Effects of cysteinyl leukotriene receptors on phagocytosis of mouse microglial cells / 浙江大学学报·医学版

OBJECTIVE@#: To determine the effects of cysteinyl leukotriene receptors (CysLTR and CysLTR) on phagocytosis of mouse BV2 microglial cells.@*METHODS@#: BV2 cells were stimulated with microglial activators lipopolysaccharide (LPS) or CysLT receptor agonists LTD. The phagocytosis of BV2 cells was observed by immunofluorescence analysis and flow cytometry. The intracellular distributions of CysLTR and CysLTR in BV2 cells were examined with immunofluorescence staining.@*RESULTS@#: Both LPS and LTD could significantly enhance the phagocytosis of BV2 cells, and such effect could be inhibited by CysLTR selective antagonist Montelukast and CysLTR selective antagonist HAMI 3379. The activation of BV2 cells induced by LTD or LPS resulted in changes in intracellular distributions of CysLTR and CysLTR. CysLTR and CysLTR was co-localization with a similar distribution.@*CONCLUSIONS@#: CysLTR and CysLTR regulate the phagocytosis of mouse BV2 microglial cells with a synergistic effect.

Expression and distribution of cysteinyl leukotriene receptors CysLT1R and CysLT2R, and GPR17 in brain of Parkinson disease model mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To examine the spatiotemporal profiles and localization of CysLT1R, CysLT2R and GPR17 in mice with 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson disease (PD).</p><p><b>METHODS</b>PD model was induced by subcutaneous injection of MPTP (25 mg/kg) for 5 d in adult male C57BL/6 mice. At d10 after MPTP injection, the expression and cellular localization of CysLT1R, CysLT2R and GPR17 in the substantia nigra were detected by immunohistochemistry and immunofluorescence.</p><p><b>RESULTS</b>CysLT1R, CysLT22 and GPR17 were normally localized in TH-positive dopaminergic neurons and microglia, while CysLT2R was also expressed in astrocytes. In dopaminergic neurons, approximately 91% co-expressed GPR17, 77% co-expressed CysLT1R and 52% co-expressed CysLT2R. Compared with the control group, TH-positive cells in the substantia nigra were significantly reduced in PD mice. CysLT1R, CysLT2R and GPR17-positive cells were significantly reduced; and CysLT1R, CysLT2R, GPR17-positive dopaminergic neurons were also significantly reduced in the PD group. In the striatum, both CysLT1R and GPR17 were normally expressed in neurons; whereas CysLT2R was expressed in astrocytes. In PD striatum, CysLT1R and GPR17-positive cells were decreased, but CysLT2R expression was significantly increased which mainly expressed in the proliferating astrocytes.</p><p><b>CONCLUSION</b>CysLT1R, CysLT2R and GPR17 may be involved in the MPTP-induced PD damage in mice.</p>

Leukotriene D4 activates BV2 microglia in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of CysLT receptor agonist leukotriene D4(LTD4) and antagonists on activation of microglia BV2 cells.</p><p><b>METHODS</b>The expression of CysLT1 and CysLT2 protein was determined by Western blotting and immunostaining in microglia BV2 cells. BV2 cells were pretreated with or without CysLT1 receptor selective antagonist montelukast, CysLT2 receptor selective antagonist HAMI 3379, or CysLT1/CysLT2 receptor dual antagonist BAY u9773 for 30 min, then the cells were treated with LTD4 for 24 h. Cell viability was detected by MTT reduction assay. Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR, respectively.</p><p><b>RESULTS</b>In BV2 cells, LTD4 did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner. LTD4 at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression (P<0.01). HAMI 3379 and BAY u9773 (100 nmol/L) further increased LTD4-induced phagocytosis; BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression, while HAMI 3379 had no effect on that.</p><p><b>CONCLUSION</b>LTD4 activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT1 receptor, LTD4 induces phagocytosis which might be negatively regulated by CysLT2 receptor in BV2 cells.</p>

Effect of Shenmai Injection on angiogenesis in tumor tissue / 中成药

Objective: To investigate the effect of Shenmai Injection (Radix Ginseng Rubra, Radix Ophiopogonis) on angiogenesis and the expression of proliferating cell nuclear antigen (PCNA) in tumor tissue, and the mechanism of it in the treatment of tumor. Methods: The mouse tumor model was used to investigate the effect of Shenmai Injection on tumor growth on the whole. The expression of von Willebrand factor (vWF) and PCNA in tumor tissue was studied by means of immunohistochemistry. Results: Shenmai inhibited the tumor growth and reduced microvessel density and the expression of PCNA in tumor tissue. Conclusion:Antiangiogenesis is one of the mechanisms of Shenmai Injection in treatment of tumor.