| 115 | 0 | 62 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探讨竹节参总皂苷(SPJ)对全氟辛烷磺酸盐(PFOS)的神经毒性以及PC12细胞自噬的干预作用。方法 将PC12细胞暴露于100、200、300、400、500μmol/L的PFOS,利用甲基噻唑基四唑(MTT)法检测细胞活力；Western blot法检测自噬相关蛋白的表达变化。同时采用5、25、50μg/mL SPJ预保护PC12细胞后，用500μmol/L PFOS和不同浓度SPJ共孵育，MTT检测细胞活力；Hoechst染色检测细胞凋亡；2',7'-二氯二氢荧光素二乙酸酯(DCFH-DA)探针标记检测细胞活性氧水平；Western blot法检测自噬相关蛋白LC3、p62蛋白表达变化。结果 不同浓度PFOS暴露后，与对照组比较，100、200、300、400和500μmol/L PFOS显著降低PC12细胞活力(P<0.01),500μmol/L PFOS组LC3蛋白表达显著上升(P<0.05),400和500μmol/L PFOS暴露组p62蛋白表达显著上升(P<0.05),不同浓度PFOS组Beclin1、Atg5、Atg12蛋白表达水平无明显变化；给予不同浓度的SPJ干预后，与500μmol/L PFOS组比较，50μg/mL SPJ组细胞凋亡数量减少，活性氧水平降低(P<0.01),25和50μg/mL SPJ表现为细胞活力明显增高(P<0.01),LC3和p62蛋白表达明显降低(P<0.05)。结论 竹节参总皂苷可通过调节自噬途径减轻PFOS暴露所致PC12细胞损伤。
Abstract：Objective To investigate the neurotoxic effect of Perfluorooctane Sulfonates(PFOS) and the intervention effects of Saponins from Panax japonicus(SPJ) on PC12 cell.Methods The PC12 cells were exposed to 100,200,300,400,500 μmol/L PFOS,cell viability was detected by Methyl Thiazolyl Tetrazolium(MTT).The levels of autophagy-related proteins were assessed using Western blot.PC12 cells were pretreated with different concentrations of SPJ(5,25,50μg/mL),then incubated with 500 μmol/L PFOS and various concentrations of SPJ.The cell viability was detected by MTT.The cell apoptosis was edtected by Hoechst staining.The cellular reactive oxygen species levels were measured by 2',7'-Dichlorodihydrofluorescein diacetate(DCFH-DA) probe labeling.Autophagy-related proteins such as LC3 and p62 were detected through Western blot.Results After exposure to different concentrations of PFOS,the viability of PC12 cells in 100,200,300,400 and 500 μmol/L PFOS groups were significantly decreased(P<0.01),when compared to control cells.The expression of LC3 protein in 500 μmol/L PFOS exposure group was significantly increased(P<0.05),as well as the expression of p62 protein in 400 and 500μmol/L PFOS exposure groups(P<0.05).There were no significant changes in Beclin1,Atg5 and Atg12 protein expression in different concentration PFOS exposure groups.Compared with 500μmol/L PFOS exposure group, the number of apoptotic cells and the levels of reactive oxygen species in the 50μg/mL SPJ group were significantly decreased(P<0.01),Meanwhile, the cell viability was significantly increased(P<0.01),and the expressions of LC3 and p62 proteins were significantly decreased in 25 and 50μg/mL SPJ groups(P<0.05).Conclusion SPJ can reduce the damage of PC12 cells induced by PFOS exposure by regulating autophagy pathway.
