摘要
目的探讨Apelin-13调控VEGF/PI3K/Akt信号通路对大鼠股骨骨折愈合的影响。方法将40只雄性SD大鼠随机分成假手术组(sham组)、模型组(Model组)、Apelin-13低剂量组[Apelin-13 L组,25μg/(kg·d)]和Apelin-13高剂量组[Apelin-13 H组,75μg/(kg·d)],每组10只。建立大鼠股骨骨折模型,立即给予相应药物处理,治疗4周。采用Micro CT和X射线分别检测各组大鼠股骨显微参数和骨痂直径;苏木精-伊红(HE)染色观察骨组织病理变化;酶联免疫吸附测定(ELISA)检测血清中成骨标记物骨源性碱性磷酸酶(BALP)、Ⅰ型前胶原氨基端前肽(PINP)和破骨标记物抗酒石酸酸性磷酸酶-5b(TRACP-5b)、Ⅰ型胶原C端肽(CTX)表达水平;qRT-PCR检测骨痂组织中血管内皮生长因子A(VEGFA)mRNA表达水平;Western blot检测骨痂组织中VEGF/PI3K/Akt信号通路相关蛋白表达水平。结果与sham组比较,Model组大鼠股骨病理损伤较严重,股骨骨密度、组织矿物质密度、骨体积分数和X射线评分均降低,血清中BALP和PINP水平降低,而TRACP-5b、CTX含量均增加,骨痂组织中VEGFA mRNA以及VEGFA、VEGFR2、PI3K p85、p-Akt等蛋白表达水平降低,差异有统计学意义(P<0.05)。与Model组比较,Apelin-13 H组大鼠股骨病理损伤改善明显,股骨骨密度、组织矿物质密度、骨体积分数、X射线评分和骨痂直径均增加,血清中BALP和PINP含量增加,TRACP-5b和CTX含量减少,骨痂组织中VEGFA mRNA以及VEGFA、VEGFR2、PI3K p85、p-Akt蛋白表达水平增加,差异有统计学意义(P<0.05),而Apelin-13 L组与Model组比较,相关指标差异无统计学意义(P>0.05)。结论外源性Apelin-13可促进大鼠股骨骨折愈合,其机制可能与VEGF/PI3K/Akt信号通路的激活有关。
Objective To investigate the effect of Apelin-13 regulation of VEGF/PI3 K/Akt signaling on femoral fracture healing.Methods Forty male SD rats were randomly divided into sham group, Model group, Apelin-13 low-dose group[Apelin-13 L, 25 μg/(kg·d)] and Apelin-13 high-dose group[apelin-13 H, 75 μg/(kg·d)], and 10 rats in each group. The model of femoral fracture was established and immediately treated with appropriate drugs for 4 weeks. Micro CT and X-ray were used to detect bone microscopic parameters and callus diameter. HE staining was used to observe bone tissue pathological changes. ELISA was used to detect the expression levels of serum bone markers BALP, PINP and osteoclast TRACP-5 b, CTX. qRT-PCR was used to detect VEGFA mRNA expression levels in callus tissues, and Western blot was used to detect the expression levels of related proteins in the VEGF/PI3 K/Akt signaling pathway in callus.Results Compared with the sham group, the Model group had severe pathological damage of femur and decreased femur bone density, tissue mineral density, bone mass fraction and X-ray score. BALP and PINP levels in serum decreased, while TRACP-5 b and CTX levels increased, and VEGFA mRNA, VEGFA, VEGFR2, PI3 K p85 and p-Akt expression levels in callus decreased. Compared with the Model group, the pathological injury of the femur in the Apelin-13 H group improved. Bone mineral density, tissue mineral density, bone volume fraction, X-ray score and callus diameter all increased. BALP and PINP levels in serum increased(P<0.05), while TRACP-5 b and CTX levels decreased, and VEGFA mRNA, VEGFA, VEGFR2, PI3 K p85 and p-Akt expression levels in callus increased. There was no significant difference between Apelin-13 L group and Model group(P>0.05).Conclusion Exogenous Apelin-13 can promote femoral fracture healing in rats, and its mechanism may be related to the activation of the VEGF/PI3 K/Akt signaling pathway.
作者
曾昭池
汤勇
张先慧
朱志勇
吴开元
袁永端
Zeng Zhaochi;Tang Yong;Zhang Xianhui(Dept of Orthopaedics,The 922nd Hospital of The Joint Service Support Force of The PLA,Hengyang 421002)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第1期48-53,共6页
Acta Universitatis Medicinalis Anhui
基金
湖南省卫健委科研计划项目(编号:C2015-29)。
