外敷温经药酒联合针刺对骨质疏松大鼠的影响

The effect of external application of warm meridian medicinal wine combined with acupuncture on osteoporosis rats

目的探究外敷温经药酒联合针刺对骨质疏松大鼠的影响。方法选取SPF级雌性大鼠40只,分为正常组、模型组、针刺组、联合组,每组10只。除正常组外其余均建立骨质疏松(osteoporosis,OP)大鼠模型。建模成功后正常组、模型组不予处理,针刺组入穴关元、三阴交(双)、肾俞(双)、后三里(双)后连接电针仪治疗,5 d/周,每日一次,20 min/次,共12周。联合组在针刺治疗基础上外敷温经药酒,每日两次,共12周。用ELISA法检测血清中骨钙素(OC)、吡啶酚(PYD)、Ⅰ型前胶原羧基端肽(PICP)水平;用计算机断层扫描检测骨小梁厚度(Tb.Th)、骨小梁间隔(Tb.Sp)、骨密度(bone mineral density,BMD)、皮质骨面积(Ct.Ar);运用TUNEL法检测成骨细胞凋亡;通过HE染色观察股骨组织病理改变;通过Western blot法测Notch1、Notch2、Jagged1、Jagged2。结果与正常组比较,模型组PYD、Tb.Sp、成骨细胞凋亡率升高,OC、PICP、Tb.Th、BMD、Ct.A、Notch1、Notch2、Jagged1、Jagged2降低(P<0.05);与模型组比较,针刺组PYD、Tb.Sp、成骨细胞凋亡率降低,OC、PICP、Tb.Th、BMD、Ct.A、Notch1、Notch2、Jagged1、Jagged2升高(P<0.05);与针刺组比较,联合组PYD、Tb.Sp、成骨细胞凋亡率降低,OC、PICP、Tb.Th、BMD、Ct.A、Notch1、Notch2、Jagged1、Jagged2升高(P<0.05);正常组大鼠骨形态结构良好;模型组骨损伤严重;针刺组与联合组骨形态结构有所改善,其中联合组改善更为良好。结论外敷温经药酒联合针刺对骨质疏松大鼠的骨钙素、成骨细胞活性及Notch信号通路等的改善作用良好,建议推广使用。

Objective Exploring the Effect of External Application of Warm Meridian Medicinal Wine Combined with Acupuncture on Osteoporosis Rats.Methods Forty SPF grade female rats were selected and divided into normal group,model group,acupuncture group,and combination group,with 10 rats per group.Except for the normal group,osteoporosis(OP)models were established in all other groups.After successful modeling,the normal group and model group were not treated.The acupuncture group was treated with acupoints at Guanyuan,Sanyinjiao(Shuang),Shenshu(Shuang),and Housanli(Shuang),and then connected to an electroacupuncture instrument for 5 days per week,20 minutes per day,for a total of 12 weeks.On the basis of acupuncture treatment,the combination group applied warm meridian medicinal wine twice a day for a total of 12 weeks.The serum levels of osteocalcin(OC),pyridinol(PYD),and type I procollagen carboxy terminal peptide(PICP)were detected using ELISA.The bone trabecular thickness(Tb.Th),trabecular septa(Tb.Sp),bone density(BMD),and cortical bone area(Ct.Ar)were measured using computed tomography.Osteoblast apoptosis was detected using TUNEL,and pathological changes in femoral tissue were observed using HE staining.Notch1,Notch2,Jagged1,and Jagged2 were measured using Western blot.Results Compared with the normal group,the model group PYD and Tb Sp,increased apoptosis rate of osteoblasts,OC,PICP,Tb Th,BMD,Ct A.Notch1,Notch2,Jagged1,and Jagged2 decreased(P<0.05);Compared with the model group,the acupuncture group had PYD and Tb Sp,decreased apoptosis rate of osteoblasts,OC,PICP,Tb Th,BMD,Ct A.Notch1,Notch2,Jagged1,and Jagged2 increased(P<0.05);Compared with the acupuncture group,the combination group had PYD and Tb Sp,decreased apoptosis rate of osteoblasts,OC,PICP,Tb Th,BMD,Ct A.Notch1,Notch2,Jagged1,and Jagged2 increased(P<0.05);The bone morphology and structure of the normal group rats are good;The model group had severe bone damage;The bone morphology and structure of the acupuncture group and the combination group have improved,with the combination group showing better improvement.Conclusion The combination of external application of Wen Jing Yao Jiu and acupuncture has a good improvement effect on osteocalcin,osteoblast activity,and Notch signaling pathway in osteoporosis rats,andit is recommendedto promote its use.