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林巧婷, 钟叶蓓, 杨尚林, 裴鑫, 杨小慧, 李铁浪, 李武.基于α7nAChR/NF-κB通路研究点按脾俞、胃俞穴对缓解慢性疲劳综合征大鼠外周炎症的效应机制[J].湖南中医药大学学报,2024,44(6):1027-1033[点击复制] |
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| 基于α7nAChR/NF-κB通路研究点按脾俞、胃俞穴对缓解慢性疲劳综合征大鼠外周炎症的效应机制 |
| 林巧婷,钟叶蓓,杨尚林,裴鑫,杨小慧,李铁浪,李武 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察点按脾俞、胃俞穴对慢性疲劳综合征（chronic fatigue syndrome, CFS）模型大鼠疲劳状态、运动行为、外周炎症及α7烟碱型乙酰胆碱受体（α7 nicotinic acetylcholine receptor, α7nAChR）/核因子κB（nuclear factor-κB, NF-κB）信号表达的影响，探讨点按脾俞、胃俞穴对缓解CFS大鼠外周炎症的效应机制。方法 先将32只SD大鼠随机分为空白组（8只）和造模组（24只）。造模组采用强迫负重游泳联合慢性应激刺激的方法建立CFS大鼠模型。造模成功后，将造模组大鼠随机分为模型组、点按组、α7nAChR激动剂组，每组8只。空白组、模型组予腹腔注射等体积生理盐水；点按组以自制按法刺激仪点按双侧脾俞、胃俞穴，20 min/次，并予腹腔注射等体积生理盐水；α7nAChR激动剂组予腹腔注射α7nAChR激动剂PNU-282987（每次2.4 mg/kg）。每组每日干预1次，连续14 d。记录各组大鼠一般情况半定量评分、力竭游泳时间和旷场实验运动距离；计算大鼠脾脏指数和胸腺指数；采用ELISA法检测大鼠血清中肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素-1β（interleukin-1β, IL-1β）、白细胞介素-6（interleukin-6, IL-6）的含量；Western blot检测大鼠脾脏组织α7nAChR、NF-κB p65、NF-κB p-p65、TNF-α的蛋白表达水平；采用qPCR法检测α7nAChR、NF-κB p65、TNF-α的mRNA表达水平。结果 与空白组比较，模型组一般情况半定量评分，血清TNF-α、IL-1β、IL-6含量，脾脏组织中NF-κB p-p65、NF-κB p65、TNF-α蛋白表达水平及NF-κB p65、TNF-α mRNA表达量均升高（P<0.01）；力竭游泳时间、旷场实验运动距离、脾脏指数、胸腺指数、脾脏组织中α7nAChR蛋白表达水平及mRNA表达量均降低（P<0.05，P<0.01）。与模型组比较，点按组与α7nAChR激动剂组一般情况半定量评分，血清TNF-α、IL-1β、IL-6含量，脾脏组织中NF-κB p-p65、NF-κB p65、TNF-α蛋白表达水平及NF-κB p65、TNF-α mRNA表达量均降低（P<0.05，P<0.01）；力竭游泳时间、旷场实验运动距离、脾脏指数、胸腺指数、脾脏组织中α7nAChR蛋白表达水平及mRNA表达量均升高（P<0.05，P<0.01）。与点按组比较，α7nAChR激动剂组血清TNF-α、IL-1β、IL-6含量，脾脏组织中NF-κB p-p65、NF-κB p65、TNF-α蛋白表达水平及NF-κB p65、TNF-α mRNA表达量均降低（P<0.01），脾脏组织中α7nAChR蛋白表达水平及mRNA表达量均升高（P<0.05，P<0.01）。结论 点按脾俞、胃俞穴可有效改善CFS大鼠的疲劳状态和运动行为，提高免疫水平并缓解外周炎症，这可能与激活α7nAChR的表达，从而抑制NF-κB下游炎性通路的活性相关。 |
| 关键词: 慢性疲劳综合征 点按法 脾俞 胃俞 α7烟碱型乙酰胆碱受体/核转录因子κB通路 外周炎症 |
| DOI：10.3969/j.issn.1674-070X.2024.06.013 |
| 投稿时间：2023-12-21 |
| 基金项目:湖南省自然科学基金项目（2023JJ30454）；湖南省教育厅科学研究重点项目（22A0273）；湖南中医药大学研究生创新课题项目（2022CX107）。 |
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| Mechanism of effects of pressing Pishu (BL20) and Weishu (BL21) on alleviating peripheral inflammation in rats with chronic fatigue syndrome based on the α7nAChR/NF-κB pathway |
| LIN Qiaoting, ZHONG Yebei, YANG Shanglin, PEI Xin, YANG Xiaohui, LI Tielang, LI Wu |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effects of pressing Pishu (BL20) and Weishu (BL21) on the fatigue state, locomotor behaviors, peripheral inflammation, and α7 nicotinic acetylcholine receptor (α7nAChR)/nuclear factor-κB (NF-κB) signaling expression of chronic fatigue syndrome (CFS) model rats, and to explore the mechanism of effect of this therapy on the alleviation of peripheral inflammation in CFS rats. Methods A total of 32 SD rats were randomized into blank group (n=8) and modeling group (n=24). In the modeling group, a CFS model was established by forced weight-bearing swimming combined with chronic stress stimulation. After successful modeling, the rats in the modeling group were randomly subdivided into model, acupoint-pressing, and α7nAChR agonist groups, with eight rats in each group. The