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高迁移率族蛋白B1对体外培养原代神经干细胞增殖能力的影响

李满, 罗勇, 李圆, 孙麟

山西医科大学第二医院神经内科，太原 030001；重庆医科大学附属第一医院神经内科，重庆市神经病学重点实验室，重庆 400016；山西医科大学基础医学院，太原 030001；山西医学科学院山西大医院骨科，太原 030032

摘要

本研究采用CCK-8增殖实验和测量神经球直径的方法探讨不同浓度高迁移率族蛋白B1 (high-mobility group box 1, HMGB1)对体外培养原代大鼠海马神经干细胞增殖能力的影响，同时通过使用c-Jun氨基端激酶(c-Jun N-terminal kinase, JNK)高选择性抑制剂SP600125探讨其作用机制。结果显示，1和10 ng/mL HMGB1分别培养48 h和72 h后，神经干细胞的CCK-8吸光度值和神经球直径与对照组比较有明显增加(P < 0.05)，而其余浓度HMGB1培养基(0.01、0.1、100 ng/mL)与对照组比较无明显差别(P > 0.05)。当HMGB1浓度为10 ng/mL时，在神经干细胞增殖的同时p-JNK水平明显升高(P < 0.01)；而10 μmol/L SP600125明显抑制HMGB1促进神经干细胞增殖的作用，降低p-JNK水平(P < 0.01)。结果表明，低浓度(1~10 ng/mL) HMGB1能够促进神经干细胞增殖，其机制可能是通过促进JNK的磷酸化发挥促增殖作用的。

关键词： 神经干细胞; 增殖; 高迁移率族蛋白B1; CCK-8; 神经球直径; c-Jun氨基端激酶

分类号：Q2-33

[Effect of high-mobility group box 1 on the proliferation of primary neural stem cells in vitro.] [Article in Chinese]

LI Man, LUO Yong, LI Yuan, SUN Lin

Department of Neurology, the Second Affiliated Hospital of Shanxi Medical University, Taiyuan 030001, China； Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Nerology, Chongqing, 400016 , China； Basic Medicine college of Shanxi Medical University, Taiyuan 030001, China； Department of Orthopedics, Shanxi Academy of Medical Sciences, Shanxi Dayi Hospital, Taiyuan 030032, China

Abstract

The cell counting kit-8 (CCK-8) proliferation assay and diameter measure of neurospheres were used to investigate the effect of high-mobility group box 1 (HMGB1) on proliferation of primary rat neural stem cells (NSCs) in vitro, and c-Jun N-terminal protein kinase (JNK) potent inhibitor SP600125 was used to study the mechanism. The results demonstrated that HMGB1 significantly increased CCK-8 absorbance values and neurosphere diameters at concentrations of 1 and 10 ng/mL at 48 h and 72 h (P < 0.05), and the other HMGB1 concentration groups (0.01, 0.1, 100 ng/mL) showed no significant difference, compared with control group (P > 0.05). HMGB1 at 10 ng/mL significantly increased the NSCs proliferation accompanied by the rising of phosphorylated JNK levels (P < 0.01), and 10 μmol/L SP600125 prevented these effects in HMGB1-cultured NSCs (P < 0.01). In conclusion, low concentration of HMGB1 (1-10 ng/mL) can increase NSCs proliferation, which may play a role by promoting JNK phosphorylation.

Key words: neural stem cell; proliferation; high-mobility group box 1; CCK-8; neurosphere diameter; JNK

收稿日期：2013-11-22　　录用日期：2014-01-02

通讯作者：罗勇　　E-mail: luoyong1998@163.com

引用本文：

李满, 罗勇, 李圆, 孙麟. 高迁移率族蛋白B1对体外培养原代神经干细胞增殖能力的影响[J]. 生理学报 2014; 66 (4): 469-475.

LI Man, LUO Yong, LI Yuan, SUN Lin. [Effect of high-mobility group box 1 on the proliferation of primary neural stem cells in vitro.] [Article in Chinese]. Acta Physiol Sin 2014; 66 (4): 469-475 (in Chinese with English abstract).