Skip to main content

Advertisement

SpringerLink
Log in

Cytotoxic Effects of Mesaconitine, the Aconitum carmichaelii Debx Bioactive Compound, on HBEC-5i Human Brain Microvascular Endothelial Cells: Role of Ca2+ Signaling-Mediated Pathway

  • Original Article
  • Published:
Neurotoxicity Research Aims and scope Submit manuscript

Abstract

Mesaconitine, one of Aconitum carmichaelii Debx bioactive compounds, was shown to evoke Ca2+ homeostasis and its related physiological effects in endothelial cell types. However, the effect of mesaconitine on Ca2+ signaling and cell viability in human brain microvascular endothelial cells is unclear. This study focused on exploring whether mesaconitine changed cytosolic Ca2+ concentrations ([Ca2+]i), affected cell viability, and established the relationship between Ca2+ signaling and viability in HBEC-5i human brain microvascular endothelial cells. In HBEC-5i cells, cell viability was measured by the cell proliferation reagent (WST-1). [Ca2+]i was measured by the Ca2+-sensitive fluorescent dye fura-2. Mesaconitine (10–100 μM) concentration dependently induced [Ca2+]i rises. Ca2+ removal reduced the signal by approximately 25%. Mesaconitine (40–100 μM) caused cytotoxicity in HBEC-5i cells. This cytotoxic response was significantly reversed by chelation of cytosolic Ca2+ with BAPTA/AM. In Ca2+-containing medium, mesaconitine-induced Ca2+ entry was inhibited by 25% by modulators of store-operated Ca2+ channels and protein kinase C (PKC). Furthermore, mesaconitine also induced Mn2+ influx suggesting of Ca2+ entry. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin abolished mesaconitine-evoked [Ca2+]i rises. Conversely, treatment with mesaconitine abolished thapsigargin-evoked [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 abolished mesaconitine-induced [Ca2+]i rises. In sum, mesaconitine caused cytotoxicity that was triggered by preceding [Ca2+]i rises. Furthermore, mesaconitine induced [Ca2+]i rises by evoking Ca2+ entry via PKC-sensitive store-operated Ca2+ channels and PLC-dependent Ca2+ release from the endoplasmic reticulum. It suggests that Ca2+ signaling have a potential cytotoxic effect on mesaconitine-treated human brain microvascular endothelial cells.

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

Access this article

Log in via an institution

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

  • Al Suleimani YM, Hiley CR (2016) Characterization of calcium signals provoked by lysophosphatidylinositol in human microvascular endothelial cells. Physiol Res 65:53–62

    Article  Google Scholar 

  • Ameri A, Seitz U (1998) Effects of mesaconitine on [3H]noradrenaline uptake and neuronal excitability in rat hippocampus. Exp Brain Res 121:451–456

    Article  CAS  Google Scholar 

  • Baek A, Park EJ, Kim SY, Nam BG, Kim JH, Jun SW, Kim SH, Cho SR (2018) High-frequency repetitive magnetic stimulation enhances the expression of brain-derived neurotrophic factor through activation of Ca2+-calmodulin-dependent protein kinase II-cAMP-response element-binding protein pathway. Front Neurol 9:285

    Article  Google Scholar 

  • Berridge MJ (2016) The inositol trisphosphate/calcium signaling pathway in health and disease. Physiol Rev 96:1261–1296

    Article  CAS  Google Scholar 

  • Berridge MJ (2003) Cardiac calcium signaling. Biochem Soc Trans 31:930–933

    Article  CAS  Google Scholar 

  • Bootman MD (2012) Calcium signaling. Cold Spring Harb Perspect Biol 4:a011171

    Article  Google Scholar 

  • Carrillo C, Giraldo M, Cavia MM, Alonso-Torre SR (2017) Effect of oleic acid on store-operated calcium entry in immune-competent cells. Eur J Nutr 56:1077–1084

    Article  CAS  Google Scholar 

  • Claessens A, Adams Y, Ghumra A, Lindergard G, Buchan CC, Andisi C, Bull PC, Mok S, Gupta AP, Wang CW, Turner L, Arman M, Raza A, Bozdech Z, Rowe JA (2012) A subset of group A-like var genes encodes the malaria parasite ligands for binding to human brain endothelial cells. Proc Natl Acad Sci U S A 109:E1772–E1781

    Article  CAS  Google Scholar 

  • Cui HZ, Kim HY, Kang DG, Lee HS (2013) Ginseng-aconite decoction elicits a positive inotropic effect via the reverse mode Na+/Ca2+ exchanger in beating rabbit atria. J Ethnopharmacol 148:624–631

    Article  CAS  Google Scholar 

  • Dionicio CL, Peña F, Constantino-Jonapa LA, Vazquez C, Yocupicio-Monroy M, Rosales R, Zambrano JL, Ruiz MC, Del Angel RM, Ludert JE (2018) Dengue virus induced changes in Ca2+ homeostasis in human hepatic cells that favor the viral replicative cycle. Virus Res 245:17–28

