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Regulatory effect of mild moxibustion on P2X3 receptors in spinal cord, anterior cingulate cortex and thalamic ventral posterolateral nucleus of rats with IBS visceral hyperalgesia

温和灸对IBS内脏痛敏大鼠脊髓、 前扣带回和丘脑腹后外侧核内P2X3受体的调节作用

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Abstract

Objective

To observe the therapeutic effect of mild moxibustion on irritable bowel syndrome (IBS) visceral hyperalgesia model rats and its regulatory effect on P2X3 receptors in the spinal cord, anterior cingutate cortex (ACC) and thalamic ventral posterolateral nucleus (VPL).

Methods

Thirty 8-day-old newborn rats were randomly divided into a normal group (n=6) and a modeling group (n=24) according to the completely random number table method. Rats in the normal group were bred routinely, and those in the modeling group were subjected to preparing IBS chronic visceral hyperalgesia model using colorectal distention (CRD) in stimulation method. Rats successfully modelled were re-divided into a model group, a mild moxibustion group, a P2X3 receptor antagonist group, and a normal saline group according to the completely random number table method with 6 rats in each group. Rats in each group received corresponding interventions from the 37-day old, once a day for 7 consecutive days. Immunohistochemistry and Western blot assays were used to detect P2X3 protein expressions in the spinal cord, ACC and VPL of rats.

Results

Under different intensities of CRD stimulation, the abdominal withdrawal reflex (AWR) scores of the model group were significantly increased versus the normal group (all P<0.05); the AWR scores of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group (all P<0.01). The P2X3 protein expressions in rat spinal cord, ACC and VPL tissues of the model group were significantly increased versus the normal group (all P<0.01); the P2X3 protein expressions in rat spinal cord, ACC and VPL tissues of the mild moxibustion group and the P2X3 receptor antagonist group were significantly reduced versus the model group (all P<0.01).

Conclusion

Mild moxibustion can inhibit the P2X3 receptor expressions in the spinal cord, ACC, and VPL tissues of IBS visceral hyperalgesia model rats, which may be the mechanism of mild moxibustion in relieving the central sensitization of rats with IBS visceral hyperalgesia.

摘要

目的

观察温和灸对肠易激综合征(IBS)内脏痛敏模型大鼠的干预作用及其对脊髓、 前扣带回(ACC)和丘脑腹后外侧核(VPL)中P2X3受体的调节作用.

方法

按照完全随机数字表法将30只8日龄新生大鼠随机分为正常组(n=6)与造模组(n=24). 正常组常规饲养, 造模组采用直结肠球囊扩张(CRD)刺激法制备IBS慢性内脏痛敏大鼠模型. 将模型验证成功的造模大鼠按照完全随机数字表法重新分为模型组、 温和灸组、 P2X3受体拮抗剂组和生理盐水组, 每组6只. 各组大鼠从37日龄开始分别接受相应的干预, 每日1次, 连续7 d. 采用免疫组织化学法及免疫印迹法检测大鼠脊髓、 ACC和VPL内P2X3蛋白表达.

结果

在不同强度的CRD刺激下, 与正常组相比, 模型组AWR评分均明显升高(均P<0.05); 与模型组相比, 温和灸组及P2X3受体拮抗剂组AWR评分均显著降低(均P<0.01). 与正常组相比, 模型组大鼠脊髓、 ACC及VPL组织中P2X3蛋白的表达均显著升高(均P<0.01); 与模型组相比, 温和灸组及P2X3受体拮抗剂组大鼠脊髓、 ACC及VPL组织中P2X3蛋白的表达均显著降低(均P<0.01).

结论

温和灸能够抑制IBS内脏痛敏模型大鼠脊髓、 ACC及VPL组织内P2X3受体的表达, 这可能是温和灸缓解IBS内脏痛敏大鼠中枢敏化的效应机制.

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Acknowledgments

This work was supported by Shanghai Municipal Commission of Health and Family Planning (上海市卫生 和计划生育委员会科研课题, No. 20174Y0015); National Natural Science Foundation of China (国家自然科学基金 项目, No. 81904301); National Basic Research Program of China (973 Program, 国家重点基础研究发展计划项目 No. 2015CB554501); Three-year Action Plan for the Development of Shanghai Traditional Chinese Medicine [上 海中医药事业发展三年行动计划项目, No. ZY(2018–2020)-CCCX-2004-01]; National Chinese Medicine Leading Talent Support Program-Qi Huang Scholar (国家 中医药领军人才支持计划-岐黄学者).

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Correspondence to Hui-rong Liu  (刘慧荣) or Yun-hua Cui  (崔云华).

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Author Wu Huan-gan is a member of the Editorial Board of Journal of Acupuncture and Tuina Science. The paper was handled by other editors and has undergone rigorous peer review process. Author Wu Huan-gan was not involved in the journal’s review or decisions related to this manuscript.

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Statement of Human and Animal Rights

The treatment of animals conformed to the ethical criteria in this experiment.

Co-first Authors: Zhang Zhi-ying, master degree candidate; Zhang Fang, M.M., research intern

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Zhang, Zy., Zhang, F., Weng, Zj. et al. Regulatory effect of mild moxibustion on P2X3 receptors in spinal cord, anterior cingulate cortex and thalamic ventral posterolateral nucleus of rats with IBS visceral hyperalgesia. J. Acupunct. Tuina. Sci. 19, 239–248 (2021). https://doi.org/10.1007/s11726-021-1254-8

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