脑源性神经营养因子对小鼠结肠平滑肌细胞的作用及其机制

发布时间：2018-12-12 16:49

【摘要】：目的探讨脑源性神经营养因子(BDNF)对小鼠结肠平滑肌细胞(SMC)钙离子浓度及α-平滑肌激动蛋白(α-SMA)的影响及其相关调节机制。方法 Western blotting法检测BDNF基因敲除(BDNF~(+/-))小鼠与正常野生型(BDNF~(+/+))小鼠α-SMA表达水平的差异。培养原代小鼠结肠SMC,免疫荧光法检测SMC中酪氨酸激酶B(Trk B)受体的表达。同时以BDNF、TrkB受体阻滞剂(K252a)干预SMC,Western blotting法检测α-SMA和Trk BPLC-Ca~(2+)信号通路蛋白表达水平的变化,钙离子成像法检测SMC内钙离子浓度的变化。结果与BDNF+/+小鼠相比,BDNF~(+/-)小鼠结肠α-SMA表达水平明显降低。SMC表达Trk B受体,在BDNF作用下,SMC中α-SMA、Trk B-PLC-Ca~(2+)信号通路蛋白表达量和细胞内钙离子浓度增加,且加入K252a可阻断以上变化。结论 BDNF可能通过Trk B-PLC-Ca~(2+)信号通路作用于SMC,影响细胞内钙离子浓度及α-SMA的表达水平,进而影响肠道动力。
[Abstract]:Objective to investigate the effects of brain-derived neurotrophic factor (BDNF) on the calcium concentration of (SMC) and 伪 -smooth muscle activin (伪 -SMA) in colonic smooth muscle cells of mice and its related regulatory mechanisms. Methods the expression of 伪-SMA in BDNF knockout (BDNF~ (/ -) mice and normal wild type (BDNF~ (/) mice was detected by Western blotting method. The expression of tyrosine kinase B (Trk B) receptor in primary mouse colon was detected by SMC, immunofluorescence assay. At the same time, BDNF,TrkB receptor blocker (K252a) was used to detect the expression of 伪 SMA and Trk BPLC-Ca~ (2) signaling pathway protein by SMC,Western blotting method, and calcium imaging was used to detect the change of Ca 2 + concentration in SMC. Results compared with BDNF / mice, the expression of 伪-SMA in colon of BDNF~ (/ -) mice was significantly lower than that of BDNF / mice. SMC expressed Trk B receptor, and 伪-SMA, in SMC was induced by BDNF. Trk B-PLC-Ca2 signaling pathway protein expression and intracellular calcium concentration increased, and K252a could block these changes. Conclusion BDNF may affect the intracellular Ca ~ (2 +) concentration and 伪-SMA expression through Trk B-PLC-Ca2 signaling pathway, and then affect intestinal motility.
【作者单位】：山东大学齐鲁医院消化内科;
【基金】：国家自然科学基金(81170352)
【分类号】：R574

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