| 宋云飞.异丙酚抑制内质网应激减轻脂多糖诱导人肝细胞损伤的机制研究[J].安徽医药,2021,25(3):451-455. |
| 异丙酚抑制内质网应激减轻脂多糖诱导人肝细胞损伤的机制研究 |
| Study on the mechanism of propofol on lipopolysaccharide injured human hepatocytes by inhibiting endoplasmic reticulum stress |
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| DOI：10.3969/j.issn.1009-6469.2021.03.006 |
| 中文关键词: 异丙酚 脂多糖 化学性与药物性肝损伤 内质网应激 丙氨酸转氨酶 白细胞介素 -6 NF-κB 激活转录因子 6 |
| 英文关键词: Propofol Lipopolysaccharide Chemical and drug induced liver injury ER stress Alanine transaminase Interleukin-6 NF-kappa B Activating transcription factor 6 |
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| 中文摘要: |
| 目的探讨异丙酚(PPF)对脂多糖(LPS)诱导人正常肝细胞 L-02损伤的保护作用。方法用不同浓度的 LPS来制备人肝细胞损伤模型，将人正常肝细胞 L-02按随机数字表法分为五组:正常组、 LPS组、 LPS+25、50 μmol/L PPF组以及 LPS+4 mmol/ L4-苯基丁酸( PBA)组。四甲基偶氮唑盐微量酶反应比色法( MTT法)检测 L-02细胞的存活率，酶联免疫吸附测定( ELISA)测定各组细胞上清中丙氨酸氨基转移酶( ALT)和天冬氨酸氨基转移酶( AST)的活性。实时荧光定量逆转录聚合酶链反应( qRTPCR)检测各组细胞中炎性因子白细胞介素( IL)-1β、IL-6和肿瘤坏死因子 α(TNF-α)mRNA的表达；蛋白质印迹法( Western blotting)检测各组细胞中核因子 -κB(NF-κB)p65、NF-κB、p-肌醇依赖酶 1α(IRE1α)、 IRE1α和激活转录因子 6(ATF-6)蛋白的表达。结果用不同浓度的 LPS处理的人肝细胞存活能力显著降低，且 LPS致人肝细胞损伤的最佳造模条件为 20 mg/L孵育 24 h。与正常组比较， LPS组的人肝细胞存活能力显著降低，细胞数量减少且核固缩；细胞上清中 ALT和 AST的释放量分别增加［( 69.17±5.51)mg/L和( 50.23±5.81)mg/L比( 19.78±1.79)mg/L和( 9.79±0.96)mg/L，P<0.05］； IL-1β、IL-6和 TNF-α mRNA表达显著升高； NF-κB p65表达量明显增强，且内质网应激相关蛋白 p-IRE1α/IRE1α和 ATF-6的表达显著增高［( 2.24±0.28)和(2.55±0.16)比( 1.00±0.17)和( 1.00±0.26)P<0.05］。与 LPS组相比，给予低高剂量 PPF能显著提高人肝细胞的存活率，降低 ALT［(55.30±6.92)mg/L和( 33.70±4.08)mgL］和 AST［(39.46±2.56)mg/L和( 27.62±2.12)mg/L］的释放量；显著降低 IL-1β、IL-6和 TNF-α mRNA的表达；抑制 NF-κB p65及内质网应激相关蛋白 ATF-6和 p-IRE1α/IRE1α蛋白水平的表达。结论 PPF对 LPS诱导的人肝细胞损伤具有一定的保护作用，可能是通过抑制内质网应激从而发挥抗炎作用来实现的。 |
| 英文摘要: |
| Objective To investigate the protective effect of propofol (PPF) on injured human hepatocytes induced by lipopolysac? charide (LPS).Methods The injured hepatocytes models were established according to different concentrations of LPS. Human nor? mal hepatocytes L-02 were randomly divided into 5 groups according to the random number table method: normal group, LPS group,LPS+25, 50 μmol/L PPF group and LPS+4 mmol/L 4-phenylbutyric acid (PBA) group. MTT was used to detect the survival rate of hu?man hepatocytes, and the activities of Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) were determined by en?zyme linked immunosorbent assay (ELISA). The mRNA expression of Interleukin-1(IL)-1β, IL-6 and tumor necrosis factor (TNF)-α was detected by reverse transcription-Polymerase Chain Reaction (qRT-PCR), and the protein expression of nuclear factor kappa-B (NF-κB) p65、 NF-κB、 p-inositol-requiring enzyme (IRE1)α、 IRE1α and activating transcription factor (ATF)-6 was detected by Western blotting.Results The viability of injured human hepatocytes induced by different concentrations of LPS was significantly reduced, andthe most optimum condition was 20 mg/L, culturing for 24 h. Compared with the normal group, the survival ability of human hepato?cytes in LPS group was significantly reduced, the number of cells was decreased and nuclear pyknosis was observed; the release of ALTand AST was increased [(69.17±5.51) mg/L and (50.23±5.81) mg/L vs. (19.78±1.79) mg/L and (9.79±0.96) mg/L,P<0.05]; the expres? sion of IL-1β, IL-6 and TNF-α mRNA was significantly increased; the expression of NF-κB p65 was significantly increased, and the ex? pression of p-IRE1α/IRE1α and ATF-6 were significantly increased [(2.24±0.28) and (2.55±0.16) vs. (1.00±0.17) and (1.00±0.26),P< 0.05]. Compared with LPS group, low and high dose of PPF can significantly improve the survival rate of human hepatocytes, reduce therelease of ALT [(55.30±6.92) mg/L and (33.70±4.08) mg/L] and AST [(39.46±2.56) mg/L and (27.62±2.12) mg/L]; significantly reducethe expression of IL-1β, IL-6 and TNF-α mRNA; inhibit the expression of NF-κB p65 and endoplasmic reticulum (ER) stress-related protein ATF-6 and p-IRE1α/IRE1α protein.Conclusion PPF has a protective effect on LPS induced injury of human hepatocytes, |
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