摘要
目的 探讨电针联合阿仑膦酸钠对去卵巢大鼠骨质疏松症的改善作用及对 Integrinα2/FAK/Runx2 通路调节作用.方法 选取30 只大鼠随机分为假手术组(6 只)及造模组(24 只),采用手术切除卵巢法建立骨质疏松症大鼠模型.将造模后的大鼠随机分为骨质疏松模型组、电针治疗组、阿仑膦酸钠组及联合治疗组,每组 6 只.电针治疗组、阿仑膦酸钠组及联合治疗组分别按照相应干预方法干预 8 周.通过酶联免疫吸附(ELISA)试剂盒测定大鼠血清碱性磷酸酶(ALP)、骨钙素(BGP)、抗酒石酸酸性磷酸酶(TRACP)-5b、Ⅰ型胶原羧基末端肽(ICIP)、I型胶原氨基端延长肽(PINP)水平;双能X射线测定大鼠股骨骨密度及骨矿物含量;骨生物力学测定仪测定大鼠骨生物力学指标;苏木精-伊红(HE)染色法检查股骨组织病理学变化;实时定量聚合酶链式反应(RT-qPCR)测定股骨组织Integrinα2、FAK及Runx2 mRNA水平;免疫印记法(Western blot)测定股骨组织Integrinα2、FAK及Runx2 蛋白水平.结果 与假手术组比较,骨质疏松模型组血清ALP、BGP、TRACP-5b、ICIP、PINP水平显著升高(P＜0.05),骨密度、骨矿物含量、最大载荷、最大应力、刚度,股骨组织Integrinα2、FAK及Runx2 mRNA及蛋白水平显著降低(P＜0.05);与骨质疏松模型组比较,电针治疗组、阿仑膦酸钠组及联合治疗组血清ALP、BGP、TRACP-5b、ICIP、PINP水平显著降低(P＜0.05),骨密度、骨矿物含量、最大载荷、最大应力、刚度,股骨组织Integrinα2、FAK及Runx2 mRNA及蛋白水平显著升高(P＜0.05);与电针治疗组及阿仑膦酸钠组比较,联合治疗组血清ALP、BGP、TRACP-5b、ICIP、PINP水平显著降低(P＜0.05),骨密度、骨矿物含量、最大载荷、最大应力、刚度,股骨组织Integrinα2、FAK及Runx2 mRNA及蛋白水平显著升高(P＜0.05).结论 电针联合阿仑膦酸钠能够显著提高绝经后骨质疏松症大鼠骨密度,抑制骨质疏松病理进展,改善大鼠骨代谢及骨生物力学改变,其机制可能与调节Integrinα2/FAK/Runx2 通路有关.
Abstract
Objective To study the effect of electroacupuncture combined with alendronate sodium on the improvement of osteoporosis in ovariectomized rats and the regulation of integrin-α2/FAK/Runx2 pathway.Methods Thirty rats were randomly divided into sham operation group(6 rats)and modeling group(24 rats).The postmenopausal osteoporosis rat model was established by surgical ovariectomy of the rats in the modeling group.The rats after modeling were randomly divided into osteoporosis model group,electroacupuncture group,alendronate sodium group,and combined treatment group,with 6 rats per group.Electroacupuncture stimulated Shenshu and Zusanli acupoints of rats.The intervention continued for 8 weeks.Rats in alendronate group received alendronate sodium by intragastric administration of 0.5 mg/kg once a day for 8 weeks.In the combined treatment group,rats received alendronate sodium and electroacupuncture therapy for 8 weeks.Serum levels of alkaline phosphatase(ALP),osteocalcin(BGP),tartrate-resistant acid phosphatase(TRACP)-5b,carboxyl terminal peptide of type Ⅰ collagen(ICIP),and amino terminal prolongating peptide of type Ⅰ collagen(PINP)were determined with enzyme-linked immunosorbent assay(ELISA)kit.Bone mineral content and bone mineral density of rat femurs were determined with dual-energy X-ray bone densitometer and bone mechanical property tester.Histopathological changes of rat femurs were examined with hematoxylin-eosin(HE)staining.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to determine the mRNA levels of integrin-α2,FAK,and Runx2 in the femur.Immunoblot was used to determine the protein levels of integrin-α2,FAK,and Runx2 in the femur.Results Compared to those in the normal control group,serum levels of ALP,BGP,TRACP-5b,ICIP,and PINP in osteoporosis model group increased significantly(P＜0.05),and bone mineral content,maximum load,maximum stress,and stiffness decreased significantly(P＜0.05).The mRNA and protein levels of integrinα2,FAK,and Runx2 in femur tissue decreased significantly(P＜0.05).Compared to those in the osteoporosis model group,serum levels of ALP,BGP,TRACP-5b,ICIP,and PINP in electroacupuncture group,alendronate sodium group,and combined treatment group decreased significantly(P＜0.05),and bone mineral density,bone mineral content,maximum load,maximum stress,and stiffness increased significantly(P＜0.05).The mRNA and protein levels of integrinα2,FAK,and Runx2 in the femur increased significantly(P＜0.05).Compared to those in the electroacupuncture treatment group and alendronate sodium group,serum ALP,BGP,TRACP-5b,ICIP,and PINP levels in combined treatment group decreased significantly(P＜0.05),and bone mineral density,bone mineral content,maximum load,maximum stress,and stiffness increased significantly(P＜0.05).The mRNA and protein levels of integrinα2,FAK,and Runx2 in the femur increased significantly(P＜0.05).Conclusion Electroacupuncture combined with alendronate sodium significantly increases bone mineral density in postmenopausal rats with osteoporosis,inhibits the pathological progression of osteoporosis,improve bone metabolism and bone biomechanical changes in rats.The mechanism may be related to the regulation of integrin-α2/FAK/Runx2 pathway.
维普
万方数据