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目的探讨大麻素Ⅱ型受体(CB2)激动剂AM1241对大鼠肝星状细胞系(HSC-T6)影响及其机制。方法将HSC-T6细胞随机分为对照组、氧化应激组及20、80μmol/L AM1241干预组;对照组正常培养,氧化应激组用含100 m U/L葡萄糖氧化酶(GO)完全培养液培养2 h;AM1241干预组分别加入20、80μmol/L的AM1241干预3 h后,再加入100 m U/L GO干预2 h;细胞免疫化学染色法检测HSC-T6细胞中α-平滑肌肌动蛋白(α-SMA)表达;酶联免疫法检测HSC-T6细胞培养液上清中I型胶原(Col I);硫代巴比妥酸法检测丙二醛含量;氮蓝四唑法检测超氧化物歧化酶(SOD)活力;Western blot检测HSC-T6细胞核转录相关因子(Nrf2)总蛋白与核蛋白含量。结果与对照组比较,氧化应激组HSC-T6细胞胞浆中α-SMA、Col I表达量均增多(P<0.05);与氧化应激组比较,AM1241干预组HSC-T6细胞胞浆中α-SM A、Col I表达量均减少(P<0.05);与对照组比较,氧化应激组HSC-T6细胞中丙二醛含量上升,SOD活性下降(P<0.05);与氧化应激组比较,AM1241干预组HSC-T6细胞丙二醛含量下降,SOD活性上升(P<0.05);对照组、氧化应激组及20、80μmol/L AM1241干预组HSC-T6细胞中Nrf2总蛋白表达量分别为(0.62±0.021)、(0.60±0.015)、(0.59±0.020)、(0.61±0.032),各组差异无统计学意义(P>0.05);与氧化应激组比较,AM1241干预组Nrf2核蛋白表达量均增加(P<0.05)。结论大麻素CB2受体激动剂AM1241可抑制氧化应激作用下HSC-T6细胞活化,其机制可能与AM1241促进HSC-T6细胞中Nrf2发生核内转移,增加抗氧化作用有关。
Objective To investigate the effect and mechanism of AM1241, a cannabinoid type Ⅱ receptor (CB2) agonist, on rat hepatic stellate cell line (HSC-T6). Methods HSC-T6 cells were randomly divided into control group, oxidative stress group and 20, 80μmol / L AM1241 intervention group. The control group was cultured normally and the oxidative stress group was completely cultured with 100 mU / L glucose oxidase (GO) Liquid culture for 2 h. The AM1241 intervention group was treated with 20, 80 μmol / L AM1241 for 3 h, followed by 100 m U / L GO for 2 h. The α-SMA activity in HSC-T6 cells was detected by immunocytochemical staining The expression of α-SMA was detected by enzyme-linked immunosorbent assay (ELISA), the type I collagen (Col I) in culture supernatant of HSC-T6 cells was detected by ELISA, the content of malondialdehyde was detected by thiobarbituric acid method, Oxidative dismutase (SOD) activity; Western blot was used to detect the total protein and nucleoprotein of nuclear transcription related factor (Nrf2) in HSC-T6 cells. Results Compared with the control group, the expressions of α-SMA and Col I in HSC-T6 cells were increased in the oxidative stress group (P <0.05). Compared with the oxidative stress group, the expressions of α-SMA and Col I in the cytoplasm of HSC- (P <0.05). Compared with the control group, the content of malondialdehyde (MDA) in HSC-T6 cells increased and the activity of SOD decreased (P <0.05) in the oxidative stress group. Compared with the oxidative stress Compared with the control group, the expression of Nrf2 protein in HSC-T6 cells was significantly decreased in AM1241-treated HSC-T6 cells as compared with the control group, oxidative stress group and 20, 80μmol / L AM1241 intervention group (0.62 ± 0.021), (0.60 ± 0.015), (0.59 ± 0.020) and (0.61 ± 0.032), respectively. There was no significant difference between groups (P> 0.05). Compared with oxidative stress group, AM1241 intervention group Nrf2 nuclear protein expression increased (P <0.05). CONCLUSION: The cannabinoid CB2 receptor agonist AM1241 can inhibit the activation of HSC-T6 cells under oxidative stress. The possible mechanism is that AM1241 may promote the intranuclear transfer of Nrf2 in HSC-T6 cells and increase the anti-oxidative effect.