Anti-inflammatory mechanism of acupoint acupuncture combined with tetramethylpyrazine in regulating the NF-κB/NLRP3 signaling pathway in a rat model of Kashin-Beck disease induced by T-2 toxin
10.3760/cma.j.cn231583-20250306-00087
- VernacularTitle:穴位针刺结合川芎嗪调控NF-κB/NLRP3信号通路对T-2毒素诱导的大骨节病大鼠模型抗炎作用机制研究
- Author:
Qi ZHANG
1
;
Xinlong MA
;
Haibo LI
;
Qian LIU
Author Information
1. 天津市天津医院（天津大学天津医院）中西医结合骨伤科，天津 300211
- Publication Type:Journal Article
- Keywords:
Kashin-Beck disease;
Acupoint acupuncture;
Tetramethylpyrazine;
Nuclear factor kappa B;
NOD like receptor thermal protein domain associated protein 3
- From:
Chinese Journal of Endemiology
2025;44(10):798-803
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the anti-inflammatory mechanism of acupoint acupuncture combined with tetramethylpyrazine in regulating the nuclear factor kappa B (NF-κB)/NOD like receptor thermal protein domain associated protein 3 (NLRP3) signaling pathway in a T-2 toxin induced rat model of Kashin-Beck disease.Methods:A total of 40 clean-grade Sprague-Dawley (SD) rats (half male and half female), weighing 180 - 220 g, were divided using a random number table method into model group, tetramethylpyrazine group, acupoint acupuncture combined with tetramethylpyrazine group, and control group, with 10 rats in each group. Rats in the model group, the tetramethylpyrazine group, and the acupoint acupuncture combined with tetramethylpyrazine group were intraperitoneally injected with 1.5 mg/kg of 0.1% T-2 toxin five times per week for 4 consecutive weeks. Rats in both the tetramethylpyrazine group and the acupoint acupuncture combined with tetramethylpyrazine group were intraperitoneally injected with 100 mg·kg -1·d -1 tetramethylpyrazine solution. Meanwhile, rats in the model group and the control group were administered an equivalent volume of physiological saline for consecutive 4 weeks. Acupoint acupuncture combined with tetramethylpyrazine group was used to needle the bilateral "Qu Chi" and "Wei Zhong" of rats with a diameter of 3 - 5 mm, leaving the needle for 20 minutes, once a day, for 4 weeks. After the last injection, the rats were euthanized to collect blood samples and knee joint cartilage. Hematoxylin and eosin staining was performed to observe the pathological changes in the knee joint cartilage of rats. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of osteocalcin (OC), alkaline phosphatase (ALP), β-carboxy-terminal cross-linked telopeptide of type Ⅰ collagen (β-CTX), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Western blot analysis was used to determine protein expression levels of NF-κB p65 and NLRP3. Results:In the control group, intact cartilage surface, middle, and deep structures were observed in the knee joint cartilage of rats. In the model group, pathological changes similar to adult Kashin-Beck disease were observed in the knee joint cartilage. The integrity of the knee joint cartilage of rats in the tetramethylpyrazine group and the acupoint acupuncture combined with tetramethylpyrazine group was significantly improved compared to the model group. Compared with the control group, the levels of OC and ALP in the serum of the model group rats were significantly reduced, while the level of β-CTX was significantly increased ( P < 0.05). Compared with the model group, the levels of OC and ALP in the serum of rats in the tetramethylpyrazine group and the acupoint acupuncture combined with tetramethylpyrazine group were significantly increased, while the levels of β-CTX were significantly decreased ( P < 0.05). The levels of OC and ALP in the serum of rats in the acupoint acupuncture combined with tetramethylpyrazine group were higher than those in the tetramethylpyrazine group, while the level of β-CTX was lower than that in the tetramethylpyrazine group ( P < 0.05). Compared with the control group, the levels of IL-1β, IL-6 and TNF-α in the serum of the model group rats were significantly increased ( P < 0.05). Compared with the model group, the levels of IL-1β, IL-6 and TNF-α in the serum of rats in the tetramethylpyrazine group and the acupoint acupuncture combined with tetramethylpyrazine group were significantly reduced ( P < 0.05). The levels of IL-1β, IL-6 and TNF-α in the serum of rats in the acupoint acupuncture combined with tetramethylpyrazine group were lower than those in the tetramethylpyrazine group ( P < 0.05). Compared with the control group, the expression levels of NF-κB p65 and NLRP3 proteins in the model group rats were significantly increased ( P < 0.05). Compared with the model group, the expression levels of NF-κB p65 and NLRP3 proteins were significantly reduced in the tetramethylpyrazine group and the acupoint acupuncture combined with tetramethylpyrazine group ( P < 0.05). The expression levels of NF-κB p65 and NLRP3 proteins in the acupoint acupuncture combined with tetramethylpyrazine group were significantly lower than those in the tetramethylpyrazine group ( P < 0.05). Conclusion:Acupoint acupuncture combined with tetramethylpyrazine has a good anti-inflammatory effect on T-2 toxin induced Kashin-Beck disease rats, and its mechanism may be related to the downregulation of NF-κB p65 and NLRP3 protein expression.
