Distinct roles of CysLT1 and CysLT2 receptors in oxygen glucose deprivation-induced PC12 cell death
Section snippets
Materials and methods
Cell culture and receptor gene transfection. PC12 cells were purchased from the Institute of Cell Biology, Chinese Academy of Sciences, Shanghai, China. The cDNA for mouse CysLT1 or CysLT2 receptor (mCysLT1 and mCysLT2, subcloned into pcDNA3.0) was kindly gifted by Professor C.D. Funk (University of Pennsylvania, USA). The pcDNA3.0 null vector was purchased from Invitrogen (Carlsbad, California, USA). The receptor cDNA expressing vectors and the null vector were lineared by PvuI and transfected
Expressions of CysLT1 and CysLT2 receptor mRNAs
The endogenous mRNA expression of rat CysLT2 receptor (rCysLT2) was higher than that of rCysLT1 in normal PC12 cells. Permanent transfection with mCysLT1 and mCysLT2 receptors increased their mRNA expressions (Fig. 1A). The over-expression of CysLT1 receptor did not alter whereas over-expression of CysLT2 receptor significantly increased the cell size (Fig. 1B).
Hoechst 33258 and PI double staining
OGD mainly induced PC12 cell apoptosis. After 6-h OGD, apoptotic cells were less in PC12/mCysLT1 cells (29.0%, P < 0.01) and more in
Discussion
The most important finding in the present study is that over-expression of CysLT1 receptor reduced but over-expression of CysLT2 receptor increased OGD-induced PC12 cell death, indicating the distinct roles of CysLT1 and CysLT2 receptors in ischemic neuronal injury. The distinct roles of the two receptors have also been reported in mouse studies in which bleomycin-induced pulmonary fibrosis is enhanced in CysLT1 receptor-deficient mice but reduced in CysLT2 receptor-deficient mice [17], [18],
Acknowledgments
This study was supported by the National Natural Science Foundation of China (No. 30500613) and the Scientific Foundation of Education Ministry of China (20050335105).
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Cited by (29)
Autocrine activity of cysteinyl leukotrienes in human vascular endothelial cells: Signaling through the CysLT<inf>2</inf> receptor
2015, Prostaglandins and Other Lipid MediatorsCitation Excerpt :Indeed, locally formed cysteinyl-LTs resulting from transcellular biosynthesis have been shown to affect coronary [3–5] and cerebral vessels [6], resulting in myocardial ischemia and significant alteration of vascular permeability. Furthermore, a number of papers have reported the involvement of either CysLT2 [52] or both CysLT1 and CysLT2 receptors [53–55] in the inflammatory process subsequent to a brain vascular insult, such as vascular ischemia or oxygen deprivation. Taking advantage of a novel model of PMNL-perfused isolated rat brain we provide evidence that this alteration of vascular permeability is the result of Cys-LT2R activation, further supporting the notion that this receptor may represent an important target for pharmacological interventions aimed at modulating endothelial dysfunction in CV pathologies.
HAMI 3379, a CysLT<inf>2</inf>R antagonist, dose- and time-dependently attenuates brain injury and inhibits microglial inflammation after focal cerebral ischemia in rats
2015, NeuroscienceCitation Excerpt :CysLT2R knockout or transgenic strategies in mice have demonstrated the involvement of this receptor in inflammatory responses and pulmonary fibrosis (Beller et al., 2004), vascular permeability (Hui et al., 2004; Jiang et al., 2008; Moos et al., 2008; Ni et al., 2014) and myocardial ischemia/reperfusion injury (Jiang et al., 2008). It has been reported that CysLT2R expression is induced in brain trauma and tumors (Hu et al., 2005), and it mediates ischemia-like injury in PC12 cells (Sheng et al., 2006). In the brain of rats with focal cerebral ischemia, CysLT2R expression is increased, and the upregulated receptor is restricted to injured neurons in the ischemic core in the acute phase (∼24 h); however, in the late phase (∼14 days), it is mainly distributed in hypertrophic microglia in the ischemic core and proliferated astrocytes in the boundary zone (Fang et al., 2007; Zhao et al., 2011a).
Intracerebroventricular injection of HAMI 3379, a selective cysteinyl leukotriene receptor 2 antagonist, protects against acute brain injury after focal cerebral ischemia in rats
2012, Brain ResearchCitation Excerpt :It has been reported that CysLT2R inhibits CysLT1R-mediated mast cell proliferation (Jiang et al., 2007) and CysLT1R internalization (Parhamifar et al., 2010) through their interaction. Since both CysLT1R and CysLT2R have similar spatiotemporal patterns of expression and distribution (Fang et al., 2006; Fang et al., 2007; Zhao et al., 2011a), CysLT2R might alter the CysLT1R-mediated attenuation of ischemic injury that is reported in PC12 cells (Sheng et al., 2006). BBB breakdown resulting from vascular damage occurs within 3–5 h and persists for several days is a major ischemic insult (Gotoh et al., 1985).
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These authors contributed equally to this work.