    Article  CAS  Google Scholar 

  • Dorovini-Zis K, Prameya R, Bowman PD (1991) Culture and characterization of microvascular endothelial cells derived from human brain. Lab Investig 64:425–436

    CAS  PubMed  Google Scholar 

  • Falcón-Moya R, Losada-Ruiz P, Sihra TS, Rodríguez-Moreno A (2018) Cerebellar kainate receptor-mediated facilitation of glutamate release requires Ca2+-calmodulin and PKA. Front Mol Neurosci 11:195

    Article  Google Scholar 

  • Florenzano F, Viscomi MT, Mercaldo V, Longone P, Bernardi G, Bagni C, Molinari M, Carrive P (2006) P2X2R purinergic receptor subunit mRNA and protein are expressed by all hypothalamic hypocretin/orexin neurons. J Comp Neurol 498:58–67

    Article  CAS  Google Scholar 

  • Garg V, Kirichok YY (2019) Patch-clamp analysis of the mitochondrial calcium uniporter. Methods Mol Biol 1925:75–86

    Article  CAS  Google Scholar 

  • Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    Article  CAS  Google Scholar 

  • He LP, Di B, Du YX, Yan F, Su MX, Liu HQ, You LJ (2009) Development and validation of a high-performance liquid chromatography-tandem mass spectrometry method for the rapid simultaneous quantification of aconitine, mesaconitine, and hypaconitine in rat plasma after oral administration of Sini decoction. J Anal Toxicol 33:588–594

    Article  CAS  Google Scholar 

  • Im JY, Joo HJ, Han PL (2011) Rapid disruption of cellular integrity of zinc-treated astroglia is regulated by p38 MAPK and Ca2+-dependent mechanisms. Exp Neurobiol 20:45–53

    Article  Google Scholar 

  • Jaiswal Y, Liang Z, Zhao Z (2016) Botanical drugs in ayurveda and traditional Chinese medicine. J Ethnopharmacol 194:245–259

    Article  Google Scholar 

  • Jiao L, Gan-Schreier H, Tuma-Kellner S, Stremmel W, Chamulitrat W (2015) Sensitization to autoimmune hepatitis in group VIA calcium-independent phospholipase A2-null mice led to duodenal villous atrophy with apoptosis, goblet cell hyperplasia and leaked bile acids. Biochim Biophys Acta 1852:1646–1657

    Article  CAS  Google Scholar 

  • Kim YV, Pearce D, Kim KS (2008) Ca(2+)/calmodulin-dependent invasion of microvascular endothelial cells of human brain by Escherichia coli K1. Cell Tissue Res 332:427–433

    Article  CAS  Google Scholar 

  • Leite PM, Martins MA, Castilho RO (2016) Review on mechanisms and interactions in concomitant use of herbs and warfarin therapy. Biomed Pharmacother 83:14–21

    Article  Google Scholar 

  • Li H, Kan H, He C, Zhang X, Yang Z, Jin J, Zhang P, Ma X (2018) TRPV4 activates cytosolic phospholipase A2 via Ca2+-dependent PKC/ERK1/2 signalling in controlling hypertensive contraction. Clin Exp Pharmacol Physiol 45:908–915

    Article  CAS  Google Scholar 

  • Lorkowska B, Chlopicki S, Marcinkiewicz E, Gryglewski RJ (2004) Statins rise cytoplasmic calcium level [Ca2+]i in cultured endothelial cells. Pol J Pharmacol 56:313–318

    CAS  PubMed  Google Scholar 

  • Lu G, Dong Z, Wang Q, Qian G, Huang W, Jiang Z, Leung KS, Zhao Z (2010) Toxicity assessment of nine types of decoction pieces from the daughter root of Aconitum carmichaeli (Fuzi) based on the chemical analysis of their diester diterpenoid alkaloids. Planta Med 76:825–830

    Article  CAS  Google Scholar 

  • Merritt JE, Jacob R, Hallam TJ (1989) Use of manganese to discriminate between calcium influx and mobilization from internal stores in stimulated human neutrophils. J Biol Chem 264:1522–1527

    Article  CAS  Google Scholar 

  • Morita K, Sakakibara A, Kitayama S, Kumagai K, Tanne K, Dohi T (2002) Pituitary adenylate cyclase-activating polypeptide induces a sustained increase in intracellular free Ca2+ concentration and catechol amine release by activating Ca2+ influx via receptor-stimulated Ca2+ entry, independent of store-operated Ca2+ channels, and voltage-dependent Ca2+ channels in bovine adrenal medullary chromaffin cells. J Pharmacol Exp Ther 302:972–982

