基于SDF-1/CXCR4轴探讨雷公藤多糖诱导胰腺干细胞分化的机制研究  

Exploring Mechanism of Tripterygium Wilfordii Polysaccharides Inducing Pancreatic Stem Cell Differentiation Based on SDF-1/CXCR4 Axis

作　　者：贾微[1] 宁志莹 岑妍慧[1] 刘鑫 杨瑞[1] 李博佳 李森 张宸康 JIA Wei;NING Zhiying;CEN Yanhui;LIU Xin;YANG Rui;LI Bojia;LI Sen;ZHANG Chenkang(Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530023

出　　处：《中华中医药学刊》2025年第2期25-28,I0003-I0006,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82260861);广西一流学科建设开放课题(2019XK137);广西一流学科建设项目重点课题(2018XK023,2018XK024);广西高校中青年教师科研基础能力提升项目(2021KY0291);广西中医药大学桂派杏林拔尖人才项目(2022C002);广西中医药大学桂派中医药传承创新团队资助项目(2022B004);广西中医药大学校级重点硕士研究生科研创新项目(YCSZ2022003);广西中医药大学大学生创新创业训练计划项目(202210600017,202010600018);广西中医药大学校级科研项目(2019MS001)。

摘　　要：目的探究雷公藤多糖通过基质细胞衍生因子-1(SDF-1)/CXC型趋化因子受体4(CXCR4)轴诱导胰腺干细胞分化的机制。方法取新生第4天昆明小鼠的胰腺组织制备原代胰腺干细胞,使用免疫组化法鉴定胰腺干细胞;使用双硫腙(DTZ)染色法鉴定胰岛β细胞;使用倒置荧光显微镜对各培养时间点细胞进行拍照记录;使用酶联免疫吸附(ELISA)法检测细胞各给药时间点SDF-1释放量;使用实时荧光定量PCR(qPCR)技术检测各组细胞SDF-1和CXCR4的相对mRNA表达量;使用蛋白免疫印迹试验(WB)检测各组细胞SDF-1和CXCR4蛋白表达量。结果使用免疫组化法鉴定所取细胞Nestin阳性、呈棕黄色;双硫腙(DTZ)染色呈阳性,且给药组细胞阳性染色细胞数量明显高于对照组细胞(P<0.05);对照组细胞SDF-1释放量相对持平,实验组细胞SDF-1释放量随着时间的迁移逐步上升,在第26天达到峰值,随后SDF-1释放量随着时间的迁移逐步下降;qPCR技术检测显示实验组细胞SDF-1和CXCR4的相对mRNA表达量显著高于对照组细胞(P<0.05);WB检测结果显示实验组细胞SDF-1和CXCR4蛋白表达量显著高于对照组细胞(P<0.05)。结论雷公藤多糖可以通过SDF-1/CXCR4轴诱导胰腺干细胞分化。Objective To investigate the mechanism of Tripterygium wilfordii polysaccharides inducing pancreatic stem cell differentiation through stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor-4(CXCR4)axis.Methods Primary pancreatic stem cells were prepared from pancreatic tissue from Kunming mice on day 4 of neonatal birth.The immunohistochemistry method was used to identify the pancreatic stem cells.Isletβcells were identified by dithiazone(DTZ)staining.An inverted fluorescence microscope was used to photograph and record the cells at various cultivation time points.The release of SDF-1 from cells were detected at various administration time points by using enzyme-linked immunosorbent assay(ELISA).The relative mRNA expression levels of SDF-1 and CXCR4 in each group were detected by real-time fluorescence quantitative PCR(qPCR)technology.The expression levels of SDF-1 and CXCR4 proteins in each group were detected by Western blot assay(WB).Results The cells obtained was Nestin positive and Brownish-yellow were identified by immunohistochemistry method.DTZ staining was positive,and the number of positive stained cells in the treatment group was significantly higher than that in the control group(P<0.05).The release of SDF-1 from the control group cells remained relatively stable,while the release of SDF-1 from the experimental group cells gradually increased over time and reached its peak on the 26th day.Subsequently,the release of SDF-1 gradually decreased over time.The relative mRNA expression levels of SDF-1 and CXCR4 in the experimental group cells were significantly higher than those in the control group cells detected by qPCR technology(P<0.05).The expression levels of SDF-1 and CXCR4 proteins in the experimental group cells were significantly higher than those in the control group cells by WB(P<0.05).Conclusion Tripterygium wilfordii polysaccharides can induce pancreatic stem cell differentiation through the SDF-1/CXCR4 axis.

关 键 词：雷公藤多糖 胰腺干细胞 基质细胞衍生因子-1 CXC型趋化因子受体4 

分 类 号：R285.5[医药卫生—中药学]