Yan Long

Publications (2)4.54 Total impact

• Article: Functional comparison of the reverse mode of Na(+)/Ca(2+) exchangers NCX1.1 and NCX1.5 expressed in CHO cells.

  Yan Long, Wei-Ping Wang, Hui Yuan, Shi-Ping Ma, Nan Feng, Ling Wang, Xiao-Liang Wang
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  ABSTRACT: Aim:To investigate the reverse mode function of Na(+)/Ca(2+) exchangers NCX1.1 and NCX1.5 expressed in CHO cells as well as their modulations by PKC and PKA.Methods:CHO-K1 cells were transfected with pcDNA3.1 (+) plasmid carrying cDNA of rat cardiac NCX1.1 and brain NCX1.5. The expression of NCX1.1 and NCX1.5 was examined using Western blot analysis. The intracellular Ca(2+) level ([Ca(2+)]i) was measured using Ca(2+) imaging. Whole-cell NCX currents were recorded using patch-clamp technique. Reverse mode NCX activity was elicited by perfusion with Na(+)-free medium. Ca(2+) paradox was induced by Ca(2+)-free EBSS medium, followed by Ca(2+)-containing solution (1.8 or 3.8 mmol/L CaCl2).Results:The protein levels of NCX1.1 and NCX1.5 expressed in CHO cells had no significant difference. The reverse modes of NCX1.1 and NCX1.5 in CHO cells exhibited a transient increase of [Ca(2+)]i, which was followed by a Ca(2+) level plateau at higher external Ca(2+) concentrations. In contrast, the wild type CHO cells showed a steady increase of [Ca(2+)]i at higher external Ca(2+) concentrations. The PKC activator PMA (0.3-10 μmol/L) and PKA activator 8-Br-cAMP (10-100 μmol/L) significantly enhanced the reverse mode activity of NCX1.1 and NCX1.5 in CHO cells. NCX1.1 was 2.4-fold more sensitive to PKC activation than NCX1.5, whereas the sensitivity of the two NCX isoforms to PKA activation had no difference. Both PKC- and PKA-enhanced NCX reverse mode activities in CHO cells were suppressed by NCX inhibitor KB-R7943 (30 μmol/L).Conclusion:Both NCX1.1 and NCX1.5 are functional in regulating and maintaining stable [Ca(2+)]i in CHO cells and differentially regulated by PKA and PKC. The two NCX isoforms might be useful drug targets for heart and brain protection.
  Acta Pharmacologica Sinica 04/2013; · 1.95 Impact Factor
• Article: Potassium 2-(1-hydroxypentyl)-benzoate attenuated hydrogen peroxide-induced apoptosis in neuroblastoma SK-N-SH cells.

  Yanli Hu, Ying Peng, Yan Long, Shaofeng Xu, Nan Feng, Ling Wang, Xiaoliang Wang
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  ABSTRACT: Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB) has been shown to have potent neuroprotective effects, such as reducing the infarct volume and improving neurobehavioral deficits in the transient focal cerebral ischemic rat model. The present study is to evaluate the neuroprotective effect of dl-PHPB on hydrogen peroxide (H(2)O(2))-induced apoptosis and the possible mechanism in the human neuroblastoma SK-N-SH cells. Our results showed that dl-PHPB significantly attenuated H(2)O(2)-induced cell death, and reduced neuronal apoptosis. Dl-PHPB partially reversed the decrease of B-cell CLL/lymphoma 2 (Bcl-2) protein level induced by H(2)O(2). Furthermore, dl-PHPB inhibited the elevation of pro-apoptotic Bcl-2-associated X protein (Bax) and caspase3, and alleviated the down-regulation of protein kinase C alpha (PKCα). The PKC inhibitor, Calphostin C significantly attenuated the protective effects of dl-PHPB. The findings suggest that dl-PHPB may protect neurons against H(2)O(2)-induced apoptosis by modulating apoptosis-related proteins, and PKC signaling pathway may be involved in the neuroprotection of dl-PHPB.
  European journal of pharmacology 04/2012; 680(1-3):49-54. · 2.59 Impact Factor