摘要
目的:探讨电针足少阳经穴通过调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路对绝经后骨质疏松大鼠炎性反应的影响。方法:40只SD大鼠随机选择10只作为假手术组;其余大鼠切除双侧卵巢3个月后构建绝经后骨质疏松模型,随机分为模型组、电针组、电针+抑制剂组,每组10只。电针组电针双侧“环跳”“阳陵泉”(疏密波,频率2 Hz/15 Hz,强度0.2~0.6 mA)并针刺“悬钟”,每次10 min,每周5次,共治疗8周,穴位左右交替。电针+抑制剂组在电针组治疗基础上腹腔注射PI3K抑制剂LY294002(10 mg/kg),每周2次,共治疗8周。Micro-CT检测大鼠股骨组织微结构及计量学指标,包括骨密度(BMD)、骨体积分数(BV/TV)、骨小梁数量(Tb.N)、骨小梁分离度(Tb.Sp)。HE染色观察股骨病理形态,抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞数量。免疫组织化学染色检测骨组织成骨标志物Runt相关转录因子2(Runx2)、骨钙素(OCN)、Ⅰ型胶原蛋白(CollagenⅠ)的表达。ELISA法及实时荧光定量PCR法分别检测血清及骨组织中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量及mRNA的表达。Western blot法检测股骨组织PI3K/AKT通路蛋白表达。结果:模型组大鼠骨小梁明显变细,排列稀疏、紊乱,骨髓腔扩大,破骨细胞数量明显增多;电针组的骨小梁结构形态改善,破骨细胞数量减少。与假手术组相比,模型组大鼠BMD 、BV/TV 、Tb.N,骨组织Runx2、OCN 、CollagenⅠ阳性表达,p-PI3K/PI3K、p-AKT/AKT蛋白表达比值显著降低(P<0.01),Tb.Sp、血清及骨组织中的IL-6、IL-1β、TNF-α含量及mRNA表达显著升高(P<0.01)。与模型组相比,电针组大鼠BMD、BV/TV、Tb.N,骨组织Runx2、OCN、CollagenⅠ阳性表达,p-PI3K/PI3K、p-AKT/AKT蛋白表达比值显著升高(P<0.01,P<0.05),Tb.Sp、血清及骨组织中的IL-6、IL-1β、TNF-α含量及mRNA表达显著降低(P<0.01)。与电针组相比,电针+抑制剂组大鼠上述指标除Tb.Sp外均逆转(P<0.01,P<0.05)。结论:电针足少阳经穴可减轻绝经后骨质疏松大鼠的炎性反应和骨质流失,其作用机制可能与PI3K/AKT通路的激活有关。
Objective To investigate the effect of electroacupuncture(EA)at Foot Shaoyang meridian acupoints on the inflammatory response in postmenopausal osteoporosis rats by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods Forty Sprague-Dawley(SD)rats were randomly divided into a sham operation group(n=10),and the remaining rats underwent bilateral ovariectomy to establish a postmenopausal osteoporosis model after 3 months.The model rats were further randomized into a model group,an EA group,and an EA+inhibitor group(n=10).Rats of The EA group received EA at bilateral“Huantiao”(GB30),“Yanglingquan”(GB34)(dense-sparse wave,2 Hz/15 Hz,0.2—0.6 mA),and acupuncture at“Xuanzhong”(GB39),10 min each time,5 times a week for 8 weeks,with alternate left-right acupoints.Rats of the EA+inhibitor group received intraperitoneal injection of the PI3K inhibitor LY294002(10 mg/kg)twice a week for 8 weeks based on the EA treatment.Micro-CT was used to detect the microstructural and morphometric indices of the femoral bone tissue,including bone mineral density(BMD),bone volume fraction(BV/TV),trabecular number(Tb.N),and trabecular separation(Tb.Sp).HE staining was performed to observe the pathological morphology of the femoral tissue,and tartrate-resistant acid phosphatase(TRAP)staining was used to observe the number of osteoclasts.Immunohistochemical staining was employed to detect the expressions of osteogenic markers Runt-related transcription factor 2(Runx2),osteocalcin(OCN),and type I collagen(Collagen I).The contents of serum interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α),as well as their mRNA expressions in bone tissue,were detected by ELISA and real-time fluorescent quantitative PCR,respectively.Western blot was used to detect the protein expressions of the PI3K/AKT pathway in femoral tissue.Results In the model group,bone trabeculae were significantly thinned,sparsely and disorderly arranged,the bone marrow cavity was expanded,and the number of osteoclasts was significantly increased;the EA group showed improved bone trabecular structure and reduced osteoclast number.Compared with the sham operation group,the model group had significantly lower BMD,BV/TV,Tb.N,positive expressions of Runx2,OCN,and Collagen I in bone tissue,and ratios of p-PI3K/PI3K and p-AKT/AKT protein expression(P<0.01),while Tb.Sp,serum IL-6,IL-1β,TNF-αcontents and their mRNA expressions in bone tissue were significantly higher(P<0.01).Compared with the model group,the EA group had significantly higher BMD,BV/TV,Tb.N,positive expressions of Runx2,OCN,and Collagen I in bone tissue,and ratios of p-PI3K/PI3K and p-AKT/AKT protein expression(P<0.01,P<0.05),while Tb.Sp,serum IL-6,IL-1β,TNF-αcontents and their mRNA expressions in bone tissue were significantly lower(P<0.01).Compared with the EA group,all the above indices except Tb.Sp were reversed in the EA+inhibitor group(P<0.01,P<0.05).Conclusion EA at Foot Shaoyang meridian acupoints can alleviate inflammatory responses and bone loss in postmenopausal osteoporosis rats,and its mechanism may be related to the activation of the PI3K/AKT pathway.
作者
吕兴民
唐偲
荣熠
邹玲玲
徐茂婷
张璐
陈贵全
LÜXing-min;TANG Cai;RONG Yi;ZOU Ling-ling;XU Mao-ting;ZHANG Lu;CHEN Gui-quan(Department of Acupuncture,Tuina and Rehabilitation,Traditional Chinese Medicine Hospital Affiliated to Southwest Medical University,Luzhou 646000,Sichuan Province,China;College of Integrated Traditional Chinese and Western Medicine,Southwest Medical University,Luzhou 646000,Sichuan Province)
出处
《针刺研究》
北大核心
2026年第1期96-105,共10页
Acupuncture Research
基金
西南医科大学校级科研项目(No.2021ZKZD020)
第五批全国中医临床优秀人才研修项目资助(No.国中医药人教函[2022]1号)。
关键词
电针
足少阳经穴
磷脂酰肌醇3-激酶/蛋白激酶B通路
骨质疏松
炎性反应
Electroacupuncture
Foot Shaoyang meridian acupoints
Phosphatidylinositol 3-kinase/protein kinase B pathway
Osteoporosis
Inflammatory response
