miR-361介导PI3K/Akt/Ca^(2+)通路对大鼠肥大细胞增殖情况及组胺与IL-4表达水平的影响  

The effects of miR-361 mediated PI3K/Akt/Ca^(2+)pathway on proliferation of rat mast cells and expression levels of histamine and IL-4

作　　者：蒙金秋 简华慧[1] 蒙安定[1] 陈丽[2] 林淼鑫[2] 周伟杰[2] 黄爱良 罗慧秋 林峻慰 唐秀生[3] Meng Jinqiu;Jian Huahui;Meng Anding;Chen Li;Lin Miaoxin;Zhou Weijie;Huang Ailiang;Luo Huiqiu;Lin Junwei;Tang Xiusheng(Dermatology Department,People's Hospital of Baise City,Baise,Guangxi 533000,China;Clinical Laboratory,People's Hospital of Baise City,Baise,Guangxi 533000,China;Dermatology Department,Youjiang Medical College for Nationalities,Baise,Guangxi 533000,China)

机构地区：[1]百色市人民医院皮肤科,广西百色533000 [2]百色市人民医院检验科,广西百色533000 [3]右江民族医学院附属医院皮肤科,广西百色533000

出　　处：《医药前沿》2024年第10期1-3,7,共4页Journal of Frontiers of Medicine

基　　金：百色市科学研究与技术开发计划项目(百科20193212)。

摘　　要：目的:探讨miR-361介导PI3K/AKt/Ca^(2+)通路对活化大鼠肥大细胞增殖情况及组胺与白细胞介素-4(IL-4)表达水平的影响。方法:将大鼠肥大细胞(RBL-2H3细胞)分为miR-361过表达组、miR-361抑制组、miR-361对照组和正常对照组,miR-361过表达组、miR-361抑制组、miR-361对照组细胞分别转染miR-361过表达、miR-361抑制及miR-361无义序列寡核苷酸片段,测定并比较各组细胞miR-361 mRNA、组胺、IL-4及PI3K/AKt/Ca^(2+)通路相关蛋白水平。结果:miR-361过表达组细胞miR-361 mRNA相对表达水平高于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。CCK-8法检测结果显示,miR-361过表达组24、48、72 h细胞增值率均低于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。ELISA法检测结果显示,miR-361过表达组细胞组胺和IL-4水平的表达水平低于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。Western印迹法检测结果显示,miR-361过表达组细胞p-AKT、AKT、p-PI3K、PI3K蛋白表达水平均高于miR-361抑制组、miR-361对照组及正常对照组,差异均有统计学意义(P<0.05)。结论:miR-361过表达可抑制大鼠肥大细胞的增殖,降低组胺与IL-4的表达水平,其作用机制可能与其能够提高PI3K/AKt/Ca^(2+)通路活性有关。Objective To investigate the effect of miR-361 mediating the PI3K/AKt/Ca^(2+)pathway on the proliferation of activated rat mast cells and the expression levels of histamine and interleukin-4(IL-4).Methods Rat mast cells(RBL-2 H3 cells)were divided into miR-361 overexpression group,miR-361 inhibition group,miR-361 control group and normal control group.miR-361 overexpression group,miR-361 inhibition group and miR-361 control group cells were transfected with miR-361 overexpression,miR-361 inhibition and miR-361 nonsense oligonucleotide fragments respectively.Sequence oligonucleotide fragments,miR-361 mRNA,histamine,IL-4 and PI3K/AKt/Ca^(2+)pathway-related protein levels were determined and compared in each group.Results The relative expression level of miR-361 mRNA in the miR-361 overexpression group was higher than that in the miR-361 inhibition group,miR-361 control group,and normal control group,and the differences were statistically significant(P<0.05).The CCK-8 assay results showed that the cell proliferation rates at 24,48,and 72 h in the miR-361 overexpression group were lower than those in the miR-361 inhibition group,miR-361 control group,and normal control group,and the differences were statistically significant(P<0.05).The ELISA assay results showed that the expression levels of histamine and IL-4 in the miR-361 overexpression group were lower than those in the miR-361 inhibition group,miR-361 control group,and normal control group,and the differences were statistically significant(P<0.05).The Western blot results showed that the protein expression levels of p-AKT,AKT,p-PI3K,and PI3K in the miR-361 overexpression group were higher than those in the miR-361 inhibition group,miR-361 control group,and normal control group,and the differences were statistically significant(P<0.05).Conclusions Overexpression of miR-361 inhibited the proliferation of rat mast cells and reduced the expression levels of histamine and IL-4,and its mechanism of action may be related to its ability to increase the activity of the

关 键 词：miR-361 肥大细胞 组胺 白细胞介素-4 PI3K/AKt/Ca^(2+)通路 

分 类 号：R758.24[医药卫生—皮肤病学与性病学]