α-黑素细胞刺激素对原代DMNV细胞钙离子通路及凋亡的影响

Effects of α-MSH on the Ca^(2+) channels of primary DMNV cells and effects of anti-apoptosis on primary DMNV cells after activation of melanocortin receptors

目的:以体外培养的大鼠原代迷走神经运动背核(DMNV)细胞为对象,探讨α-黑素细胞刺激素(α-MSH)对原代DMNV细胞钙离子通路及凋亡的影响。方法:采用体外大鼠原代DMNV细胞培养以及[Ca2+]i测量、实时荧光定量PCR、ELISA细胞凋亡实验等方法进行研究。结果:正常大鼠DMNV组织中存在黑素细胞刺激素受体-4(MC4R)mRNA的表达;NDP-MSH激活MC4R可以促进原代DMNV细胞钙离子内流;NDP-MSH可以降低凝血酶(PAR-1,thrombin)和胰蛋白酶(PAR-2,trypsin)对原代DMNV细胞凋亡。结论:α-MSH可以激发原代DMNV细胞钙离子内流,降低原代DMNV细胞凋亡。

AIM： α-MSH is elevated in patients with inflammatory bowel disease and has been implicated as an inflammatory mediator. The purpose of this study was to investigate effects of α-MSH on the Ca^2＋ channels of primary DMNV cells, the effects of gastrointestinal inflammation on the dorsal motor nucleus of the vagus in rats, as well as the effects of proinflammatory cytokines and α - MSH on neurons from the dorsal motor nucleus of the vagus in vitro. METHODS ： In vitro studies the primary culture of neurons from the dorsal motor nucleus of the vagus was performed. Single - cell cytoplasmic calcium transients were determined using the fluorescence dye fura - 2 - AM. Cell proliferation and apeptosis were measured by enzyme - linked immunosorbent assay. RESULTS ： MC4R mRNA was expressed in the DMNV cells of normal rats. Activation of MC4R promoted the calcium influx of primary DMNV cells. The addition of α-MSH to thrombin or trypsin resulted in significant decreases in apoptosis compared to thrombin or trypsin alone. CONCLUSION： Functionally active α- MSH receptors are linked to Ca^2＋ channels in DMNV neurons. In cultured DMNV cells, α- MSH atten- uates neuronal apoptosis and reverses inhibition of cellular proliferation.