张仲景抵当汤抑制肝癌门静脉瘤栓形成的组织学证据研究

Histologic evidence of inhibition of portal vein tumor thrombosis iinr hepatocellular carcinoma by Zhang Zhongjing's Didang Decoction

目的探寻抵当汤介导肝癌侵袭、转移及门静脉瘤栓形成的组织学证据。方法根据文献报道及既往研究结果,分别选取高侵袭性的肝癌细胞株HepG2和HCCLM3两种人肝癌细胞;调整细胞浓度,行裸鼠下腹腔注射3x10°个癌细胞(均300μL),以建立肝癌门静脉瘤栓模型。模型鉴定并复制,随机分为4组,分别为HepG2、HCCLM3生理盐水组(对照组),HepC2.HCCLM3抵挡汤组,每组6只,于腹腔注射癌细胞后第2周开始灌胃给药,对照组予同等量生理盐水,连续5周。比较四组裸鼠一般状态.网膜和肝脏成瘤率、血管侵犯和门静脉微瘤栓形成情况,采用免疫组化观察血管内皮生长因子(vascular.endothelial growth fator,VEGF)、表皮生长因子受体(Epidermal growth factor receptor,EGFR)表达情况。结果裸鼠腹腔接种初期一般状态无明显改变.至给药第5周时,生理盐水组2种肝癌细胞网膜成瘤率、肝脏成瘤率相当,HCCLM3细胞网膜血管侵犯比例高于HepG2,P<0.05,HCCLM3细胞门静脉微瘤栓发生率显著高于HepG2细胞,P<0.05。抵当汤组HCCLM3细胞网膜成瘤率、肝脏成瘤率、血管侵犯、微瘤栓及VEGFEGFR表达显著少于生理盐水组,P<0.05,抵当汤组HepG2细胞网膜成瘤率亦减少。结论抵当汤能明显抑制肝癌细胞的肝脏转移及门静脉瘤栓形成,以其破血逐瘀之峻效,抑制肿瘤血管新生,或可使肿瘤微小转移灶处于长期休眠状态。

Objective To explore the histological evidence of Didang Decoction mediating hepatocellular carcinoma invasion,metastasis and portal vein tumor thrombosis.Methods According to the literature reports and previous studies,two highly invasive hepatocellular carcinoma cell lines,HepG2 and HCCLM3,human hepatocellular carcinoma cells were selected respectively;the cell con-centration was adjusted,and the nude mice were injected with 3×10°cancer cells(both 300μL)in the lower peritoneal cavity,in order to establish the hepatocellular carcinoma portal vein tumor thrombus model.The model was identified and replicated,and randomly di-vided into 4 groups,namely,HepG2 and HCCLM3 saline group(control group),HepG2 and HCCLM3 Didang Decoction group,6 mice in each group,and started to be administered by gastric gavage in the 2nd week after intraperitoneal injection of cancer cells,and the control group was given the same amount of saline,for 5 consecutive weeks.The four groups of nude mice were compared in terms of general status,tumorigenicity of the omentum and liver,vascular invasion and portal vein microtumor thrombosis,and immunohistochem-istry was used to observe the expression of vascular endothelial growth factor(VECF)and epidermal growth factor receptor(EGFR).Results The general state of nude rats did not change significantly at the early stage of intraperitoneal inoculation,and by the 5th week of drug administration,the rates of retinal tumorigenesis and liver tumorigenesis of the two types of hepatocellular carcinoma cells in the saline group were comparable,and the proportion of retinal vascular invasion of HCCLM3 cells was higher than that of HepG2 cells,P<0.05,and the incidence of portal microtumor thrombosis of HCCLM3 cells was significantly higher than that of HepG2 cells,P<0.05.The rate of retinal tumorigenesis,liver tumorigenesis and retinal vascular invasion of HCCLM3 cells of the Didang Decoction group were similar to those of the saline group.The rate of retinal tumorigenesis,hepatic tumorigenesis,vascular invasion,microtumor thrombus and VECF,ECFR expression in HCCLM3 cells in the Didang Decoction group was significantly lower than that in the saline group,P<0.05,and the rate of retinal morigenesis in HepG2 cells in the Didang Decoction group was also reduced.Conclusion Didang Decoction can significantly inhibit liver metastasis and portal vein thrombus formation of epatocellular carcinoma cells,and inhibit tumor neovascularization by its acute effect of breaking blood and expelling blood stasis,which may make tumor micro-metastatic foci dormant for a long period of time.