三七总皂苷对L_6大鼠成肌细胞抗化学损伤模型的保护作用

Protective effect of panax notoginseng saponin against chemical damage of myoblasts in L_6 rats

背景:三七总皂苷具有活血祛瘀,通脉活络等多重药理作用,但其具体机制尚不明确。目的:观察三七总皂苷对L6大鼠成肌细胞H2O2损伤的保护作用及其对Bax和Bcl-2蛋白表达的影响。方法:建立L6大鼠成肌细胞H2O2损伤模型,应用MTT法、DAPI化学荧光法和流式细胞仪分析检测各种剂量三七总皂苷(0.5,0.1,0.02g/L)对L6大鼠成肌细胞存活率的影响;通过免疫荧光抗体细胞化学法研究各种剂量三七总皂苷(0.5,0.1,0.02g/L)对L6大鼠成肌细胞Bax和Bcl-2蛋白表达的影响。结果与结论:L6大鼠成肌细胞经H2O2损伤后细胞存活率降低、凋亡率增加,三七总皂苷能明显提高经H2O2损伤后细胞的存活率,降低凋亡率;免疫荧光抗体细胞化学法表明三七总皂苷能促进Bcl-2表达,抑制Bax表达。结果可见三七总皂苷对L6大鼠成肌细胞H2O2损伤具有保护作用,依次为0.1g/L>0.02g/L>0.5g/L,其作用可能与上调Bcl-2蛋白的表达、抑制Bax蛋白的表达有关。

BACKGROUND：Panax notoginseng saponin has multiple pharmacological effects such as stasis and clearing and activating the channels and collaterals,but the mechanisms are unclear.OBJECTIVE：To observe the protective effect of panax notoginseng saponin against H2O2 damage of L6 rat myoblasts and its effect on the expression of Bax and bcl-2.METHODS：H2O2 damaged model of L6 rat myoblasts was built.3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay,4,6-diamino-2-phenyl indole chemical fluorescence and flow cytometry were applied to detect the survival rates of L6 rat myoblasts treated with different dosesof panax notoginseng saponin（0.5,0.1 and 0.02 g/L）.The effects of different doses of panax notoginseng saponin（0.5,0.1 and 0.02 g/L） on Bax and Bcl-2 protein expression levels of L6 rat myoblasts were analyzed by immunofluorescence cytochemistry.RESULTS AND CONCLUSION：The L6 rat myoblasts had low survival rate and high apoptosis rate after injury of H2O2,while panax notoginseng saponin could obviously increase the survival rate and reduce the apoptosis rate.Immunofluorescence cytochemistry results showed that panax notoginseng saponin could enhance the Bcl-2 expression and inhibit the Bax expression.Panax notoginseng saponin can protect L6 rat myoblasts from H2O2 damage,and the protective effects are in order of sequence of 0.1 g/L 0.02 g/L 0.5 g/L.The mechanism may be related to the enhancement of Bcl-2 expression and inhibition of Bax expression.