大麻素CB2受体激动剂AM1241对肝星状细胞影响  

Influence of cannabinoid CB2 receptor agonist AM1241 on cultured hepatic stellate cells

作　　者：史云[1] 何萍[1] 孙建超[1] 王豫萍[1] 

机构地区：[1]贵州医科大学医学检验学院,贵州贵阳550004

出　　处：《中国公共卫生》2016年第5期613-617,共5页Chinese Journal of Public Health

基　　金：国家自然科学基金(81460125);贵州省留学人员科技活动项目经费资助(201505);贵州省科技厅中药现代化项目(黔科合ZY字[2012]3017);贵州省省长基金(黔省专合字2012[44])

摘　　要：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 study the influence of cannabinoid CB2 receptor agonist AM1241 on cultured hepatic stellate cell line（HSC-T6）. Methods HSC-T6 cells were divided into a control group（normal culture） ,an oxidative stress model group（ treated with 100 mU/L glucose oxidase）, and two AM1241 treatment groups（ first treated with 20 or 80 μmol/L AM1241 and then treated with 100 mU/L glucose oxidase）. Alpha-smooth muscle actin（a-SMA） expression in HSC-T6 cells was detected with imrnunocytochemical method. Collagen type I（ Col I） content in supernatants of HSC- T6 cell culture,malondialdehyde （MDA） and superoxide dismutase （SOD） contents in HSC-T6 cells were determined with enzyme-linked immunosorbent assay （ELISA） ;thiobarbituric acid method for MDA and nitroblue tetrazolium meth- od for SOD were adopted in the detections. Western blot was used to detect total protein and nuclear protein of nuclear factor erythroid-2 related factor 2 （Nrf2） in HSC-T6 cells. Results Compared with the control group, the expression of Col 1 and a-SMA in HSC-T6 cells were significantly increased in the oxidative stress group（ P 〈 0.05 ） and the expression of Col I and a-SMA in HSC-T6 cells were significantly decreased （P 〈 0. 05）. Compared with the control group, MDA of the oxidative stress group increased but SOD decreased significantly（ both P 〈 0. 05 ） ; compared with the oxidative stress group, the content of SOD in AM1241 treatment group decreased but MDA increased significantly （both P 〈 0. 05 ）. The expression of Nrf2 total protein in the control, oxidative stress and two AM1241 treatment groups were 0. 57 ± 0. 021,0. 60 ± 0. 015,0. 59 ±0. 020, and 0. 61 ± 0. 032, respectively, without significant difference among the groups. No expression of nuclear protein of Nrf2 was detected in the control group; there was the expression of Nrf2 nuclear protein in the oxidative stress group and the expression of Nrf2 nuclear protein in the AM1241treatement increased significantly （

关 键 词：大麻素Ⅱ型受体(CB2) 激动剂AM1241 大鼠肝星状细胞系(HSC-T6) 氧化应激 核转录相关因 子(Nrf2) 

分 类 号：R512.6[医药卫生—内科学]