天麻素对大鼠骨骼肌细胞化学损伤的保护

背景:现代药理学研究表明,天麻的主要成分天麻素可以恢复大脑皮质兴奋与抑制过程间的平衡失调,产生镇静、安眠和镇痛等中枢抑制作用。目的:观察天麻素对L6大鼠成肌细胞化学损伤的保护作用。方法:用H2O2建立L6大鼠成肌细胞的化学损伤模型。DMEM培养液制备不同浓度的天麻素(1.562500,0.781250,0.390625g/L)预处理保护组的L6大鼠成肌细胞24h,然后用0.1mmol/LH2O2孵育80min,同时设置增殖组(用不同浓度的天麻素预处理,但不经H2O2处理),模型组(仅用H2O2处理),阳性对照组(仅有L6细胞悬液),阴性对照组(只有DMEM培养液),对照组既不用天麻素预处理,也不用H2O2孵育。结果与结论:MTT检测显示,增殖组和保护组L6大鼠成肌细胞存活率明显高于模型组;DAPI荧光标记细胞核为蓝色,模型组细胞荧光度较弱,且数目显著少于保护组;苏木精-伊红染色显示保护组的细胞核形状较规则,细胞轮廓较清晰,而模型组破损细胞较多;Bax和Bcl-2免疫荧光细胞化学结果表明,模型组促凋亡基因bax表达明显高于保护组,而抑制凋亡基因bcl-2的表达量完全相反;流式细胞仪检测显示保护组的凋亡率显著低于模型组(P<0.05)。提示天麻素对保护大鼠骨骼肌细胞化学损伤有显著效果。

Protection effect of Gastrodin on chemical damage of skeletal muscle cells in rats

BACKGROUND:Modern pharmacological studies have showed that Gastrodin can restore the imbalance between excitability and inhibition of cerebral cortex,produce sedation,sleep,analgesia and other central inhibition.OBJECTIVE:To investigate the protective effect of Gastrodin on chemical damage of skeletal muscle cells in L6 rats.METHODS:A chemical myoblast damage rat model of L6 was established by using H2O2.Samples of pre-protective group were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing different concentrations of Gastrodin (1.562 500,0.781 250,0.390 625 g/L) for a 24-hour pretreatment,followed by the incubation of H2O2 (0.1 mmol/L) for 80 minutes.At the same time,the proliferation group (different concentrations of Gastrodin for a 24-hour pretreatment,but not followed by the incubation of H2O2),model group (only the incubation of H2O2),positive control group (only L6 rat myoblast suspension),negative control group (only DMEM medium) and control group (without Gastrodin pretreatment and H2O2 incubation) were established.RESULTS AND CONCLUSION:3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the survival rate of the proliferation and pre-protective groups was significantly higher than that of the model group.Cell nuclei were labeled by DAPI turned blue,while the fluorescent intensity of DAPI in the model group was obviously much lower than that in the pre-protective group and so was the quantity of cells labeled by DAPI.Cell nuclei were more regularly shaped,more compact cytoplasm and clearer cell outline in the pre-protective group from hematoxylin-eosin staining.However,cells with structural damages were obviously observed in the model group.Bax and Bcl-2 immunofluorescent cytochemical technique demonstrated that the expression of bax in the model group was evidently higher than that in the pre-protective group,while the expression of bcl-2 was the exact opposite.According to the results of flow cytometry,the rate of apoptosis in the pre-protective group was significantly lower than that in the model group (P < 0.05).It is indicated that Gastrodin has remarkable effect on injury of skeletal muscle cells in rats induced by chemicals.