热敏灸对骨质疏松小鼠破骨细胞PI3K/AKT/mTOR通路的影响  

Effect of thermal moxibustion on PI3K/AKT/mTOR pathway in osteoclasts of osteoporotic mice

作　　者：滕子忻 欧阳建江[2] 陈煌 王鹏[2] TENG Zixin;OUYANG Jianjiang;CHEN Huang;WANG Peng(Clinical Medical College,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004;Department of Sport Medicine and Arthroscopy,Jiangxi Provincial Hospital of Traditional Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]江西中医药大学附属医院,江西南昌330006

出　　处：《中国骨质疏松杂志》2024年第7期964-969,共6页Chinese Journal of Osteoporosis

基　　金：江西省中医药管理局科技计划重点项目(2021Z009)。

摘　　要：目的 探讨热敏灸对骨质疏松症(osteoporosis, OP)小鼠及破骨细胞PI3K/AKT/mTOR信号通路的干预效果。方法 雌性KM小鼠随机分为假手术组、模型组(双侧卵巢切除术)、雌激素组(0.205 5 mg/kg戊酸雌二醇灌胃)、艾灸组(悬灸肾俞穴,根据小鼠尾温变化分为热敏灸组和非热敏灸组)。干预8周后取材,使用双能X线骨密度仪测量小鼠股骨骨密度(bone mineral density, BMD)。通过RANKL体外诱导RAW264.7细胞,设为空白血清组、模型血清组、雌激素血清组、热敏灸血清组及非热敏灸血清组,分别加入10%对应组别小鼠血清。干预6 d后,采用TRAP染色检测破骨细胞分化情况,实时荧光定量PCR(qPCR)和蛋白免疫印迹法(Western blot)分别检测各组细胞中MMP-9、CTSK与p-PI3K、p-AKT、p-mTOR表达量。结果 与假手术组比较,模型组股骨BMD显著下降(P<0.05),雌激素组、热敏灸组、非热敏灸组股骨BMD较模型组显著升高。组间比较,热敏灸组小鼠的股骨BMD增加程度较雌激素组降低,较非热敏灸组升高(P<0.05)。血清干预6 d后,与空白血清组比较,模型血清组破骨细胞中MMP-9、CTSK、p-PI3K、p-AKT、p-mTOR表达水平上调,破骨细胞数显著增多(P<0.05)。与模型血清组比较,雌激素血清组、热敏灸血清组、非热敏灸血清组上述指标均显著逆转。组间比较,热敏灸血清组破骨细胞数较雌激素血清组增多,较非热敏灸血清组减少(P<0.05)。结论 热敏灸治疗OP小鼠可能是通过调控破骨细胞PI3K/AKT/mTOR信号通路抑制破骨细胞分化实现的,其干预效果优于传统艾灸。Objective To investigate the effect of thermal moxibustion on PI3K/AKT/mTOR signaling pathway in osteoporosis(OP)mice and osteoclasts.Methods Female KM mice were randomly divided into the sham operation group,model group(bilateral ovariectomy),estrogen group(0.2055 mg/kg estradiol valerate orally),moxibustion group(moxibustion at the Shenshu acupoint,dividing into thermal moxibustion group and non-thermal moxibustion group based on the tail temperature change of the mice).After 8 weeks of intervention,the femoral bone mineral density(BMD)of mice was measured with dual energy X-ray absorptiometry.RAW264.7 cells were induced by RANKL in vitro and divided into blank serum group,model serum group,estrogen serum group,thermal moxibustion serum group,and non-thermal moxibustion serum group,which were additioned with 10%serum of corresponding mice,respectively.After 6 days of intervention,osteoclast differentiation was detected with TRAP staining.MMP-9,CTSK,as well as p-PI3K,p-AKT,and p-mTOR expressions were detected with real-time fluorescent quantitative PCR(qPCR)and Western blotting in each cell group.Results Compared to those in the sham operation group,BMD in the model group decreased significantly(P<0.05),while BMD in the estrogen group,thermal moxibustion group,and non-thermal moxibustion group significantly increased.The increase degree of femoral BMD in the thermal moxibustion group was lower than that in the estrogen group,and higher than that in the non-thermal moxibustion group(P<0.05).After 6 days of serum intervention,compared to those in the blank serum group,the expression levels of CTSK,MMP-9,p-PI3K,p-AKT,and p-mTOR in osteoclasts from the model serum group were up-regulated,and the number of osteoclasts increased significantly(P<0.05).The above indexes in estrogen serum group,thermal moxibustion serum group and non-thermal moxibustion serum group were significantly reversed.The number of osteoclasts in the thermal moxibustion serum group was higher than that in the estrogen serum group,and lower than t

关 键 词：破骨细胞 热敏灸 骨质疏松症 PI3K/AKT/MTOR 信号通路 

分 类 号：R245.8[医药卫生—针灸推拿学] R589.5[医药卫生—中医临床基础] R-332[医药卫生—中医学]