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  

Effects of brain-derived neurotrophic factor on colon smooth muscle cells and the mechnism in mice

作　　者：曹静[1] 陈飞雪[1] 王腾飞[1] 赵宏宇[1] 赵栋燕 左秀丽[1] 李延青[1] 

机构地区：[1]山东大学齐鲁医院消化内科,山东济南250012

出　　处：《山东大学学报（医学版）》2015年第12期1-6,共6页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81170352)

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。Objective To investigate the effects of brain-derived neurotrophic factor（ BDNF） on the intracellular Ca2 ＋concentration（ [Ca2 ＋]i） alterations and smooth muscle α-actin（ α-SMA） expression of smooth muscle cells（ SMCs）in mice. Methods The α-SMA expression of colonic SMCs in the BDNF＋ /-mice was measured by Western blotting,and was compared with that in BDNF＋ / ＋mice. The expression of tropomyosin-related kinase B（ Trk B） receptor was identified in the primary colonic SMCs of the mice by immunofluorescence staining. After administration of BDNF and Trk B receptor antagonists（ K252a）,the expressions of α-SMA and Trk B-PLC-Ca2 ＋pathway were measured by Western blotting. The alteration of[Ca2 ＋]iwas measured by[Ca2 ＋]iimaging. Results The expression of α-SMA was obviously decreased in BDNF＋ /-mice compared with that in BDNF＋ / ＋mice. The Trk B receptor was identified in the SMCs. After administration of BDNF,the expressions of α-SMA,Trk B-PLC-Ca2 ＋signal pathway and[Ca2 ＋]i increased. K252 a could reverse those changes. Conclusion BDNF might induce the alterations of[Ca2 ＋]iand α-SMA expression of SMC by Trk B-PLC-Ca2 ＋signal pathway,which might be the mechanism to affect the gut motility.

关 键 词：脑源性神经营养因子 平滑肌细胞 TRK B-PLC-Ca2+信号通路 肠道动力 

分 类 号：R574[医药卫生—消化系统]