| 陈宝磊,高映春,吴磊.枇杷叶提取物通过激活磷脂酰肌醇 3激酶 /蛋白激酶 B信号通路减轻脂多糖诱导的肺细胞损伤[J].安徽医药,2022,26(3):453-457. |
| 枇杷叶提取物通过激活磷脂酰肌醇 3激酶 /蛋白激酶 B信号通路减轻脂多糖诱导的肺细胞损伤 |
| Extract of loquat [Eriobotrya japonica (Thunb) Lindl] leaves reduces LPS-induced cell damage in a lung injury model by activating PI3K/Akt signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2022.03.007 |
| 中文关键词: 急性肺损伤 枇杷叶提取物 磷脂酰肌醇 3激酶 /蛋白激酶 B信号通路 脂多糖 A549细胞 增殖 凋亡 |
| 英文关键词: Acute lung injury Extract of loquat [Eriobotrya japonica (Thunb) Lindl] leaves PI3K/Akt signaling pathway Lipopolysaccharide A549 cells Proliferation Apoptosis |
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| 中文摘要: |
| 目的探讨枇杷叶提取物对脂多糖( LPS)诱导的人 Ⅱ型肺泡上皮细胞 A549增殖、凋亡及氧化应激的影响及其可能作用机制。方法 2019年 6月至 2020年 7月，采用 LPS诱导的 A549细胞建立肺损伤模型( LPS组)同时将正常培养的细胞作为对照组。不同剂量( 1 mg/L、2 mg/L、4 mg/L)的枇杷叶提取物处理细胞( LPS+枇杷叶 -L组、 LPS+枇，杷叶 -M组、 LPS+枇杷叶 -H组)，添加磷脂酰肌醇 3激酶( PI3K)/蛋白激酶 B(Akt)信号通路抑制剂 LY294002与枇杷叶提取物处理细胞( LPS+枇杷叶 -H+ LY294002组)；采用 MTT法检测细胞增殖；采用流式细胞术检测细胞凋亡率；采用 2，4-二硝基苯肼显色法检测乳酸脱氢酶(LDH)的含量；采用黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)的含量；采用硫代巴比妥酸法检测丙二醛的含量；蛋白质印迹法( Western blotting)检测磷酸化磷脂酰肌醇 -3-激酶( p-PI3K)、磷酸化蛋白激酶 B(p-Akt)、 B细胞淋巴瘤 -2(Bcl-2)、 Bcl-2相关 X(Bax)蛋白蛋白表达量。结果与对照组比较， LPS组吸光度［( 1.38±0.06)比( 0.55±0.02)］及 SOD的含量［( 81.66±5.36)U/L比(14.65±1.06)U/L］降低，凋亡率［(6.41±0.25)%比( 27.43±1.00)%］、 Bax蛋白水平及 LDH［(242.86±6.09)U/L比( 875.92±15.01)U/ L］、丙二醛［( 121.55±3.17)μmol/g比( 424.46±6.48)μmol/g］的含量升高， Bcl-2、p-PI3K［( 0.60±0.04)比( 0.13±0.01)］、 p-Akt［( 0.51±0.04)比( 0.09±0.01)］蛋白水平降低，差异有统计学意义( P<0.05)；与 LPS组比较， LPS+枇杷叶 -M组、 LPS+枇杷叶 -H组吸光度［( 0.55±0.02)比( 0.86±0.03)、(1.18±0.05)］及 SOD［( 14.65±1.06)U/L比( 36.84±2.08)U/L、(70.97±3.03)U/L］的含量升高，凋亡率［( 27.43±1.00)%比( 22.24±0.81)%、(14.78±0.49)%］、 Bax蛋白水平及 LDH［( 875.92±15.01)U/L比( 641.95±9.81)U/L、(365.47±7.02)U/L］、丙二醛［( 424.46±6.48)μmol/g比( 277.94±6.90)μmol/g、(156.30±3.26)μmol/g］的含量降低， Bcl-2、p-PI3K［( 0.13±0.01)比( 0.32±0.01)、(0.54±0.03)］、 p-Akt［( 0.09±0.01)比( 0.25±0.02)、(0.42±0.03)］蛋白水平升高，均差异有统计学意义( P<0.05)；添加 LY294002可明显逆转枇杷叶提取物对 LPS诱导的 A549细胞增殖、凋亡及氧化应激的作用。结论枇杷叶提取物可通过激活 PI3K/Akt信号通路减轻 LPS诱导的 A549细胞增殖抑制、凋亡和氧化应激效应，为探究枇杷叶治疗细菌感染 |
| 英文摘要: |
| Objective To explore the effect of extract of loquat [Eriobotrya japonica (Thunb) Lindl] leaves on the proliferation, apoptosis and oxidative stress of human type Ⅱ alveolar epithelial cells A549 induced by lipopolysaccharide (LPS) and its possible mechanism.Methods LPS-induced A549 cells were used to establish a lung injury model (LPS group) from June 2019 to July 2020, and conventionally cultured cells were used as controls. Different doses of extract of loquat [Eriobotrya japonica (Thunb) Lindl] leaves (1 mg/L, 2 mg/L, 4 mg/L) were used to treat cells (LPS+loquat [Eriobotrya japonica (Thunb) Lindl] leaves-L group, LPS+loquat [Eriobotrya japon? ica (Thunb) Lindl] leaves-M group, and LPS+loquat [Eriobotrya japonica (Thunb) Lindl] leaves-H group). Phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway inhibitor LY294002 was added with the extract of loquat [Eriobotrya japonica (Thunb) Lindl] leaves to treat cells (LPS+loquat [Eriobotrya japonica (Thunb) Lindl] leaves-H+LY294002 group). The MTT method was used todetect cell proliferation. Flow cytometry was used to detect the apoptosis rate. The 2,4-dinitrophenylhydrazine color method was used todetect the content of lactate dehydrogenase (LDH). Xanthine oxidase method was used to detect the content of superoxide dismutase(SOD). The thiobarbituric acid method was used to detect the content of malondialdehyde (MDA). Western blotting method was used todetect the protein expressions in phosphorylated phosphatidylinositol-3-kinase (p-PI3K), phosphorylated protein kinase B (p-Akt), B-cell lymphoma-2 (Bcl-2), and Bcl-2-related X protein (Bax).Results Compared with the control group, the optical density (OD) value [(1.38±0.06) vs. (0.55±0.02)] and the content of SOD [(81.66±5.36) U/L vs. (14.65±1.06) U/L] were decreased, the apoptosis rate [(6.41± |
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