摘要
目的 探究汉黄芩素(WOG)调节腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)信号通路对骨质疏松(osteoporosis,OP)大鼠骨折愈合的影响。方法 将60只SPF级SD雌性大鼠随机分为假手术组(Sham组)、骨质疏松性骨折(OPF)模型组(Model组)、低剂量WOG组(WOG-L组,50mg/kg)、高剂量WOG组(WOG-H组,100 mg/kg)和高剂量WOG+AMPK抑制剂Compound C组(WOG-H+CC组,100 mg/kg WOG+0.2 mg/kg Compound C),每组12只。Model组、WOG-L组、WOG-H组、WOG-H+CC组采用双侧卵巢切除术与右侧股骨干骨折髓内固定术构建骨质疏松性骨折(OPF)大鼠模型。放射性检查评估骨折愈合情况。采用双能X线骨密度扫描仪检测大鼠股骨痂骨密度(BMD)。通过三点弯曲实验和压缩实验对大鼠股骨骨生物力学进行检测。比色法检测大鼠血清骨钙素(OC)、I型前胶原羧基端肽(PICP)水平。HE染色观察骨折处骨痂的病理学变化。Western blot法检测股骨组织中AMPK/SIRT1信号通路相关蛋白表达。结果 与Sham组相比,Model组大鼠骨折愈合评分、股骨上、下端骨痂BMD、骨痂最大位移、最大应力和最大负荷、血清OC、PICP水平、股骨组织中AMPK磷酸化水平、SIRT1蛋白表达显著降低(P <0.05),骨折端未见骨痂生长,骨折线清晰,骨小梁较细,数量减少,间隙增大,结构紊乱疏松。与Model组相比,WOG-H组大鼠骨折愈合评分、股骨上、下端骨痂BMD、骨痂最大位移、最大应力和最大负荷、血清OC、PICP水平、股骨组织中AMPK磷酸化水平、SIRT1蛋白表达显著升高(P <0.05),有连续性骨痂包绕贯穿于骨折端,髓腔再通,骨折线模糊不清,骨小梁形态较完整。Compound C减弱了WOG对OPF大鼠骨折愈合的促进作用。结论 WOG可能通过激活AMPK/SIRT1信号通路促进OPF大鼠的骨折愈合。
Objective To investigate the effect of wogonin(WOG)on bone fracture healing in rats with osteoporosis(OP)by regulating AMP-activated protein kinase(AMPK)/silent information regulator 1(SIRT1)signaling pathway.Methods Sixty female SPF SD rats were randomly divided into sham operation group(Sham group),osteoporotic fracture(OPF)model group(Model group),low-dose WOG group(WOG-L group,50 mg/kg),high-dose WOG group(WOG-H group,100 mg/kg)and high-dose WOG+AMPK inhibitor Compound C group(WOG-H+CC group,100 mg/kg WOG+0.2 mg/kg Compound C),with 12 rats in each group.Model group,WOG-L group,WOG-H group and WOG-H+CC group were selected to establish the rat model of osteoporotic fracture(OPF)by bilateral oophorectomy and intramedullary fixation of right femoral shaft fracture.Radiological examination was performed to evaluate the fracture healing.Bone mineral density(BMD)of rat femoral callus was measured by dual energy X-ray absorptiometry.The biomechanics of rat femoral bone was tested by three-point bending test and compression test.Serum osteocalcin(OC)and procollagen typeⅠcarboxy-terminal peptide(PICP)were measured by colorimetry.HE staining was used to observe the pathological changes of callus at the fracture site.Western blot was used to detect the expression of AMPK/SIRT1 signal pathway related proteins in femur.Results Compared with the Sham group,the fracture healing score,BMD of upper and lower femur callus,the maximum displacement,maximum stress and maximum load of callus,the serum OC and PICP levels,AMPK phosphorylation level in femur tissue and SIRT1 protein expression in Model group were obviously reduced(P<0.05),no callus growth was found at the fracture end,the fracture line was clear,the bone trabecula was thin,the number was reduced,the gap was increased,and the structure was disordered and loose.Compared with the Model group,the fracture healing score,BMD of upper and lower femur callus,the maximum displacement,maximum stress and maximum load of callus,the serum OC and PICP levels,AMPK phosphorylation level in femur tissue and SIRT1 protein expression in the WOG-H group were obviously increased(P<0.05),there was continuous callus wrapping and penetrating the fracture end,the medullary cavity was reopened,the fracture line was unclear,and the shape of bone trabecula was relatively complete.Compound C weakened the promoting effect of WOG on fracture healing in OPF rats.Conclusion WOG may promote fracture healing in OPF rats by activating AMPK/SIRT1signaling pathway.
作者
李庆贵
陈康
郭召
骞立刚
LI Qinggui;CHEN Kang;GUO Zhao;QIAN Ligang(Affiliated Hospital of Hebei University,Baoding 071000,China)
出处
《实用医学杂志》
CAS
北大核心
2023年第4期410-416,共7页
The Journal of Practical Medicine
基金
河北省自然科学基金项目(编号:H2021201055)。
