the model group,the fracture site of rats treated with cordycepin healed obviously,and Tb.Th,BV/TV,BMD and Tb.N increased significantly(P<0.05),and cordycepin significantly decreased the contents of bone alkaline phosphatase (BAP),RANKL and tartrate resistant acid phosphatase (TRAP) in femoral tissue of fracture rats in a dose-dependent manner. The HE staining results showed that the cordycepin 5mg/kg and 10mg/kg group rats observed lamellar bones arranged neatly and the woven bones were reduced;the safranine fast green staining results showed that the cordycepin 5 mg/kg and 10 mg/kg groups had cartilage. The tissue decreased and the osteogenic tissue increased;Masson staining results showed that mature bone tissue was observed in cordycepin 5 mg/kg and 10 mg/kg groups. Cordycepin significantly decreased the contents of IL-1β,IL-6 and TNF-α in fracture rats in a dose-dependent manner,and significantly decreased the contents of MDA in serum and MDA and CAT in tissues of fracture rats in a dose-dependent manner. [Conclusion] Cordycepin has a potential promoting effect on fracture healing,and this effect may be related to antioxidant stress and inflammation.