Skip to main content

Advertisement

Log in

Electro-acupuncture Pretreatment at Zusanli (ST36) Acupoint Attenuates Lipopolysaccharide-Induced Inflammation in Rats by Inhibiting Ca2+ Influx Associated with Cannabinoid CB2 Receptors

  • ORIGINAL ARTICLE
  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

In this study, we aimed to investigate the effect of electro-acupuncture (EA) pretreatment at zusanli (ST36) acupoint on lipopolysaccharide (LPS)-induced endotoxemic rat model and explore the underlying molecular mechanisms. Rats were treated with EA at ST36 for 7 days before being subjected to LPS. Two hours post-LPS, samples such as serum, local acupoint tissues, and spleens were collected and processed for investigations including cytokine production, cytosolic calcium (Ca2+) concentration, Ca2+ influx, cannabinoid CB2 receptor (CB2R) expression, and TLR4/NF-κB signaling. Our results showed EA pretreatment significantly attenuated LPS-induced inflammatory cytokine production, such as TNF-α, IL-1β, and IL-6. EA also enhanced CB2R expression, inhibited Ca2+ influx, and inactivated TLR4/NF-κB signaling, subsequently resulting in a substantial reduction of Ca2+ concentration. Importantly, CB2R antagonist AM630 effectively abrogated the suppressive effect of EA at ST36 on the endotoxemic rats, suggesting CB2R was involved in the anti-inflammatory effect of EA. EA pretreatment could enhance CB2R expression, inhibit Ca2+ influx, and inactivate TLR4/NF-κB signaling, which contributes to the alleviation of LPS-induced inflammation in rats.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Tracey, K.J. 2002. The inflammatory reflex. Nature 420: 853–859.

    Article  PubMed  CAS  Google Scholar 

  2. Schett, G., D. Elewaut, I.B. McInnes, J.M. Dayer, and M.F. Neurath. 2013. How cytokine networks fuel inflammation: toward a cytokine-based disease taxonomy. Nature Medicine 19: 822–824.

    Article  PubMed  CAS  Google Scholar 

  3. Headland, S.E., and L.V. Norling. 2015. The resolution of inflammation: principles and challenges. Seminars in Immunology 27: 149–160.

    Article  PubMed  CAS  Google Scholar 

  4. Raetz, C.R., and C. Whitfield. 2002. Lipopolysaccharide endotoxins. Annual Review of Biochemistry 71: 635–700.

    Article  PubMed  CAS  Google Scholar 

  5. Płóciennikowska, A., A. Hromada-Judycka, K. Borzęcka, and K. Kwiatkowska. 2015. Co-operation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling. Cellular and Molecular Life Sciences 72: 557–581.

    Article  PubMed  CAS  Google Scholar 

  6. Opal, S.M. 2010. Endotoxins and other sepsis triggers. Contributions to Nephrology 167: 14–24.

    Article  PubMed  CAS  Google Scholar 

  7. Xue, X., Y. You, J. Tao, X. Ye, J. Huang, S. Yang, Z. Lin, Z. Hong, J. Peng, and L. Chen. 2014. Electro-acupuncture at points of zusanli and quchi exerts anti-apoptotic effect through the modulation of PI3K/Akt signaling pathway. Neuroscience Letters 558: 14–19.

    Article  PubMed  CAS  Google Scholar 

  8. Lu, Y., H. Zhao, Y. Wang, B. Han, T. Wang, H. Zhao, K. Cui, and S. Wang. 2015. Electro-acupuncture up-regulates astrocytic MCT1 expression to improve neurological deficit in middle cerebral artery occlusion rats. Life Sciences 134: 68–72.

    Article  PubMed  CAS  Google Scholar 

  9. Youn, J.I., K.K. Sung, B.K. Song, M. Kim, and S. Lee. 2013. Effects of electro-acupuncture therapy on post-stroke depression in patients with different degrees of motor function impairments: a pilot study. Journal of Physical Therapy Science 25: 725–728.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Fukazawa, Y., T. Maeda, and S. Kishioka. 2009. The pharmacological mechanisms of electroacupuncture. Current Opinion in Investigational Drugs 10: 62–69.

    PubMed  Google Scholar 

  11. Wang, Z., T. Chen, M. Long, L. Chen, L. Wang, N. Yin, and Z. Chen. 2017. Electro-acupuncture at acupoint ST36 ameliorates inflammation and regulates Th1/Th2 balance in delayed-type hypersensitivity. Inflammation 40: 422–434.

