文献类型：期刊文献

英文题名：Anti-inflammatory and DNA Repair Effects of Astragaloside IV on PC12 Cells Damaged by Lipopolysaccharide

作者：Li, Hai-long[1,2];Shao, Li-hua[2];Chen, Xi[2];Wang, Meng[2];Qin, Qi-jie[3];Yang, Ya-li[2,4];Zhang, Guang-run[5];Hai, Yang[6];Tian, Yi-hong[4,6]

第一作者：Li, Hai-long;李海龙

通信作者：Tian, YH[1];Hai, Y[2];Tian, YH[2]

机构：[1]Gansu Univ Chinese Med, Affiliated Hosp, Dept Geriatr, Lanzhou 730000, Peoples R China;[2]Key Lab Min Innovat & Transformat Tradit Chinese M, Lanzhou 730000, Peoples R China;[3]First Peoples Hosp Lanzhou, Dept Neurol, Lanzhou 730000, Gansu, Peoples R China;[4]Gansu Univ Chinese Med, Sch Publ Hlth, Lanzhou 730000, Peoples R China;[5]Peoples Hosp Gansu Prov, Lanzhou 730000, Peoples R China;[6]Gansu Univ Chinese Med, Sci Res & Expt Ctr, Lanzhou 730000, Peoples R China

第一机构：甘肃中医药大学第二附属医院

通信机构：[1]corresponding author), Gansu Univ Chinese Med, Sch Publ Hlth, Lanzhou 730000, Peoples R China;[2]corresponding author), Gansu Univ Chinese Med, Sci Res & Expt Ctr, Lanzhou 730000, Peoples R China.|[107359c91a78c5fd2e803]甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）;[10735]甘肃中医药大学;[10735e9d5e7087247e71b]甘肃中医药大学公共卫生学院;

年份：2024

外文期刊名：CURRENT MEDICAL SCIENCE

收录：;Scopus(收录号：2-s2.0-85200577585);WOS:【SCI-EXPANDED(收录号:WOS:001286126800001)】；

基金：The study was supported by grants from Open Project of GansuTraditional Chinese Medicine Research Center (No. zyzx-2020-10),Gansu Province Youth Science and Technology Foundation Program (No. 21JR7RA652), Gansu Province Higher Education Research (No. 2018A-049) and Gansu Province Higher Education Research (No.2021B-163)

语种：英文

外文关键词：PC12 cells; astragaloside IV; inflammation; DNA damage

摘要：Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide (LPS) and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses. Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25, 0.5, 0.75, 1, and 1.25 mg/mL for 24 h. Cell morphology was evaluated, and cell survival rates were calculated. A neurocyte inflammatory model was established with LPS treatment, which reached a 50% cell survival rate. PC12 cells were treated with 0.01, 0.1, 1, 10, or 100 mu mol/L astragaloside IV for 24 h. The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments. NOS activity was detected by colorimetry; the expression levels of ERCC2, XRCC4, XRCC2, TNF-alpha, IL-1 beta, TLR4, NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting. The differentially expressed genes (DEGs) between the groups were screened using a second-generation sequence (fold change>2, P<0.05) with the following KEGG enrichment analysis, RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells. Results The viability of PC12 cells was not altered by treatment with 0.01, 0.1, or 1 mol/L astragaloside IV for 24 h (P>0.05). However, after treatment with 0.5, 0.75, 1, or 1.25 mg/mL LPS for 24 h, the viability steadily decreased (P<0.01). The mRNA and protein expression levels of ERCC2, XRCC4, XRCC2, TNF-alpha, IL-1 beta, TLR4, NOS, and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h (P<0.01); however, these changes were reversed when PC12 cells were pretreated with 0.01, 0.1, or 1 mu mol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h (P<0.05). Second-generation sequencing revealed that 1026 genes were upregulated, while 1287 genes were downregulated. The DEGs were associated with autophagy, TNF-alpha, interleukin-17, MAPK, P53, Toll-like receptor, and NOD-like receptor signaling pathways. Furthermore, PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2, CCL11, CCL7, MMP3, and MMP10, which are associated with the IL-17 pathway. RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results. Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage. astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.