电针康复干预Wnt/β-catenin通路对骨质疏松大鼠的影响

The effect of electroacupuncture rehabilitation intervention on Wnt/β-catenin pathway in osteoporotic rats

目的探究电针康复干预Wnt/β-catenin通路对骨质疏松(osteoporosis,OP)模型大鼠的治疗作用及影响机制。方法随机将48只雌性SD大鼠分为空白组、模型组、实验组和联合组(针刺+Wnt拮抗剂),每组12只。空白组不作处理,其他组去卵巢法建立OP模型大鼠。实验组给予针刺治疗,联合组在针刺治疗的同时采用Wnt拮抗剂DKK1皮下注射干预,其他两组常规饲料喂养,连续干预6周。对比各组大鼠骨组织形态学参数变化;运用ELISA法检测血清TNF-α、IL-6、COX-2、MMP-3变化;通过TUNEL检测试剂盒分析大鼠骨组织细胞凋亡;运用免疫组化法检测GPX4蛋白表达;运用Western blot及RT-PCR检测骨组织中β-catenin、Wnt3a蛋白表达及mRNA表达水平。结果与空白组比较,模型组大鼠骨组织形态出现明显改变,TNF-α、IL-6、COX-2、MMP-3、Wnt3a、β-catenin蛋白及mRNA水平均升高(P值均<0.05),GPX4表达降低(P<0.05),病理损伤严重;与模型组对比,实验组大鼠骨组织形态学发生显著改善,TNF-α、IL-6、COX-2、MMP-3、Wnt3a、β-catenin蛋白及mRNA表达水平均降低(P值均<0.05),GPX4表达升高(P<0.05),病理损伤减轻;与实验组对比,联合组大鼠骨组织形态学发生一定程度改变,TNF-α、IL-6、COX-2、MMP-3、Wnt3a、β-catenin蛋白及mRNA表达水平均增高(P值均<0.05),GPX4表达降低(P<0.05),病理损伤较重。结论电针可激活OP大鼠Wnt/β-catenin通路,抑制骨质疏松铁死亡及炎症反应,改善OP大鼠骨损伤。

Objective To explore the therapeutic effect and mechanism of electroacupuncture rehabilitation intervention on the Wntβ-/catenin pathway in osteoporosis(OP)model rats.Methods Forty-eight female SD rats were randomly divided into blank group,model group,experimental group,and combination group(acupuncture+Wnt antagonist),with 12 rats in each group.Except for the blank group,the ovariectomy method was used in all other groups to establish the OP model rats.Rats in the experimental group received acupuncture treatment,while rats in the combined group received acupuncture treatment and Wnt antagonist DKK1 subcutaneous injection intervention.Rats in the other two groups were fed with conventional feed for 6 consecutive weeks.The morphological changes of bone tissue parameters between each group of rats were compared.ELISA was used to detect changes in serum TNF-α,IL-6,COX-2,and MMP-3.TUNEL method was used to analyze the apoptosis of femoral tissue cells.Immunohistochemistry was used to detect the expression of GPX4 protein.Western blotting and RT-PCR were used to detect the protein expressions of Wnt3a andβ-catenin and mRNA levels in cartilage tissue.Results Compared to that in the blank group,the bone tissue morphology in the model group rats showed significant changes,with increased levels of TNF-α,IL-6,COX-2,MMP-3,Wnt3a,andβ-catenin protein and mRNA expression(P<0.05)and decreased expression of GPX4(P<0.05),and severe pathological damage.Compared with the model group,the experimental group showed significant improvement in bone tissue morphology,with decreased levels of TNF-α,IL-6,COX-2,MMP-3,Wnt3a,β-catenin protein and mRNA expression(P<0.05)and increased expression of GPX4(P<0.05),and reduced pathological damage.Compared to that in the experimental group,the bone tissue morphology in the combined group rats underwent certain changes.The expression levels of TNF-α,IL-6,COX-2,MMP-3,Wnt3a,β-catenin protein and mRNA increased(P<0.05),GPX4 expression decreased(P<0.05),and pathological damage was more severe.Conclusion Acupuncture activates Wnt/β-catenin pathway and inhibits osteoporotic ferroptosis and inflammatory response,relieving bone damage in OP rats.