Effect of heat-reinforcing needling on hypoxia-inducible factor 1α and glycolysis activity in rabbits with cold syndrome of rheumatoid arthritis.

Q3 Medicine

针刺研究 Pub Date : 2024-06-25 DOI:10.13702/j.1000-0607.20230487

Xiang-Jun Li, Fu-Xin Li, Wei-Yao Jing, Cui Liu, Li-Mei Liu, Xing-Hua Zhang, Feng-Fan Zhang, Ping Chen, Xiao-Zheng DU

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引用次数: 0

Abstract

Objectives: To observe the effect of heat-reinforcing needling (HRN) on synovial inflammation, hypoxia-inducible factor-1α (HIF-1α) and glycolytic activity in serum and synovial tissue in rabbits with cold syndrome of rheumatoid arthritis (RA), so as to explore its mechanisms underlying improvement of RA.

Methods: A total of 32 rabbits were randomly divided into normal, model, inhibitor and HRN groups, with 8 rabbits in each group. The RA with cold syndrome model was induced by injecting ovalbumin dry powder and Freund's complete adjuvant combined with cold freezing. Rabbits in the inhibitor group were intraperitoneally injected with 2-methoxyestradiol (2.5 mg/kg), rabbits in the HRN group were received HRN at bilateral "Zusanli" (ST36) for 30 min. The treatments were conducted once daily for 14 consecutive days. After the interventions, the knee circumference and pain threshold were measured. The contents of nicotinamide adenine dinucleotide phosphoric (NADPH), Hexokinase II (HK2) and 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3) in serum of rabbits were detected by ELISA. The pathological morphology of synovial tissue of the knee joints were observed by HE staining. The positive expressions of tumor necrosis factor (TNF-α), interleukin (IL)-1β, IL-6 and IL-17 in synovial tissue of knee joint were detected by immunohistochemistry. The content of lactic acid in synovial tissue of rabbit knee joint was detected by spectrophotometry. The expression levels of HIF-1α, pyruvate kinase 2 (PKM2) and lactate dehydrogenase (LDHA) in synovial tissue of knee joint were detected by Western blot.

Results: After intervention, compared with the normal group, the knee circumference was significantly enlarged (P<0.05), the pain threshold was significantly decreased (P<0.05)；the synovial tissue of knee joints showed significant cell proliferation and inflammatory infiltration, the pathological score was significantly increased (P<0.05)；positive expressions of TNF-α, IL-1β, IL-6 and IL-17, the content of lactic acid in synovial tissue, the contents of NADPH, HK2 and PFKFB3 in serum, and the protein expression levels of HIF-1α, PKM2 and LDHA in synovial tissue were increased (all P<0.05) in the model group. Compared with model group, the circumference of knee joint was significantly decreased (P<0.05), the pain threshold was significantly increased (P<0.05)；in synovial tissue, the pathological score was decreased (P<0.05)；the positive expressions of TNF-α, IL-1β, IL-6 and IL-17 in synovial tissue were decreased (P<0.05), the lactic acid content in synovial tissue was decreased (P<0.05)；the contents of NADPH, HK2 and PFKFB3 in serum and the protein expression levels of HIF-1α, PKM2 and LDHA in synovial tissue were decreased (P<0.05) in inhibitor group and HRN group. Compared with the inhibitor group, the synovial pathological score was significantly increased (P<0.05), positive expressions of TNF-α, IL-1β, IL-6 and IL-17, the content of lactic acid in synovial tissue, the contents of NADPH, HK2 and PFKFB3 in serum, and the protein expression levels of HIF-1α, PKM2 and LDHA in synovial tissue were increased (all P<0.05) in HRN group.

Conclusions: HRN can increase the pain threshold, reduce the knee circumference and inhibit the inflammatory response in rabbits with cold syndrome of RA. The possible mechanism is related to the down-regulation of HIF-1α and glycolysis activity.

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热强化针刺对类风湿性关节炎寒冷综合征家兔缺氧诱导因子 1α 和糖酵解活性的影响

目的观察热强化针刺（HRN）对类风湿性关节炎（RA）寒冷综合征家兔滑膜炎症、血清和滑膜组织中缺氧诱导因子-1α（HIF-1α）及糖酵解活性的影响，从而探讨其改善RA的机制：方法：将 32 只家兔随机分为正常组、模型组、抑制剂组和 HRN 组，每组 8 只。通过注射卵清蛋白干粉和弗氏完全佐剂并结合低温冷冻诱导RA伴寒冷综合征模型。抑制剂组的兔子腹腔注射 2-甲氧基雌二醇（2.5 毫克/千克），HRN 组的兔子接受双侧 "祖善利"（ST36）HRN 30 分钟。每天一次，连续 14 天。干预后，测量膝围和痛阈。用 ELISA 方法检测了兔子血清中烟酰胺腺嘌呤二核苷酸磷酸酶（NADPH）、己糖激酶 II（HK2）和 6-磷酸果糖-2-激酶/果糖-2，6-二磷酸酶 3（PFKFB3）的含量。通过 HE 染色法观察膝关节滑膜组织的病理形态。免疫组化法检测膝关节滑膜组织中肿瘤坏死因子（TNF-α）、白细胞介素（IL）-1β、IL-6 和 IL-17 的阳性表达。分光光度法检测兔膝关节滑膜组织中乳酸的含量。通过 Western blot 检测膝关节滑膜组织中 HIF-1α、丙酮酸激酶 2（PKM2）和乳酸脱氢酶（LDHA）的表达水平：结果：干预后，与正常组相比，膝关节周径明显增大：HRN能提高RA寒冷综合征家兔的痛阈，缩小膝围，抑制炎症反应。可能的机制与下调 HIF-1α 和糖酵解活性有关。

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来源期刊

针刺研究 Medicine-Medicine (all)

CiteScore

1.30

自引率

0.00%

发文量

期刊介绍： Acupuncture Research was founded in 1976. It is an acupuncture academic journal supervised by the State Administration of Traditional Chinese Medicine, co-sponsored by the Institute of Acupuncture of the China Academy of Chinese Medical Sciences and the Chinese Acupuncture Association. This journal is characterized by "basic experimental research as the main focus, taking into account clinical research and reporting". It is the only journal in my country that focuses on reporting the mechanism of action of acupuncture. The journal has been changed to a monthly journal since 2018, published on the 25th of each month, and printed in full color. The manuscript acceptance rate is about 10%, and provincial and above funded projects account for about 80% of the total published papers, reflecting the latest scientific research results in the acupuncture field and has a high academic level. Main columns: mechanism discussion, clinical research, acupuncture anesthesia, meridians and acupoints, theoretical discussion, ideas and methods, literature research, etc.