    Article  CAS  Google Scholar 

  • Murayama M, Hikino H (1985) Effect of cyclic AMP on mesaconitine-induced analgesia in mice. Eur J Pharmacol 108:19–23

    Article  CAS  Google Scholar 

  • Ogura J, Mitamura M, Someya A, Shimamura K, Takayama H, Aimi N, Horie S, Murayama T (2004) Mesaconitine-induced relaxation in rat aorta: role of Na+/Ca2+ exchangers in endothelial cells. Eur J Pharmacol 483:139–146

    Article  CAS  Google Scholar 

  • Putney JW (2018) Forms and functions of store-operated calcium entry mediators, STIM and Orai. Adv Biol Regul 68:88–96

    Article  CAS  Google Scholar 

  • Sanders K, Moran Z, Shi Z, Paul R, Greenlee H (2016) Natural products for cancer prevention: clinical update 2016. Semin Oncol Nurs 32:215–240

    Article  Google Scholar 

  • Schmitz-Peiffer C (2018) Anarchy in the UPR: a Ca2+-insensitive PKC inhibits SERCA activity to promote ER stress. Biosci Rep 38:BSR20170966

    Article  CAS  Google Scholar 

  • Tan Y, Liu X, Lu C, He X, Li J, Xiao C, Jiang M, Yang J, Zhou K, Zhang Z, Zhang W, Lu A (2014) Metabolic profiling reveals therapeutic biomarkers of processed Aconitum carmichaeli Debx in treating hydrocortisone induced kidney-yang deficiency syndrome rats. J Ethnopharmacol 152:585–593

    Article  CAS  Google Scholar 

  • Thastrup O, Cullen PJ, Drøbak BK, Hanley MR, Dawson AP (1990) Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+-ATPase. Proc Natl Acad Sci U S A 87:2466–2470

    Article  CAS  Google Scholar 

  • Tong P, Xu S, Cao G, Jin W, Guo Y, Cheng Y, Jin H, Shan L, Xiao L (2014) Chondroprotective activity of a detoxicated traditional Chinese medicine (Fuzi) of Aconitum carmichaeli Debx against severe-stage osteoarthritis model induced by mono-iodoacetate. J Ethnopharmacol 151:740–744

    Article  CAS  Google Scholar 

  • Tsien RY (1980) New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry 19:2396–2404

    Article  CAS  Google Scholar 

  • Wei H, Wu H, Yu W, Yan X, Zhang X (2015) Shenfu decoction as adjuvant therapy for improving quality of life and hepatic dysfunction in patients with symptomatic chronic heart failure. J Ethnopharmacol 169:347–355

    Article  Google Scholar 

  • Xu XL, Yang LJ, Jiang JG (2016) Renal toxic ingredients and their toxicology from traditional Chinese medicine. Expert Opin Drug Metab Toxicol 12:149–159

    Article  CAS  Google Scholar 

  • Zhang F, Tang MH, Chen LJ, Li R, Wang XH, Duan JG, Zhao X, Wei YQ (2008) Simultaneous quantitation of aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine and benzoylhypaconine in human plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetics evaluation of “SHEN-FU” injectable powder. J Chromatogr B Anal Technol Biomed Life Sci 873:173–179

    Article  CAS  Google Scholar 

  • Zhang M, Peng CS, Li XB (2015) In vivo and in vitro metabolites from the main diester and monoester diterpenoid alkaloids in a traditional Chinese herb, the Aconitum species. Evid Based Complement Alternat Med 2015:252434

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by the Department of Pharmacy and Master Program, College of Pharmacy and Health Care, Tajen University, Pingtung County 90741, Taiwan.

Author information

Authors and Affiliations

  1. Department of Surgery, Kaohsiung Veterans General Hospital, Kaohsiung, 81362, Taiwan

    Shu-Shong Hsu

  2. Department of Surgery, National Defense Medical Center, Taipei, 11490, Taiwan

    Shu-Shong Hsu

  3. College of Health and Nursing, Meiho University, Neipu, Pingtung, 91202, Taiwan

    Shu-Shong Hsu

  4. Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, 81362, Taiwan

    Wei-Zhe Liang

  5. Department of Pharmacy and Master Program, College of Pharmacy and Health Care, Tajen University, Pingtung County, 90741, Taiwan

    Wei-Zhe Liang

Authors
  1. Shu-Shong Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wei-Zhe Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wei-Zhe Liang.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hsu, SS., Liang, WZ. Cytotoxic Effects of Mesaconitine, the Aconitum carmichaelii Debx Bioactive Compound, on HBEC-5i Human Brain Microvascular Endothelial Cells: Role of Ca2+ Signaling-Mediated Pathway. Neurotox Res 39, 256–265 (2021). https://doi.org/10.1007/s12640-020-00249-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12640-020-00249-2

Keywords

Search

Navigation