中医学报2024，Vol.39Issue(8)：1583-1590,8.DOI:10.16368/j.issn.1674-8999.2024.08.262
基于miR-335/ROCK2轴探讨补肾启智方对OGD/R损伤HT22细胞焦亡与应激颗粒的影响
Effects of Bushen Qizhi Formula on OGD/R Injury Induced HT22 Cell Pyroptosis and Stress Granules Based on miR-335/ROCK2 Axis
摘要
Abstract
Objective:To investigate the effect of Bushen Qizhi Formula(BSQZ)on oxygen-glucose deprivation/reoxygenation(OGD/R)injury induced HT22 cell pyroptosis and stress granules(SGs)by regulating microRNA-335(miR-335)targeting Rho-associ-ated protein kinase 2(ROCK2).Methods:HT22 cells in logarithmic growth phase were randomly divided into a blank group,a model group,a BSQZ group,a miR-335 inhibition group,an empty virus control group,and BSQZ+miR-335 inhibition group.Except for the blank group,OGD/R injury models were established in the cells of the other groups.The cell survival rate was detected using CCK-8 method,the expression level of miR-335 mRNA was detected with qRT-PCR,and the expressions of ROCK2,T cell intracellular anti-gen 1(TIA1),nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like pro-tein containing CARD(ASC),c-Caspase-1 and gasdermin D-N(GSDMD-N)protein were detected using Western blot.ELISA was used to detect interleukin-1β(IL-1β)and interleukin-18(IL-18)levels in cell supernatant.Results:Compared with that of the blank group,the cell survival rate and miR-335 mRNA level of the model group were decreased significantly(P<0.05).Com-pared with that of the model group,the cell survival rate and miR-335 mRNA level of the BSQZ group were significantly increased,while that of the miR-335 inhibition group were decreased significantly(P<0.01).Compared with that of the empty virus control group,the cell survival rate and miR-335 mRNA level of the miR-335 inhibition group were decreased significantly(P<0.01).Compared with that of the miR-335 inhibition group,the cell survival rate and miR-335 mRNA level of the BSQZ+miR-335 inhi-bition group were significantly increased(P<0.01).Compared with that of the blank group,the protein expression levels of ROCK2,NLRP3,ASC,c-Caspase-1 and GSDMD-N were significantly increased,while the TIA1 protein expression level of the model group was significantly decreased(P<0.01).Compared with that of the model group,the protein expression levels of ROCK2,NLRP3,ASC,c-Caspase-1 and GSDMD-N were decreased significantly,while the protein expression level of TIA1 of the BSQZ group was signifi-cantly increased,however,the expression trend of the above proteins of the miR-335 inhibition group was opposite(P<0.05).Com-pared with that of the empty virus control group,the protein expression levels of ROCK2,NLRP3,ASC,c-Caspase-1 and GSDMD-N of the miR-335 inhibition group were significantly increased,while the protein expression level of TIA1 was decreased significantly(P<0.05).Compared with that of the miR-335 inhibition group,the expression trend of the above proteins of the BSQZ+miR-335 inhibition group was opposite(P<0.01).Compared with that of the blank group,the levels of IL-1β and IL-18 of the model group were increased significantly(P<0.01).Compared with that of the model group,the levels of IL-1β and IL-18 of the BSQZ group were decreased significantly(P<0.01),which was opposite of the miR-335 inhibition group(P<0.05).Compared with that of the empty virus control group,the levels of IL-1β and IL-18 of the miR-335 inhibition group were increased significantly(P<0.05).Compared with that of the miR-335 inhibition group,the expression trend of the above factors of the BSQZ+miR-335 inhibition group was opposite(P<0.01).Conclusion:Bushen Qizhi Formula may play a neuroprotective role by regulating miR-335/ROCK2 activity to promote stress granule formation,inhibit inflammatory response and NLRP3-mediated pyroptosis.关键词
补肾启智方/卒中后认知障碍/海马/氧糖剥夺/复氧/miR-335/ROCK2/NLRP3炎症小体/应激颗粒/细胞焦亡Key words
Bushen Qizhi Formula/post-stroke cognitive impairment/hippocampus/xygen-glucose deprivation/reoxygenation/miR-335/ROCK2/NLRP3 inflammasome/stress granule/pyroptosis分类
医药卫生引用本文复制引用
傅晨,巩俐,王翎沣,马承平,刘雪梅..基于miR-335/ROCK2轴探讨补肾启智方对OGD/R损伤HT22细胞焦亡与应激颗粒的影响[J].中医学报,2024,39(8):1583-1590,8.基金项目
国家自然科学基金青年科学基金项目(82104808) （82104808）
