中医学报2024，Vol.39Issue(8)：1601-1608,8.DOI:10.16368/j.issn.1674-8999.2024.08.264
清脑滴丸通过调节LncRNA MEG3和NLRP3炎症小体对大鼠脑微血管内皮细胞OGD/R损伤的影响
Effect of Qingnao Dropping Pills on OGD/R Injury in Rat Brain Microvascular Endothelial Cells by Regulating LncRNA MEG3 and NLRP3 Inflammasome
摘要
Abstract
Objective:To observe the effect of Qingnao Dropping Pills on oxygen-glucose deprivation/reoxygenation(OGD/R)injury model of rat brain microvascular endothelial cells(RBMECs),and to explore its regulation of long non-coding RNA maternally expressed gene 3(LncRNA MEG3)expression and its role in the activation of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome.Methods:A total of 20 SD rats were randomly divided into a blank serum group and a Qing-nao Dropping Pills containing serum group(150 mg·kg-1),with 10 rats in each group.The rats were treated by gavage for 7 days,twice a day,to prepare serum samples.RBMECs in logarithmic growth phase were randomly divided into a control group,a model group,a Qingnao Dropping Pills group,a MEG3 overexpression group,and a MEG3 overexpression+Qingnao Dropping Pills group.Except for the control group,the cells in the other groups were subjected to OGD/R.The cell angiogenesis was observed and cell survival rate was detected by CCK-8 method.The expression level of LncRNA MEG3 mRNA was detected using qRT-PCR.Western blot was used to detect NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),cleaved cysteinyl aspartate specific proteinase-1(c-Caspase-1),tumor suppressor protein p53(p53)and cleaved cysteinyl aspartate specific proteinase-3(c-Caspase-3).Results:Compared with that of the control group,the cell survival rate was decreased,and the total length of blood vessels formed by RBMECs was significantly shortened in the model group(P<0.01).Compared with that of the model group,the cell survival rate and the total length of blood vessels formed by RBMECs were increased significantly in the Qingnao Dropping Pill group and MEG3 overexpression+Qingnao Dropping Pill group(P<0.05).Compared with the control group,the mRNA level of LncRNA MEG3 in the model group was increased significantly(P<0.05).Compared with the model group,the expression level of LncRNA MEG3 mRNA was significantly decreased in the Qingnao Dropping Pills group(P<0.01),and the expression level of LncRNA MEG3 mRNA was significantly increased in the MEG3 overexpression+Qingnao Dropping Pills group and MEG3 overexpression group(P<0.01).Compared with the Qingnao Dropping Pills group,the mRNA level of LncRNA MEG3 was significantly increased in the MEG3 overexpression+Qingnao Dropping Pills group(P<0.01).Compared with the control group,the expression levels of NLRP3,ASC,c-Caspase-1,p53 and c-Caspase-3 in the model group were increased significantly(P<0.01).Compared with the model group,the protein expression levels of NLRP3,ASC,c-Caspase-1,p53 and c-Caspase-3 in the Qingnao Dropping Pills group and MEG3 overexpression+Qingnao Dropping Pills group were decreased significantly(P<0.05).Compared with the Qingnao Dropping Pills group,the protein expression levels of NLRP3,ASC,c-Caspase-1 and p53 were significantly increased in the MEG3 overexpression+Qingnao Dropping Pills group(P<0.05).Conclusions:Qingnao Dropping Pills can protect RBMECs by down-regulating the expres-sion of LncRNA MEG3 and inhibiting the activation of NLRP3 inflammasome to improve cell survival rate and angiogenesis ability.关键词
清脑滴丸/大鼠脑微血管内皮细胞/OGD/R/LncRNA MEG3/NLRP3Key words
Qingnao Dropping Pill/rat brain microvascular endothelial cell/OGD/R/LncRNA MEG3/NLRP3分类
医药卫生引用本文复制引用
刘云婷,傅晨,刘嘉,巩俐,杨小蕊,刘雪梅..清脑滴丸通过调节LncRNA MEG3和NLRP3炎症小体对大鼠脑微血管内皮细胞OGD/R损伤的影响[J].中医学报,2024,39(8):1601-1608,8.基金项目
国家自然科学基金项目(81973788) （81973788）
