基于线粒体能量代谢探讨刺络拔罐法对内毒素致热家兔退热作用的影响

Effect of Pricking-Cupping Therapy on Endotoxin-Induced Fever in Rabbits Based on Mitochondrial Energy Metabolism

    摘要
  • 摘要:
    目的 通过观察刺络拔罐法对内毒素致热家兔的退热作用，从骨骼肌线粒体能量代谢的角度探讨刺络拔罐法的退热机制。
    方法 将72只家兔随机分为空白组、模型组、西药组、刺络组、拔罐组和刺络拔罐组，每组12只；复制内毒素致热家兔模型，并进行相应治疗干预。连续6 h记录家兔体温；利用透射电镜观察线粒体结构；采用Oxygraph-2k系统检测线粒体功能；检测各组家兔大腿肌肉组织中ATP含量；采用Wes全自动蛋白质印迹定量分析系统检测家兔大腿肌肉组织解偶联蛋白2(UCP2)、线粒体融合蛋白2(Mfn2)蛋白表达。
    结果 模型复制成功，4个治疗组家兔体温均低于模型组；空白组家兔骨骼肌线粒体形态完整，其中经刺络拔罐法治疗后线粒体形态最接近正常；刺络拔罐组线粒体CII OXPHOS阶段耗氧量较模型组明显升高；与模型组相比，刺络拔罐组家兔肌肉组织ATP含量显著升高；与模型组比较，刺络拔罐组肌肉组织Mfn2蛋白表达显著升高，UCP2蛋白表达明显降低。
    结论 刺络拔罐法对内毒素致热家兔具有良好的退热效果，其机制可能与抑制骨骼肌中UCP2蛋白表达，促进Mfn2蛋白表达，推动骨骼肌内线粒体氧化磷酸化进程，增加线粒体内ATP合成相关。

     

    Abstract:
    OBJECTIVE To observe the fever-reducing effect of pricking-cupping method on endotoxin-induced fever in rabbits, and to further explore the fever-reducing mechanism of pricking-cupping method from the perspective of skeletal muscle mitochondrial energy metabolism.
    METHODS A total of 72 rabbits were randomly divided into blank group, model group, Western medicine group, pricking group, cupping group and pricking-cupping group, with 12 rabbits in each group.The rabbit model of endotoxin-induced fever was reproduced and the corresponding therapeutic intervention was carried out.The body temperature of rabbits was recorded continuously for 6 h.The mitochondrial structure was observed by transmission electron microscopy.Oxygraph-2k system was used to detect mitochondrial function.ATP content in thigh muscle tissue of rabbits in each group was detected.The expression of uncoupling protein 2 (UCP2) and mitochondrial fusion protein 2 (Mfn2) in thigh muscle of rabbits was detected by Wes automated western blot quantitative analysis system.
    RESULTS The model was successfully replicated, and the body temperature of rabbits in the four treatment groups was lower than that in the model group. The skeletal muscle mitochondrial morphology of rabbits in the blank group was intact, and the mitochondrial morphology was closest to normal after the treatment of pricking-cupping. Moreover, the oxygen consumption of mitochondrial CII OXPHOS stage in the pricking-cupping group was significantly higher than that in the model group. Compared with the model group, the ATP content in the muscle tissue of rabbits in the pricking-cupping group was significantly increased, Mfn2 protein expression was significantly increased, and UCP2 protein expression was significantly decreased.
    CONCLUSION The pricking-cupping method has a good fever-reducing effect on endotoxin-induced fever in rabbits, and its mechanism may be related to inhibiting UCP2 protein expression in skeletal muscle, promoting Mfn2 protein expression, promoting the oxidative phosphorylation process in skeletal muscle mitochondria, and increasing ATP synthesis in mitochondria.

     

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