[1]HAO Y,CHEN X,WANG K,et al.Fabrication of tailored imprinted layer and cladded layer on magnetic nanomaterials with enhanced specificity for recognition of PFOS[J].Anal Chim Acta,2025,1349:343799
[2]WANG M,LI Q,HOU M,et al.Inactivation of common airborne antigens by perfluoroalkyl chemicals modulates early life allergic asthma[J].Proc Natl Acad Sci U S A,2021,118(24):57
[3]YAO X,SHA S,WANG Y,et al.Perfluorooctane sulfonate induces autophagy-dependent apoptosis through spinster 1-mediated lysosomal-mitochondrial axis and impaired mitophagy[J].Toxicol Sci,2016,153(1):198
[4]国家药典委员会.中国药典[M].北京：中国医药科技出版社，2020:144
[5]WANG T,DI G,YANG L,et al.Saponins from Panax japonicus attenuate D-galactose-induced cognitive impairment through its anti-oxidative and anti-apoptotic effects in rats[J].J Pharm Pharmacol,2015,67(9):1284
[6]REN S,WANG J,DONG Z,et al.Perfluorooctane sulfonate induces ferroptosis-dependent non-alcoholic steatohepatitis via autophagy-MCU-caused mitochondrial calcium overload and MCU-ACSL4 interaction[J].Ecotoxicol Environ Saf,2024,280:116553
[7]SLOTKIN T A,MACKILLOP E A,MELNICK R L,et al.Developmental neurotoxicity of perfluorinated chemicals modeled in vitro[J].Environ Health Perspect,2008,116(6):716
[8]YUAN Z,SHAO X,MIAO Z,et al.Perfluorooctane sulfonate induced neurotoxicity responses associated with neural genes expression,neurotransmitter levels and acetylcholinesterase activity in planarians dugesia japonica[J].Chemosphere,2018,206:150
[9]LI J,FENG R,YANG W,et al.Lysosomal iron accumulation and subsequent lysosomes-mitochondria iron transmission mediate PFOS-induced hepatocyte ferroptosis[J].Ecotoxicol Environ Saf,2024,284:116890
[10]ZHOU A,WANG L,PI X,et al.Effects of perfluorooctane sulfonate (PFOS) on cognitive behavior and autophagy of male mice[J].J Toxicol Sci,2023,48(9):513
[11]ARBOGAST F,SAL-CARRO R,BOUFENGHOUR W,et al.Epidermal maintenance of langerhans cells relies on autophagy-regulated lipid metabolism[J].J Cell Biol,2025,224(2):e202403178
[12]WEI F,WANG Y,YAO J,et al.ZDHHC7-mediated S-palmitoylation of ATG16L1 facilitates LC3 lipidation and autophagosome formation[J].Autophagy,2024,20(12):2719
[13]LIU D,LIU N Y,CHEN L T,et al.Perfluorooctane sulfonate induced toxicity in embryonic stem cell-derived cardiomyocytes via inhibiting autophagy-lysosome pathway[J].Toxicol In Vitro,2020,69:104988
[14]WANG J,HE L,WANG S,et al.Therapeutic effect of the total saponin from Panax Japonicus on experimental autoimmune encephalomyelitis by attenuating inflammation and regulating gut microbiota in mice[J].J Ethnopharmacol,2023,315:116681
[15]QIU L,FENG R,WU Q S,et al.Total saponins from Panax japonicus attenuate acute alcoholic liver oxidative stress and hepatosteatosis by p62-related Nrf2 pathway and AMPK-ACC/PPARalpha axis in vivo and in vitro[J].J Ethnopharmacol,2023,317:116785
[16]PI X J,ZHAO Q Q,WANG J X,et al.Saponins from Panax japonicus attenuate cognitive impairment in ageing rats through regulating microglial polarisation and autophagy[J].Pharm Biol,2021,59(1):1117
[17]DENG L L,WAN J Z,YUAN D,et al.Protective effects of total saponins from Panax japonicus on H2O2 induced injury via mitochondia pathway in SH-SY5Y Cells[J].Zhong Yao Cai,2015,38(8):1690
[18]HUANG H,WANG T,WANG L,et al.Saponins of Panax japonicus ameliorates cardiac aging phenotype in aging rats by enhancing basal autophagy through AMPK/mTOR/ULK1 pathway[J].Exp Gerontol,2023,182:112305
基本信息:
DOI：10.16751/j.cnki.2095-4646.2024122710
中图分类号:R285
引用信息:
[1]刘慧敏,皮雪姣,韩元春,等.竹节参总皂苷调节自噬减轻PFOS暴露所致PC12细胞损伤的研究[J].湖北科技学院学报(医学版),2025,39(05):391-395.DOI:10.16751/j.cnki.2095-4646.2024122710.