夏枯草提取物诱导B、T淋巴瘤细胞凋亡的实验研究

目的:研究夏枯草提取物对人B淋巴瘤Raji细胞及T淋巴瘤Jurkat细胞增殖的影响及其差异,探讨夏枯草提取物体外抗淋巴瘤的作用机制。方法:不同浓度的夏枯草提取物分别作用于Raji细胞及Jurkat细胞;48 h后分别收集各组细胞,应用MTT法、琼脂糖凝胶电泳法及流式细胞技术分别检测每组细胞的增殖抑制率、细胞凋亡率及细胞凋亡情况;Western blot检测BCL-2及BAX蛋白表达变化。结果:不同浓度的夏枯草提取物对Raji及Jurkat细胞均有明显的增殖抑制作用(P0.01),且对Raji细胞的抑制作用大于Jurkat,其对Raji细胞IC50(18.01±0.92)μg/mL显著低于Jurkat细胞(25.47±0.96)μg/mL(P0.05);凝胶电泳检测结果表明夏枯草提取物处理Raji及Jurkat细胞后均出现凋亡相关DNA Ladder;随夏枯草提取物浓度的增加,Raji及Jurkat细胞的早期凋亡率均显著增加,与空白对照组比较差异显著(P0.01),浓度为15、20、25μg/mL的夏枯草提取物作用于Raji及Jurkat细胞后的早期凋亡率分别为(9.4±0.25)%、(21.68±0.46)%、(35.03±0.35)%和(4.06±0.14)%、(13.59±0.23)%、(22.92±0.20)%,同浓度条件下,Raji细胞的早期凋亡率显著高于Jurkat细胞(P0.01);随夏枯草提取物浓度增加,实验组细胞中BCL-2蛋白的表达逐渐减弱,BAX蛋白的表达逐渐增强,与空白对照组比较均有显著性差异(P0.05),同浓度条件下,Raji细胞内BCL-2蛋白表达的下降程度、BAX蛋白表达的增加程度显著高于Jur-kat细胞(P0.05)。结论:夏枯草提取物具有抑制淋巴瘤细胞增殖的作用,其机制可能与调节BCL-2和Bax蛋白的表达以诱导细胞凋亡有关,且夏枯草提取物对Raji细胞的抑制作用大于Jurkat细胞。

Experimental study of extract from Prunella vulgaris inducing B, T lymphoma cell apoptosis

Objective:To investigate the effects of extract from Prunella vulgaris on proliferation of human B lymphoma cell line Raji cells anf T lymphoma cell line Jurkat cells and discuss the mechanism.Methods:Used different concentrations of extract from Prunella vulgaris to treat Raji and Jurkat cells and collected cells after 48 h respectively,the proliferation inhibition rate,the DNA Ladder and the apoptosis rate of Raji and Jurkat cells were examined by MTT assay,agarose gel electrophoresis and flow cytometry respectively;Western blot was used to detect the change of BCL-2,BAX protein.Results:Different concentrations of the extract from Prunella vulgaris could inhibit the proliferation of both Raji and Jurkat cells remarkably(P<0.01),the IC50 of Raji cells,18.01±0.92 μg/mL,was lower than that of Jurkat,the difference was significant statistically(P<0.05);Apoptosis related DNA Ladder appeared after treated Raji and Jurkat cells with the extract from Prunella vulgaris;Compared with the control group,with the increase of the concentration of the extract from Prunella vulgaris,the early cell apoptosis rate of Raji and Jurkat were all increased,the early cell apoptosis rate of the extract from Prunella vulgaris of 15,20 and 25 μg/mL treated Raji and Jurkat cells were(9.46±0.25)%,(21.68±0.46)%,(35.03±0.35)% and(4.06±0.14)%,(13.59±0.23)%,(22.92±0.20)% respectively.With the same concentration,the early apoptosis rate of Raji cells was higher than that of Jurkat cells significantly(P<0.01);Compared with the control group,with the increase of the concentration of the extract from Prunella vulgaris,the expression of BCL-2 protein was down-regulated and BAX up-regulated,With the same concentration,the decline degree of BCL-2 protein expression and the increase degree of BAX protein expression in Raji cells was more remarkable than that in Jurkat cells,the difference was significant(P<0.05) Conclusion:The extract from Prunella vulgaris can inhibit the proliferation of lymphoma cells and the inhibition is realized by inducing apoptosis,the mechanism of inducing cell apoptosis with extract from Prunella vulgaris is probably related with the BAX and BCL-2 protein expression,the inhibition effect on Raji cells is greater than that of Jurkat cells.