半枝莲多糖SBP-2A对小鼠肝癌H22细胞增殖的抑制作用

Inhibitory Effect of Scutellaria Barbata Polysaccharide SBP-2A on Proliferation of Mouse Hepatoma H22 Cells

目的探讨半枝莲多糖SBP-2A对小鼠肝癌H22细胞增殖及凋亡的影响。方法体外培养小鼠肝癌H22细胞分为阴性对照组(不加任何药物)、阳性对照组(黄芪多糖200μg/mL)、低剂量治疗组(半枝莲多糖SBP-2A 50μg/mL)、中剂量治疗组(半枝莲多糖SBP-2A 100μg/mL)、高剂量治疗组(半枝莲多糖SBP-2A 200μg/mL)。MTT比色法检测各组细胞增殖的抑制活性。HE染色观察各组细胞形态学变化。流式细胞术Annexin V/PI双染法检测各组细胞凋亡水平。蛋白质印迹法(Western blot,WB)检测各组细胞B细胞淋巴瘤2基因(B cell lymphoma 2 gene,Bcl-2)、BCL2关联X蛋白(BCL2-associated X protein,Bax)的蛋白表达水平。结果低、中、高剂量治疗组细胞增殖抑制率明显增加。低、中、高剂量治疗组均较阴性对照组出现一定程度的凋亡形态特征,且高剂量治疗组最为明显。高剂量治疗组细胞凋亡率显著高于阴性对照组及低、中剂量治疗组(P<0.05)。低、中、高剂量治疗组Bax蛋白表达水平均显著高于阴性对照组(P<0.05);中、高剂量治疗组Bcl-2蛋白表达水平均显著低于阴性对照组(P<0.05)。结论半枝莲多糖SBP-2A可显著抑制小鼠肝癌H22细胞增殖,其作用机制可能与调控Bax、Bcl-2蛋白表达水平从而促进细胞凋亡有关。

Objective To investigate the effect of Scutellaria barbata polysaccharide SBP-2A on the proliferation and apoptosis of mouse hepatoma H22 cells.Methods In vitro cultured mouse H22 cells were divided into negative control group(without any drug),positive control group(Astragalus polysaccharide 200μg/mL),low dose treatment group(Scutellaria barbata polysaccharide SBP-2A 50μg/mL),medium dose treatment group(Scutellaria barbata polysaccharide SBP-2A 100μg/mL),and high dose treatment group(Scutellaria barbata polysaccharide SBP-2A 200μg/mL).The proliferation inhibitory activity of cells in each groups was measured by MTT colorimetric method.Cell morphological changes in each group were observed by HE staining.The level of apoptosis in each group was determined by flow cytometry Annexin V/PI double staining.The protein expression levels of B cell lymphomas 2 gene(Bcl-2)and BCL2-associated X protein(Bax)were determined by Western blot(WB).Results Cell proliferation inhibition rate was significantly increased in the low,medium and high doses treatment groups.The low,medium and high doses treatment groups showed some degree of apoptotic morphology compared with the negative control group,and the most obvious is in the high dose treatment group.Cell apoptosis rate in the high dose treatment group was significantly higher than that in the negative control group and the low,medium doses treatment groups(P<0.05).Bax protein expression in the low,medium and high doses treatment groups was significantly higher than that in the negative control group(P<0.05),and Bcl-2 protein expression in the medium,high doses treatment groups was significantly lower than that in negative control group(P<0.05).Conclusion Scutellaria barbata polysaccharide SBP-2A could significantly inhibit mouse hepatoma H22 cell proliferation,and the mechanism of action might be related to the regulation of Bax and Bcl-2 protein expression levels and thus promote apoptosis.