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

作　　者：刘云婷 傅晨[1] 刘嘉 巩俐 杨小蕊 刘雪梅[1] LIU Yunting;FU Chen;LIU Jia;GONG Li;YANG Xiaorui;LIU Xuemei(Dongfang Hospital Affiliated to Beijing University of Chinese Medicine,Beijing China 100078)

机构地区：[1]北京中医药大学东方医院,北京100078

出　　处：《中医学报》2024年第8期1601-1608,共8页Acta Chinese Medicine

基　　金：国家自然科学基金项目(81973788)。

摘　　要：目的:观察清脑滴丸对大鼠脑微血管内皮细胞(rat brain microvascular endothelial cells,RBMECs)氧糖剥夺/复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤模型的影响,探讨其调节长链非编码RNA母系表达基因3(long non-coding RNA maternally expressed gene 3,LncRNA MEG3)表达及对核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体活化的作用。方法:将20只SD大鼠随机分为空白血清组和清脑滴丸含药血清组(150 mg·kg^(-1)),每组10只,连续灌胃7 d,每日2次,从而制备血清样品。取对数生长期的RBMECs,随机分为对照组、模型组、清脑滴丸组、MEG3过表达组、MEG3过表达+清脑滴丸组。除对照组外,其余各组细胞均进行OGD/R造模。观察细胞血管形成情况,采用CCK-8法检测细胞存活率,qRT-PCR法检测细胞LncRNA MEG3 mRNA表达水平,Western blot法检测细胞NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a caspase recruitment domain,ASC)、活化的半胱氨酸天冬氨酸蛋白酶1(cleaved cysteinyl aspartate specific proteinase-1,c-Caspase-1)、肿瘤抑制蛋白p53(tumor suppressor protein p53,p53)、活化的半胱氨酸天冬氨酸蛋白酶3(cleaved cysteinyl aspartate specific proteinase-3,c-Caspase-3)表达水平。结果:与对照组比较,模型组细胞存活率降低、RBMECs形成的血管总长度缩短(P<0.01);与模型组比较,清脑滴丸组、MEG3过表达+清脑滴丸组细胞存活率升高、RBMECs形成的血管总长度增加(P<0.05)。与对照组比较,模型组LncRNA MEG3 mRNA水平升高(P<0.05);与模型组比较,清脑滴丸组LncRNA MEG3 mRNA水平降低(P<0.01),MEG3过表达+清脑滴丸组、MEG3过表达组LncRNA MEG3 mRNA水平升高(P<0.01);与清脑滴丸组比较,MEG3过表达+清脑滴丸组细胞LncRNA MEG3 mRNA水平升高(P<0.01)。与对照组比较,模型组细胞NLRP3、ASC、c-Caspase-1、p53、c-Caspase-3蛋白表达水平升高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 Qingnao 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 Dr

关 键 词：清脑滴丸 大鼠脑微血管内皮细胞 OGD/R LncRNA MEG3 NLRP3 

分 类 号：R285.5[医药卫生—中药学]