首页|苦杏仁苷对流感病毒FM1诱导的微血管内皮细胞通透性的影响

苦杏仁苷对流感病毒FM1诱导的微血管内皮细胞通透性的影响

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目的 研究苦杏仁苷对甲型流感病毒FM1诱导的微血管内皮细胞(PMVEC)通透性增加的影响及其作用机制.方法 将PMVEC随机分为对照组、模型组(100 TCID50 FM1)、苦杏仁苷低剂量组(100 TCID50 FM1+4.0 mg·mL-1苦杏仁苷)、苦杏仁苷中剂量组(100 TCID50 FM1+8.0 mg·mL-1苦杏仁苷)、苦杏仁苷高剂量组(100 TCID50 FM1+16.0 mg·mL-1苦杏仁苷)及740Y-P组(100 TCID50 FM1+16.0 mg·mL-1 苦杏仁苷+50 μmol·L-1 的 PI3K 激活药740Y-P).以噻唑蓝(MTT)法、Transwell法、酶联免疫吸附测定(ELISA)法、蛋白质印迹法分别检测各组细胞增殖、细胞通透性、炎性因子表达水平和蛋白表达水平.结果 对照组、模型组和苦杏仁苷低、中、高剂量组的白细胞介素-6(IL-6)水平分别为(50.12±3.16)、(93.12±5.61)、(80.33±6.24)、(70.05±5.46)和(61.03±4.68)pg·mL-1,肿瘤坏死因子-α(TNF-α)水平分别为(101.31±9.24)、(167.05±10.31)、(142.02±10.13)、(125.34±9.87)和(112.44±8.05)pg·mL-1.对照组、模型组、苦杏仁苷低剂量组、苦杏仁苷中剂量组、苦杏仁苷高剂量组、740Y-P组细胞跨上皮电阻(TER)分别为(53.01±4.17)、(24.98±2.66)、(30.01±3.49)、(36.84±3.25)、(46.23±4.31)和(30.21±3.16)Ω×cm2,磷酸化的磷脂酰肌醇-3-羟激酶(p-PI3K)蛋白水平分别为 0.34±0.04、1.01±0.09、0.80±0.08、0.61±0.07、0.43±0.05 和0.87±0.09,磷酸化雷帕霉素靶蛋白(p-mTOR)水平分别为0.27±0.03、0.82±0.10、0.60±0.06、0.42±0.03、0.31±0.02和 0.84±0.02.以上指标,模型组和对照组比较,苦杏仁苷低、中、高剂量组分别与模型组比较,苦杏仁苷低、中、高剂量组之间比较,苦杏仁苷高剂量组与740Y-P组比较,在统计学上差异均有统计学意义(均P<0.05).结论 苦杏仁苷可能通过抑制PI3K/AKT/mTOR通路降低流感病毒FM1诱导的PMVEC细胞通透性增加.
Effects of amygdalin on permeability of microvascular endothelial cells induced by influenza virus FM1
Objective To investigate the effect of amygdalin on the permeability increase of microvascular endothelial cells(PMVEC)induced by influenza A virus FM1 and its mechanism.Methods The PMVEC cells were randomly divided into control group,model group(100 TCID50 FM1),amygdalin low-dose group(100 TCID50 FM1+4.0 mg·mL-1 amygdalin),amygdalin medium-dose group(100 TCID50 FM1+8.0 mg·mL-1 amygdalin),amygdalin high-dose group(100 TCID50 FM1+16.0 mg·mL-1 amygdalin)and 740Y-P group(100 TCID50 FM1+16.0 mg·mL-1 amygdalin+50 μmol·L-1 PI3K activator 740Y-P).Methyl thiazolyl tetrazolium(MTT)method,Transwell method,enzyme linked immunosorbent assay(ELISA)method and Western blot method were used to detect cell proliferation,cell permeability,inflammatory factor expression level and protein expression level in each group,respectively.Results Interleukin-6(IL-6)levels in control,model,amygdalin-L,amygdalin-M,amygdalin-H groups were(50.12±3.16),(93.12±5.61),(80.33±6.24),(70.05±5.46)and(61.03±4.68)pg·mL-1,respectively;the levels of tumor necrosis factor-α(TNF-α)in each group were(101.31±9.24),(167.05±10.31),(142.02±10.13),(125.34±9.87)and(112.44±8.05)pg·mL-1,respectively.The cell transepithelial resistance(TER)of control,model,amygdalin-L,amygdalin-M,amygdalin-H and 740Y-P groups were(53.01±4.17),(24.98±2.66),(30.01±3.49),(36.84±3.25),(46.23±4.31),(30.21±3.16)Ω × cm2;phosphorylated phosphatidylinositol-3 hydroxy kinase(p-PI3K)protein levels in each group were 0.34±0.04,1.01±0.09,0.80±0.08,0.61±0.07,0.43±0.05,0.87±0.09,respectively;phosphorylated mammalian target of repamycin(p-mTOR)levels in each group were 0.27±0.03,0.82±0.10,0.60±0.06,0.42±0.03,0.31±0.02 and 1.01±0.12,respectively.Compared model group with control group;compared amygdalinp-L,-M,-H groups with model group;compared amygdalinp-H group with 740Y-P group,the differences of the above indicators were all statistically significant(all P<0.05).Conclusion Amygdalin may decrease the permeability of PMVEC cells induced by influenza virus FM1 by inhibiting PI3K/AKT/mTOR pathway.

amygdalininfluenza A virusmicrovascular endothelial cellpermeabilityphosphatidylinositol-3-hydroxykinase/protein kinase B/mammalian target of rapamycin pathway

王祎、曲书焱、姜琪琦、钟燕春

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河北中医药大学基础医学院温病学教研室,河北石家庄 050200

苦杏仁苷 甲型流感病毒 微血管内皮细胞 通透性 磷脂酰肌醇-3-羟激酶/蛋白质激酶B/雷帕霉素靶蛋白通路

国家自然科学基金

81774195

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(6)
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