摘要
目的探讨木通皂苷D对脂多糖(LPS)诱导的人脑微血管内皮细胞(HBMEC)凋亡和氧化应激的影响及作用机制。方法体外培养HBMEC,用不同剂量(12.5、25、50μmol/L)木通皂苷D干预LPS诱导的HBMEC 24 h后,CCK-8法检测细胞增殖活性,流式细胞术检测细胞凋亡,比色法检测细胞中SOD活性和MDA含量,qRT-PCR检测细胞中circ_TTC3表达。转染circ_TTC3小干扰RNA或过表达载体至HBMEC细胞,上述相同方法观察circ_TTC3对LPS诱导的HBMEC凋亡和氧化应激的影响。结果与LPS组比较,木通皂苷D降低了LPS诱导的HBMEC增殖抑制率、凋亡率及细胞中MDA含量(P<0.05),而提高了SOD活性(P<0.05)。敲减circ_TTC3降低了LPS诱导的HBMEC增殖抑制率、凋亡率及细胞中MDA含量(P<0.05),而提高了SOD活性(P<0.05)。木通皂苷D降低了LPS诱导的HBMEC中circ_TTC3表达,过表达circ_TTC3逆转了木通皂苷D对LPS诱导的HBMEC增殖、凋亡及氧化应激的影响。结论木通皂苷D可能通过下调circ_TTC3抑制LPS诱导的HBMEC凋亡和氧化应激。
Objective To investigate the effect of Akebia saponin D on apoptosis and oxidative stress of human brain microvascular endothelial cells(HBMEC)induced by lipopolysaccharide(LPS)and its possible mechanism.Methods HBMEC is cultured in vitro and intervened with different doses(12.5,25,50μmol/L)of Akebia Saponin D and LPS for 24 h,and then cell proliferation activity is detected by CCK-8 method;cell apoptosis is detected by flow cytometry.The activity of SOD and the content of MDA in cells are detected by colorimetric method.qRT-PCR is used to detect the expression of circ_TTC3.circ_TTC3 siRNA or overexpression vector is transfected into HBMEC cells,and then the same methods described above are used to observe the effect of circ_TTC3 on apoptosis and oxidative stress of HBMEC induced by LPS.Results Compared with the LPS group,Akebia saponin D reduced the LPS-induced HBMEC proliferation inhibition rate,apoptosis rate and the content of MDA in cells(P<0.05),but increased the activity of SOD(P<0.05).Knock down of circ_TTC3 reduced the LPS-induced HBMEC proliferation inhibition rate,apoptosis rate and the content of MDA in cells(P<0.05),but increased the activity of SOD(P<0.05).Akebia saponin D reduced the expression of circ_TTC3 in HBMEC induced by LPS,and overexpression of circ_TTC3 reversed the effect of Akebia saponin D on the proliferation,apoptosis and oxidative stress of HBMEC induced by LPS.Conclusion Akebia saponin D may inhibit LPS-induced HBMEC apoptosis and oxidative stress by down-regulating circ_TTC3.
作者
郗欧
刘禹岐
马进
焦富英
阮林
XI Ou;LIU Yu-qi;MA Jin;JIAO Fu-ying;RUAN Lin(The First Department of Encephalopathy,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China;The Second Department of Cardiology,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China)
出处
《现代中药研究与实践》
CAS
2022年第2期28-32,共5页
Research and Practice on Chinese Medicines
基金
辽宁省教育厅重点攻关项目(L202054)。
