电针调节G6PD/NADPH/FAS/TKS5对佐剂性关节炎模型大鼠膝关节滑膜Th1、Th17相关促炎因子表达的影响

祁芳; 艾坤; 丁强盛; 赵君; 潘家慧; 吴庆泽; 瞿启睿; 张亮; 刘梨

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祁芳, 艾坤, 丁强盛, 赵君, 潘家慧, 吴庆泽, 瞿启睿, 张亮, 刘梨.电针调节G6PD/NADPH/FAS/TKS5对佐剂性关节炎模型大鼠膝关节滑膜Th1、Th17相关促炎因子表达的影响[J].湖南中医药大学学报英文版,2026,46(8):1629-1636.[Click to copy ]

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电针调节G6PD/NADPH/FAS/TKS5对佐剂性关节炎模型大鼠膝关节滑膜Th1、Th17相关促炎因子表达的影响

祁芳,艾坤,丁强盛,赵君,潘家慧,吴庆泽,瞿启睿,张亮,刘梨

(湖南中医药大学针灸推拿与康复学院, 湖南长沙 410208;湖南中医药大学第一附属医院, 湖南长沙 410007)

摘要:

目的以葡萄糖-6-磷酸脱氢酶（G6PD）/还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）/脂肪酸合成酶（FAS）代谢轴及其下游足体支架蛋白酪氨酸激酶底物（TKS5）为切入点，观察电针干预对佐剂性关节炎（AIA）大鼠关节滑膜内辅助性T（Th）1细胞、Th17细胞相关促炎因子表达的调控作用，初步阐释电针缓解类风湿关节炎（RA）滑膜炎症的分子作用机制。方法将36只SPF级SD大鼠按照体质量分层后，随机抽取9只设为空白组，余下27只先构建AIA模型，再随机分为模型组、药物组、电针组，每组9只。造模次日起，电针组每日行穴位电针干预，连续干预21 d；药物组予以甲氨蝶呤灌胃，每周2次，共干预6次。干预全程监测大鼠体质量与足跖容积；干预结束后，HE染色观察滑膜组织病理形态变化，免疫荧光染色检测滑膜组织中干扰素-γ（IFN-γ）、白细胞介素-17A（IL-17A）阳性表达以分别评估Th1、Th17浸润程度，qRT-PCR检测滑膜组织中TKS5 mRNA相对表达量，Western blot检测滑膜组织中G6PD、NADPH氧化酶 4（NOX4）、FAS蛋白相对表达量。结果干预结束后，与空白组相比，模型组大鼠足跖肿胀显著升高（P＜0.01），滑膜组织增生伴大量炎症细胞浸润，IFN-γ、IL-17A阳性表达率均显著上调（P＜0.01），TKS5 mRNA相对表达量及G6PD、NOX4、FAS蛋白相对表达量均显著升高（P＜0.01）；与模型组相比，电针组与药物组足跖肿胀程度均显著降低（P＜0.01），滑膜炎症浸润显著改善，IFN-γ、IL-17A阳性表达率均显著下调（P＜0.01），TKS5 mRNA相对表达量均显著降低（P＜0.01），G6PD、NOX4、FAS蛋白相对表达量均不同程度降低（P＜0.05，P＜0.01）。结论电针可有效改善AIA大鼠关节滑膜炎症，该效应可能通过调控G6PD/NADPH/FAS代谢轴及下游TKS5、减少促炎效应T细胞的滑膜浸润来实现。

DOI：10.3969/j.issn.1674-070X.2026.08.013

Received:May 21, 2026

基金项目:国家自然科学基金项目（82274627，82609199）；湖南省自然科学基金项目（2026JJ81029）；湖南中医药大学本科生科研创新基金项目（2025BKS218）。

Effects of electroacupuncture on expression of pro-inflammatory factors related to Th1 and Th17 in the kneesynovium of rats with adjuvant-induced arthritis by regulating the G6PD/NADPH/FAS/TKS5 axis

QI Fang, AI Kun, DING Qiangsheng, ZHAO Jun, PAN Jiahui, WU Qingze, QU Qirui, ZHANG Liang, LIU Li

(School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China)

Abstract:

Objective To investigate the regulatory effects of electroacupuncture (EA) on the expression of pro-inflammatory factors related to T helper (Th) 1 cells and Th17 cells, in the joint synovium of adjuvant-induced arthritis (AIA) rats, with the glucose-6-phosphate dehydrogenase (G6PD)/reduced nicotinamide adenine dinucleotide phosphate (NADPH)/fatty acid synthase (FAS) metabolic axis and its downstream podosome scaffold protein tyrosine kinase substrate 5 (TKS5) as the focus, and to preliminarily elucidate the molecular mechanism of EA in alleviating synovial inflammation in rheumatoid arthritis (RA). Methods Totally 36 SPF-grade SD rats were stratified by body weight and were randomly divided into a blank group (n=9) and the AIA model group (n=27). After successful establishment of the AIA models, the 27 rats were randomly divided into model, drug, and EA groups, with nine rats in each group. Beginning on the day after model establishment, the EA group received daily EA intervention at selected acupoints for 21 consecutive days, while the drug group was given methotrexate by gavage twice a week for a total of 6 interventions. Body weight and paw volume of rats were monitored throughout the intervention. After the intervention, HE staining was used to observe the pathomorphological changes in synovial tissue. Immunofluorescence staining was performed to determine the positive expression of interferon-γ (IFN-γ) and interleukin-17A (IL-17) A in synovial tissue to evaluate the infiltration of Th1 and Th17 cells, respectively. The relative mRNA expression levels of TKS5 and the relative protein expression levels of G6PD, NADPH oxidase 4(NOX4), and FAS in synovial tissue were determined by qRT-PCR and Western blot, respectively. Results After the intervention, compared with the blank group, paw swelling in rats of the model group increased significantly (P<0.01), with synovial hyperplasia accompanied by massive inflammatory cell infiltration, and significantly increased positive expression rates of IFN-γ and IL-17A (P<0.01), as well as significantly increased relative mRNA expression levels of TKS5 and the relative protein expression levels of G6PD, NOX4, and FAS (P<0.01). Compared with the model group, both the EA and drug groups showed a significantly reduced degree of paw swelling (P<0.01), alleviated synovial inflammatory infiltration, and significantly decreased positive expression rates of IFN-γ and IL-17A (P<0.01). The relative mRNA expression level of TKS5 decreased significantly in both groups (P<0.01), while the relative protein expression levels of G6PD, NOX4, and FAS declined to varying degrees (P<0.05, P<0.01). Conclusion EA can effectively ameliorate joint synovial inflammation in AIA rats, which may be associated with the regulation of the G6PD/NADPH/FAS metabolic axis and its downstream TKS5, thereby reducing the synovial infiltration of pro-inflammatory effector T cells.

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