连翘苷调控miR-146a对脂多糖诱导的肾小管上皮细胞HK-2凋亡及氧化应激的影响  

Effect of forsythin on lipopolysaccharide-induced apoptosis and oxidative stress in renal tubular epithelial cells HK-2 by regulating miR-146a

作　　者：梁勇[1] 武军元[2] 刘禹赓[1] 刘波[1] 魏兵[1] 张向群[1] 曾红[1] LIANG Yong;WU Jun-Yuan;LIU Yu-Gen(Department of Emergency,West District,Chaoyang Hospital,Capital Medical University,Beijing 100043,China)

机构地区：[1]首都医科大学附属北京朝阳医院西区急诊科,北京100043 [2]首都医科大学附属北京朝阳医院急诊科,北京100043

出　　处：《中国老年学杂志》2022年第10期2438-2443,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81801882)。

摘　　要：目的探讨连翘苷对脂多糖(LPS)诱导的肾小管上皮细胞(HK)-2凋亡和氧化应激的影响。方法采用1μg/ml LPS干预对数期HK-2细胞6 h记为模型(Model)组。Con组正常培养细胞,不用LPS干预。采用不同浓度(5、10、20 ng/ml)的连翘苷处理LPS诱导的HK-2细胞,依次记为实验1、2、3组。细胞计数试剂盒(CCK)-8法、流式细胞术分别检测细胞活力和凋亡。检测超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH)活性及丙二醛(MDA)含量。实时荧光定量-聚合酶链反应(RT-qPCR)检测miR-146a表达水平。采用上述方法检测过表达miR-146a对LPS诱导的HK-2细胞凋亡和氧化应激的影响。结果与Con组比较,Model组HK-2细胞活力、miR-146a表达、SOD和GSH活性显著降低,凋亡率、MDA含量显著升高(P<0.05)。与Model组比较,实验2组、实验3组HK-2细胞活力、miR-146a表达、SOD和GSH活性显著升高,凋亡率、MDA含量显著降低(P<0.05)。过表达miR-146a可提高LPS作用下细胞活力,减轻LPS诱导HK-2细胞凋亡的和氧化应激损伤(P<0.05)。抑制miR-146a表达可降低连翘苷对LPS诱导HK-2细胞凋亡的和氧化应激损伤的影响(P<0.05)。结论连翘苷通过上调miR-146a表达进而减轻对LPS诱导的细胞凋亡和氧化应激损伤。Objective To investigate the effect of forsythin on lipopolysaccharide(LPS)-induced apoptosis and oxidative stress in renal tubular epithelial cells(HK-2)and its possible mechanism.Methods The logarithmic HK-2 cells were treated with 1μg/ml LPS for 6 h and recorded as model group.Normal cells were cultured without LPS intervention in control group.Different concentrations(5,10,20 ng/ml)of forsythin were used to treat HK-2 cells induced by LPS.Cell counting kit(CCK-8)method and flow cytometry were used to detect cell viability and apoptosis.The kit was applied to detect the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH)and the content of malondialdehyde(MDA).The expression level of miR-146a was measured by real-time quantitative PCR(RT-qPCR).The effect of miR-146a overexpression on LPS-induced HK-2 cell apoptosis and oxidative stress were detected by the above method.Results Compared with the control group,the cell viability,miR-146a expression,SOD and GSH activities of HK-2 cells in the model group were significantly reduced,whereas the apoptosis rate and MDA content were significantly increased(P<0.05).Compared with the model group,the cell viability,miR-146a expression,SOD and GSH activities of HK-2 cells in the 10 ng/ml and 20 ng/ml forsythin intervention groups were significantly increased,whereas the apoptosis rate and MDA content were significantly reduced(P<0.05).miR-146a overexpression could increase cell viability under the action of LPS,and reduce LPS-induced HK-2 cell apoptosis and oxidative stress damage(P<0.05).The expression of miR-146a inhibition could reduce the effect of forsythin on LPS-induced HK-2 cell apoptosis and oxidative stress damage(P<0.05).Conclusions Forsythoside could reduce LPS-induced apoptosis and oxidative stress damage by up-regulating miR-146a expression.

关 键 词：连翘苷 肾小管上皮细胞 MIR-146A 凋亡 氧化应激损伤 

分 类 号：R692[医药卫生—泌尿科学]