加味当归贝母苦参丸对MFC胃癌荷瘤小鼠瘤组织Toll样受体通路分子的影响

Effects of Modified Danggui Beimu Kushen Pills on Toll-like Receptor Pathway Markers on MFC Gastric Cancer Mice

目的观察加味当归贝母苦参丸对MFC胃癌荷瘤小鼠瘤组织Toll样受体(TLR2、TLR4、TLR6)、肿瘤坏死因子受体相关因子6(TRAF6)和髓分化因子88(My D88)表达的影响,探讨其相关的作用机制。方法建立MFC胃癌荷瘤小鼠模型进行体内抗肿瘤试验。将筛选合格的48只MFC胃癌荷瘤小鼠随机分为模型组,阳性对照组(顺铂组),加味当归贝母苦参丸高剂量组(当高组)、低剂量组(当低组)和顺铂联合加味当归贝母苦参丸高剂量组(顺高组)、低剂量组(顺低组),各给药组给予相应药物灌胃,连续14 d,观察肿瘤生长情况及小鼠一般情况、肿瘤大体标本及HE染色后组织形态;RT-q PCR和免疫组化分别检测肿瘤组织TLR2、TLR4、TLR6、TRAF6和My D88的表达。结果模型组肿瘤细胞丰富,细胞大小不一、体积大、核大深染、异型明显,可见小灶性坏死,间质血管丰富;各给药组肿瘤细胞数量减少、排列疏松、细胞体积明显缩小、核固缩,细胞坏死明显增加,间质血管数量减少,胶原纤维增多,其中以顺低组、顺高组较为明显。与模型组比较,各给药组TLR2、TLR4、TLR6、TRAF6和My D88 m RNA和蛋白表达均降低,顺低组、顺高组TLR2、TLR4、TLR6、TRAF6和My D88蛋白着色变浅,呈弱阳性表达,较顺铂组、当高组、当低组蛋白改变更加明显。结论加味当归贝母苦参丸可能通过下调荷瘤小鼠肿瘤组织TLR2、TLR4、TLR6、TRAF6和My D88 m RNA和蛋白的表达,发挥抗肿瘤作用。

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2,TLR4,TLR6,TRAF6and MyD88in tumor tissues on MFC gastric cancer bearing mice;To discuss relevant mechanism of action.Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study.A total of eligible48mice were randomly divided into model group,DDP positive control group,modified Danggui Beimu Kushen Pills high-dose and low-dose groups,modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups.The treatment was conducted once a day,and lasted for14continuous days.After the last administration of gavage orally treatment,all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2,TLR4,TLR6,TRAF6and MyD88in tumor tissue with the method of RT-qPCR and immunohistochemistry.Meanwhile,the tumor growth was observed and the general conditions of mice were recorded.Results The model group was rich in tumor cells;the sizes of cells were different;the volume was large;the nucleus was deeply stained and the heterotypic shape was obvious,and the small focal necrosis was seen.The number of tumor cells in each administration group was reduced;the arrangement was loose;the cell volume was significantly reduced,and the nuclear pyknosis was reduced.Cell necrosis significantly increased;the number of interstitial blood vessels decreased;collagen fibers increased,especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups.Compared with the model group,the expressions of TLR2,TLR4,TLR6,TRAF6and MyD88mRNA and protein decreased in each administration group.TLR2,TLR4,TLR6,TRAF6and MyD88were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups,the protein changes were more obvious Compared with DDP positive control group,modified Danggui Beimu Kushen Pills high-dose and low-dose groups.Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2,TLR4,TLR6,TRAF6and MyD88expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.