Mechanism of melatonin through paraventricular nucleus against myocardial ischemia/reperfusion injury
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摘要:
目的 探讨外周输注褪黑素通过下丘脑室旁核（PVN）调节交感神经活性对抗心肌I/R损伤的机制。 方法 腹腔注射褪黑素或者无菌生理盐水，24 h后建立C57BL/6J小鼠心肌I/R损伤模型。超声心动图测定心功能，用Evans blue/TTC双染色法测定再灌注24 h后心肌梗死面积。免疫荧光检测下丘脑室旁核（PVN）IL-10和IL-1β及心肌酪氨酸羟化酶（TH）水平；Western blot检测下丘脑室旁核NF-κB水平；ELISA检测血浆去甲肾上腺素(NE)水平。 结果 外周给予褪黑素可抑制PVN区NF-κB水平（P<0.01），进而降低PVN区IL-1β水平（P<0.01），升高IL-10水平（P<0.01），降低NE及TH活性，缩小心肌梗死面积（P<0.01），改善C57BL/6J小鼠心功能。 结论 外周注射褪黑素，通过调控PVN区炎性细胞因子水平，抑制交感神经活性，改善心肌I/R损伤所致心功能降低。 -
关键词:
- 心肌缺血/再灌注损伤 /
- 褪黑素 /
- 下丘脑室旁核 /
- 交感神经活性 /
- 炎性细胞因子
Abstract:AIM To explore the mechanism of peripheral injection of melatonin in regulating sympathetic activity through hypothalamic paraventricular nucleus (PVN) against myocardial I/R injury. METHODS The myocardial I/R injury model of C57BL/6J MICE was established 24 hours after intraperitoneal injection of melatonin or sterile saline. T cardiac functions were measured by echocardiography and the infarct size was measured by Evans blue/TTC double staining after 24 hours of reperfusion. The levels of Il-10 and il-1β in paraventricular nucleus of hypothalamus were detected by immunofluorescence assay, the levels of NF-κb in paraventricular nucleus of hypothalamus and myocardial Tyrosine 3-monooxygenase were detected by Western blot and the level of (-)-noradrenaline in plasma was detected by ELISA. RESULTS Peripheral administration of melatonin inhibited NF-ΚB level in PVN region, decreased IL-1 β level in PVN region, increased IL-10 level, decreased NE and TH activity, reduced infarct size （all P<0.01） and improved cardiac functions in C57BL/6J mice. CONCLUSION Peripheral injection of melatonin can regulate the level of inflammatory cytokines in PVN, inhibit the activity of sympathetic nervous system and improve the cardiac function decline induced by myocardial I/R injury. -
图 1 腹腔注射Mel对心肌I/R损伤的保护作用
A：m型超声心动图图像；B： m型超声心动图定量分析LVEF和LVFS；C：Evans blue/TTC双染左室横切切片；D：定量分析心肌梗死面积及心肌危险区占左室总面积百分比。1：Sham+V组；2：Sham+Mel组；3：MI/R+V组；4：MI/R+Mel组。n=6。与Sham+V组比较，bP<0.01；与MI/R+V组比较，dP<0.01
图 2 Mel通过抑制PVN区NF-κB水平发挥调控炎性因子的作用
A：Western blot检测PVN组织中NF-κBp65蛋白表达； B：各组中NF-κBp65蛋白密度分析。1：Sham+V组；2：Sham+Mel组；3：MI/R+V组；4：MI/R+Mel组。n=6。与Sham+V组比较，bP<0.01；与MI/R+V组比较，dP<0.01
图 3 Mel调控MI/R损伤后PVN区炎性因子水平
A：免疫荧光检测PVN组织中IL-1β和IL-10表达；B：各组中IL-1β免疫荧光强度的光密度分析；C：各组中IL-10免疫荧光强度的光密度分析。1：Sham+V组；2：Sham+Mel组；3：MI/R+V组；4：MI/R+Mel组。n=6。与Sham+V组比较，bP<0.01；与MI/R+V组比较，dP<0.01
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