夏枯草通过调控OPG/RANK/RANKL信号通路对骨肉瘤小鼠的抑瘤作用研究

目的:探讨夏枯草通过调控OPG/RANK/RANKL信号通路对骨肉瘤小鼠的抑瘤作用。方法:60只昆明小鼠,其中10只为对照组,其余50只采用腋窝皮下注射0.1 mL的S180细胞法建立骨肉瘤小鼠模型,然后随机分为模型组,夏枯草低、高剂量(1 g·kg^-1、2 g·kg^-1)组,化疗(环磷酰胺(0.03 g·kg^-1)]组、夏枯草联合化疗(夏枯草高剂量(2 g·kg^-1)+环磷酰胺(0.03 g·kg^-1)]组。夏枯草组灌胃给药,对照组和模型组给予同体积的生理盐水,化疗组隔日腹腔注射环磷酰胺,均干预14 d。观察干预期间小鼠一般状况,末次给药后处死小鼠,对比各组瘤质量、抑瘤率;观察各组肿瘤组织病理学变化,并对比肿瘤细胞凋亡率、肿瘤组织中OPG、RANK、RANKL的基因和蛋白的表达。结果:建模期间小鼠一般状况及精神状态均随时间延长逐渐转差,各药物干预组干预期间有所好转,其中夏枯草联合化疗组改善最为明显。肿瘤组织病理学观察显示,模型组细胞排列紧密、细胞核大而深染、细胞膜完整,各干预组出现肿瘤细胞坏死、核浓缩或分裂、空泡等现象,其中夏枯草联合化疗组上述变化最明显。与模型组比较,其余4组瘤重明显减轻,RANK、RANKL的基因和蛋白表达量均明显降低(P<0.05),凋亡率、OPG和RNAKL的基因和蛋白表达量均明显升高(P<0.05)。结论:夏枯草对S180骨肉瘤小鼠有显著抑制作用,且具有增强化疗效果的作用,可能通过下调OPG/RANK/RANKL信号通路中RANK、RANKL表达,上调OPG表达及OPG/RANKL发挥作用。

Anti-tumor Effect of Xiaku Cao(Prunella Vulgaris)on Osteosarcoma Mice by Regulating OPG/RANK/RANKL Signaling Pathway

Objective:To investigate the anti-tumor effect of Xiaku Cao(Prunella Vulgaris)on osteosarcoma mice by regulating OPG/RANK/RANKL signaling pathway.Methods:Sixty SPF Kunming mice were selected,10 of which were in the control group,and the other 50 mice were injected subcutaneously with 0.1 mL S180 cells to establish osteosarcoma mouse model.They were randomly divided into model group,Xiaku Cao(Prunella Vulgaris)low,medium and high dose groups(1 g·kg^-1,2 g·kg^-1),chemotherapy group(cyclophosphamide(0.03 g·kg^-1)],Xiaku Cao(Prunella Vulgaris)combined with chemotherapy groupXiaku Cao(Prunella Vulgaris)high dose(2 g·kg^-1)+cyclophosphamide(0.03 g·kg^-1)].Xiaku Cao(Prunella Vulgaris)group was given medicine by gavage,control group and model group were given the same volume of normal saline,chemotherapy group was given intraperitoneal injection of cyclophosphamide every other day,and all of them were intervened for 14 days.The general condition of mice during the intervention period was observed,and the mice were killed after the last administration,and the tumor weight and tumor inhibition rate of each group were compared;the tumor histopathological changes of each group were observed,and the apoptosis rate of tumor cells,the gene and protein expression of OPG,RANK and RANKL in tumor tissue were compared.Results:During the modeling period,the general condition and mental state of the mice gradually deteriorated with the extension of time,and the drug intervention groups improved during the intervention period,especially the Xiaku Cao(Prunella Vulgaris)combined with chemotherapy group.Histopathological observation of tumor showed that the cells in the model group were closely arranged,the nuclei were large and deeply stained,and the cell membrane was intact.The tumor cells in the intervention groups were necrotic,nuclear concentration or division,vacuoles and other phenomena,and the above changes were most obvious in the Xiaku Cao(Prunella Vulgaris)combined with chemotherapy group.Compared with the model group,the tumor weight,the gene and protein expression of RANK and RANKL in the other four groups were significantly reduced(P<0.05),and the apoptosis rate,the gene and protein expression of OPG and OPG,RANKL were significantly increased(P<0.05).Conclusion:Xiaku Cao(Prunella Vulgaris)has a significant inhibitory effect on S180 osteosarcoma mice,and can enhance the effect of chemotherapy.It may play a role by down regulating the expression of rank and RANKL in OPG/RANK/RANKL signaling pathway,up regulating the expression of OPG and OPG/RANKL.