    Article  PubMed  CAS  Google Scholar 

  12. Wang, Z., T. Yi, M. Long, Y. Gao, C. Cao, C. Huang, Q. Wang, N. Yin, and Z. Chen. 2017. Electro-acupuncture at zusanli acupoint (ST36) suppresses inflammation in allergic contact dermatitis via triggering local IL-10 production and inhibiting p38 MAPK activation. Inflammation 40: 1351–1364.

    Article  PubMed  CAS  Google Scholar 

  13. Liu, H.W., M.C. Liu, C.M. Tsao, M.H. Liao, and C.C. Wu. 2011. Electro-acupuncture at ‘Neiguan’ (PC6) attenuates liver injury in endotoxaemic rats. Acupuncture in Medicine 29: 284–288.

    Article  PubMed  Google Scholar 

  14. Zhang, Y., J.B. Yu, X.Q. Luo, L.R. Gong, M. Wang, X.S. Cao, S.A. Dong, Y.M. Yan, Y. Kwon, and J. He. 2014. Effect of ERK1/2 signaling pathway in electro-acupuncture mediated up-regulation of heme oxygenase-1 in lungs of rabbits with endotoxic shock. Medical Science Monitor 20: 1452–1460.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Liu, Z., X. Chen, Y. Gao, S. Sun, L. Yang, Q. Yang, F. Bai, L. Xiong, and Q. Wang. 2015. Involvement of GluR2 up-regulation in neuroprotection by electroacupuncture pretreatment via cannabinoid CB1 receptor in mice. Scientific Reports 5: 9490.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. Schmittgen, T.D., and K.J. Livak. 2008. Analyzing real-time PCR data by the comparative C(T) method. Nature Protocols 3: 1101–1108.

    Article  PubMed  CAS  Google Scholar 

  17. Yin, N., X. Hong, Y. Han, Y. Duan, Y. Zhang, and Z. Chen. 2015. Cortex Mori Radicis extract induces neurite outgrowth in PC12 cells activating ERK signaling pathway via inhibiting Ca(2+) influx. International Journal of Clinical and Experimental Medicine 8: 5022–5032.

    PubMed  PubMed Central  Google Scholar 

  18. Schappe, M.S., K. Szteyn, M.E. Stremska, S.K. Mendu, T.K. Downs, P.V. Seegren, M.A. Mahoney, S. Dixit, J.K. Krupa, E.J. Stipes, J.S. Rogers, S.E. Adamson, N. Leitinger, and B.N. Desai. 2018. Chanzyme TRPM7 mediates the Ca2+ influx essential for lipopolysaccharide-induced Toll-like receptor 4 endocytosis and macrophage activation. Immunity 48: 59–74.e5.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Zhang, Y., Y. Lu, L. Ma, X. Cao, J. Xiao, J. Chen, S. Jiao, Y. Gao, C. Liu, Z. Duan, D. Li, Y. He, B. Wei, and H. Wang. 2014. Activation of vascular endothelial growth factor receptor-3 in macrophages restrains TLR4-NF-κB signaling and protects against endotoxin shock. Immunity 40: 501–514.

    Article  PubMed  CAS  Google Scholar 

  20. Olmos, G., and J. Lladó. 2014. Tumor necrosis factor alpha: a link between neuroinflammation and excitotoxicity. Mediators of Inflammation 2014: 861231.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  21. Song, Q., S. Hu, H. Wang, Y. Lv, X. Shi, Z. Sheng, and W. Sheng. 2014. Electroacupuncturing at zusanli point (ST36) attenuates pro-inflammatory cytokine release and organ dysfunction by activating cholinergic anti-inflammatory pathway in rat with endotoxin challenge. African Journal of Traditional, Complementary, and Alternative Medicines 11: 469–474.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  22. Yu, J.B., S.A. Dong, X.Q. Luo, L.R. Gong, Y. Zhang, M. Wang, X.S. Cao, and D.Q. Liu. 2013. Role of HO-1 in protective effect of electro-acupuncture against endotoxin shock-induced acute lung injury in rabbits. Experimental Biology and Medicine (Maywood, N.J.) 238: 705–712.

    Article  CAS  Google Scholar 

  23. Park, J.Y., and U. Namgung. 2018. Electroacupuncture therapy in inflammation regulation: current perspectives. Journal of Inflammation Research 11: 227–237.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  24. Jung, T.G., J.H. Lee, I.S. Lee, and B.T. Choi. 2010. Involvement of intracellular calcium on the phosphorylation of spinal N-methyl-d-aspartate receptor following electroacupuncture stimulation in rats. Acta Histochemica 112: 127–132.

