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

Effect of high-mobility group box 1 on the proliferation of primary neural stem cells in vitro

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-mobilitygroupbox1，HMGBl）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-JunN-terminalkinase，JNK）高选择性抑制剂SP600125探讨其作用机制。结果显示，1和10ng／mLHMGB1分别培养48h$172h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俨〈0．05），而其余浓度HMGBl培养基（0．01、0．1、100ng／mL）与对照组比较无明显差别（P〉0．05）。当HMGB1浓度为10ng／mL时，在神经干细胞增殖的同时P—JNK水平明显升高俨〈0．01）；而10μmol／LSP600125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（1～10ng／mL）HMGB1能够促进神经干细胞增殖，其机制可能是通过促进．INK的磷酸化发挥促增殖作用的。

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 HMGB 1 concentration groups （0.01, 0.1, 100 ng/mL） showed no significant difference, compared with control group （P 〉 0.05）. HMGB1 at l0 ng/mL significantly increased the NSCs proliferation accompanied by the rising ofphosphorylated JNK levels （P 〈 0.01）, and 10 μmol/L SP600125 prevented these effects in HMGBl-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.