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目的:研究炎性肠病大鼠模型的迷走神经背侧运动核(DMNV)蛋白酶激活受体(PAR-1,PAR-2)存在的情况,并阐明该受体激活的机制。方法:制备20只炎性肠病的大鼠模型中取DMNV组织检测PAR-1和PAR-2;培养新出生的大鼠DMNV原代细胞,利用钙离子荧光探针Fura-2-AM检测PAR-1和PAR-2及各种影响因素对细胞钙内流的影响。结果:凝血酶和其类似物PARP-1可以分别激活PAR-1出现最大钙离子内流223.3%±23.5%和145.6%±17.2%;胰蛋白酶和其类似物PARP-2分别激活PAR-2出现最大钙离子内流242.7%±28.7%和236.7%±19.8%。使用1μmol/L磷脂酶C抑制剂U73312可以降低PAR-1激活的细胞钙离子内流140.1%±16.5%到20.7%±2.5%;降低PAR-2激活的钙离子内流225.4%±20.5%到45.4%±5.6%。钙离子抑制剂2APB可以降低PAR-1和PAR-2激活钙离子内流149.7%±13.4%和195.1%±21.5%分别到63.2%±4.3%和75.3%±13.5%。结论:在DMNV中存在PAR-1和PAR-2,它们激活后通过磷脂酶C激活和1,4,5-三磷酸肌醇信号通路参与进行调解钙离子内流。
Objective: To investigate the existence of PAR-1 and PAR-2 in the rat model of inflammatory bowel disease and to elucidate the mechanism of its activation. Methods: DMNV tissue was used to detect PAR-1 and PAR-2 in a rat model of 20 inflammatory bowel disease. Primary neonatal rat DMNV cells were cultured and detected by Fura-2-AM, a calcium ion fluorescent probe -1 and PAR-2 and various influencing factors on the intracellular calcium influx. Results: Thrombin and its analogues PARP-1 could activate PAR-1 and Ca (superscript 2 +) respectively, and the highest Ca (superscript 2 +) influx was 223.3% ± 23.5% and 145.6% ± 17.2% respectively; PAR-2 was activated by trypsin and its analog PARP- The maximum calcium influx was 242.7% ± 28.7% and 236.7% ± 19.8%. The use of 1μmol / L phospholipase C inhibitor U73312 decreased the influx of intracellular Ca2 + from 140.1% ± 16.5% to 20.7% ± 2.5% in PAR-1-activated cells and 225.4% ± 20.5% to PAR-2- 45.4% ± 5.6%. Calcium ion inhibitor 2 APB reduced the influx of PAR-1 and PAR-2 by 149.7% ± 13.4% and 195.1% ± 21.5% to 63.2% ± 4.3% and 75.3% ± 13.5%, respectively. CONCLUSIONS: PAR-1 and PAR-2 are present in DMNV and are activated by phospholipase C activation and the inositol 1,4,5-triphosphate signaling pathway to mediate calcium influx.