    Article  PubMed  CAS  Google Scholar 

  25. Gao, J., Y. Zhao, Y. Wang, J. Xin, J. Cui, S. Ma, F. Lu, L. Qin, and X. Yu. 2015. Anti-arrhythmic effect of acupuncture pretreatment in the rats subjected to simulative global ischemia and reperfusion—involvement of intracellular Ca2+ and connexin 43. BMC Complementary and Alternative Medicine 15: 5.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  26. Chen, L., A. Xu, N. Yin, M. Zhao, Z. Wang, T. Chen, Y. Gao, and Z. Chen. 2017. Enhancement of immune cytokines and splenic CD4+ T cells by electroacupuncture at ST36 acupoint of SD rats. PLoS One 12: e0175568.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  27. Mittal, M., C. Tiruppathi, S. Nepal, Y.Y. Zhao, D. Grzych, D. Soni, D.J. Prockop, and A.B. Malik. 2016. TNFα-stimulated gene-6 (TSG6) activates macrophage phenotype transition to prevent inflammatory lung injury. Proceedings of the National Academy of Sciences of the United States of America 113: E8151–E8158.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  28. Zhou, X., W. Yang, and J. Li. 2006. Ca2+- and protein kinase C-dependent signaling pathway for nuclear factor-kappaB activation, inducible nitric-oxide synthase expression, and tumor necrosis factor-alpha production in lipopolysaccharide-stimulated rat peritoneal macrophages. The Journal of Biological Chemistry 281: 31337–31347.

    Article  PubMed  CAS  Google Scholar 

  29. Toczek, M., and B. Malinowska. 2018. Enhanced endocannabinoid tone as a potential target of pharmacotherapy. Life Sciences 204: 20–45.

    Article  PubMed  CAS  Google Scholar 

  30. Chen, L., J. Zhang, F. Li, Y. Qiu, L. Wang, Y.H. Li, J. Shi, H.L. Pan, and M. Li. 2009. Endogenous anandamide and cannabinoid receptor-2 contribute to electroacupuncture analgesia in rats. The Journal of Pain 10: 732–739.

    Article  PubMed  CAS  Google Scholar 

  31. Zhang, J., L. Chen, T. Su, F. Cao, X. Meng, L. Pei, J. Shi, H.L. Pan, and M. Li. 2010. Electroacupuncture increases CB2 receptor expression on keratinocytes and infiltrating inflammatory cells in inflamed skin tissues of rats. The Journal of Pain 11: 1250–1258.

    Article  PubMed  CAS  Google Scholar 

  32. Gao, F., H.C. Xiang, H.P. Li, M. Jia, X.L. Pan, H.L. Pan, and M. Li. 2018. Electroacupuncture inhibits NLRP3 inflammasome activation through CB2 receptors in inflammatory pain. Brain, Behavior, and Immunity 67: 91–100.

    Article  PubMed  CAS  Google Scholar 

  33. Su, T.F., Y.Q. Zhao, L.H. Zhang, M. Peng, C.H. Wu, L. Pei, B. Tian, J. Zhang, J. Shi, H.L. Pan, and M. Li. 2012. Electroacupuncture reduces the expression of proinflammatory cytokines in inflamed skin tissues through activation of cannabinoid CB2 receptors. European Journal of Pain 16: 624–635.

    Article  PubMed  CAS  Google Scholar 

Download references

Funding

This study was supported by the grants from the Health and Family Planning Commission of Hubei Province (No. 2013Z-Z01) and the Natural Science Foundation of Hubei Province of China (No. 2015CFA096).

Author information

Authors and Affiliations

Authors

Contributions

Tao Chen, Yong Xiong, Nina Yin, and Zebin Chen designed the study; Tao Chen and Yong Xiong performed the experiments and wrote the manuscript; Man Long and Dan Zheng contributed to the rat model and EA pretreatment; Hui Ke helped analyze the data; and Jun Xie and Zebin Chen revised the manuscript.

Corresponding authors

Correspondence to Nina Yin or Zebin Chen.

Ethics declarations

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, T., Xiong, Y., Long, M. et al. Electro-acupuncture Pretreatment at Zusanli (ST36) Acupoint Attenuates Lipopolysaccharide-Induced Inflammation in Rats by Inhibiting Ca2+ Influx Associated with Cannabinoid CB2 Receptors. Inflammation 42, 211–220 (2019). https://doi.org/10.1007/s10753-018-0885-5

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-018-0885-5

KEY WORDS

Navigation