中文摘要：

目的：观察益肾化浊方含药脑脊液对神经干细胞（NSCs）增殖与分化的影响,并探讨其相关作用机制。方法：从孕14.5d的昆明小鼠分离培养NSCs,分为空白组,正常组,AG490（JAK/STAT通路抑制剂）组和益肾化浊方组。空白组为培养基本底,其余各组分别予10%的正常脑脊液、AG490和益肾化浊方含药脑脊液。采用CCK-8检测各组NSCs的增殖率,免疫荧光检测其分化情况,Western blot检测Tubulin、GFAP及JAK2/STAT3通路相关蛋白的表达情况。结果：CCK-8检测结果显示：与空白组比较,益肾化浊方组在24h和48h时OD值显著升高（P〈0.05）,而72h时升高无统计学差异。免疫荧光和Western blot检测结果显示：与空白组比较,益肾化浊方组能显著增加Tubulin阳性细胞率及Tubulin蛋白的表达（P〈0.05）,显著降低GFAP阳性细胞率和GFAP蛋白的表达（P〈0.05）,且益肾化浊方组还可显著下调JAK2/STAT3通路中STAT3、p-STAT3、Smad1蛋白的表达（P〈0.05）。结论：益肾化浊方含药脑脊液可促进NSCs的适度增殖,调控NSCs向神经元分化,抑制其向星形胶质细胞分化,该作用可能与抑制JAK2/STAT3通路相关。

英文摘要：

Objective： To explore the effects of cerebrospinal fluid containing Yishen Huazhuo Decoction（YSHZD） on the proliferation and differentiation of neural stem cells（NSCs） via JAK2/STAT3 signaling pathway. Methods： The NSCs were extracted from 14.5 d pregnant Kunming mice, which were cultured in vitro were divided into the blank group, normal group, AG490（JAK/STAT inhibitor） group and the YSHZD group. The blank group was the basic medium, the others were given 10% normal cerebrospinal fluid, AG490, and cerebrospinal fluid containing YSHZD respectively. The proliferation of NSCs was detected by Cell Counting Kit-8（CCK-8）, the differentiation of NSCs was detected by immunofluorescence staining, the protein expression levels of Tubulin, GFAP and others related to JAK2/STAT3 signaling pathway were detected by Western blot. Results： CCK-8 result showed： Compared with the control group, the optical density（OD） of YSHZD group significantly increased at 24 h and 48h（P〈0.05）, but there were no significantly difference between them at 72 h. Immunofluorescence and Western blot showed that compared with the control group, the number of Tubulin-positive cells and the protein expression of Tubulin in QNYZD group significantly increased（P〈0.05）, while the number of GFAP-positive cells and the expression of GFAP reduced（P〈0.05）. Moreover, YSHZD reduced the protein expressions of STAT3, p-STAT3 and Smad1（P〈0.05）. Conclusion： The cerebrospinal fluid containing Yishen Huazhuo Decoction could promote NSCs proliferation, regulate the differentiation of NSCs into neurons, and inhibit the differentiation of astrocytes, which might be related to the inhibition of JAK2/STAT3 signaling pathway.