blank and model groups were injected intraperitoneally with an equal volume of saline; the acupoint-pressing group was subjected to the bilateral press on Pishu (BL20) and Weishu (BL21) using a homemade pressing stimulator for 20 min each time as well as intraperitoneal injection of an equal volume of saline; the α7nAChR agonist group was injected intraperitoneally with the α7nAChR agonist PNU-282987 (2.4 mg/kg per time). Each group was intervened once daily for continuous 14 days. The semi-quantitative score of the general condition, swimming time to exhaustion, and exercise distance in the open-field test were recorded in each group; the spleen index and thymus index were calculated; the content of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in rat serum were checked using ELISA; the protein expression levels of α7nAChR, NF-κB p65, NF-κB p-p65, and TNF-α in the rat spleen tissue were examined by Western blot; the mRNA expression levels of α7nAChR, NF-κB p65, and TNF-α were determined by qPCR. Results Compared with the blank group, the semi-quantitative score of the general condition, serum content of TNF-α, IL-1β, and IL-6, as well as the protein expression levels of NF-κB p-p65, NF-κB p65, and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue increased in the model group (P<0.01); the swimming time to exhaustion and exercise distance in the open-field test were shortened, and the spleen index, thymus index, and the α7nAChR protein and mRNA expression levels in the spleen tissue decreased (P<0.05, P<0.01). Compared with the model group, the semi-quantitative scores of the general condition, serum content of TNF-α, IL-1β, and IL-6, as well as the protein expression levels of NF-κB p-p65, NF-κB p65, and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue were reduced in both acupoint-pressing and α7nAChR agonist groups (P<0.05, P<0.01); the swimming time to exhaustion and exercise distance in the open-field test were longer, and the spleen index, thymus index, and the α7nAChR protein and mRNA expression levels in the spleen tissue were higher (P<0.05, P<0.01). Compared with the acupoint-pressing group, the serum content of TNF-α, IL-1β, and IL-6, as well as the protein expression levels of NF-κB p-p65, NF-κB p65, and TNF-α and the mRNA expression levels of NF-κB p65 and TNF-α in the spleen tissue were lower in the α7nAChR agonist group (P<0.01), while the α7nAChR protein and mRNA expression levels in the spleen tissue were elevated (P<0.05, P<0.01). Conclusion Pressing Pishu (BL20) and Weishu (BL21) can effectively relieve fatigue, improve locomotor behaviors, enhance immune level, and alleviate peripheral inflammation in CFS rats, which may be related to the activation of the expression of α7nAChR, thereby inhibiting the activity of the downstream inflammatory pathway of NF-κB. |
| Key words: chronic fatigue syndrome pressing Pishu (BL20) Weishu (BL21) α7 nicotinic acetylcholine receptor/nuclear factor-κB pathway peripheral inflammation |
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