电针联合扶正理气合剂对大鼠肝癌生长及转移的影响及相关机制

Effects of electroacupuncture combined with Fuzheng-Liqi mixture on liver cancer growth and metastasis in rats and the related mechanisms

目的观察电针联合扶正理气合剂对大鼠肝癌生长及转移的影响并探讨其作用机制。方法雄性Wistar大鼠200只,随机分为5组。以二乙基亚硝胺(DEN)灌胃诱导大鼠肝癌模型,电针组造模同时给予电针足三里、关元、内关、三阴交、肝俞穴治疗,扶正理气合剂组给予扶正理气合剂灌胃,针药联合组则予上述电针和扶正理气合剂治疗,共16周。造模16周后处死大鼠取肝脏组织标本,肉眼及光镜下观察肿瘤生长和转移情况;免疫组化法测定肝组织NF-κB活性及微血管密度(MVD),RT-PCR法检测肝组织中转化生长因子β1(TGF-β1)mRNA及血管内皮生长因子(VEGF)mRNA表达,放射免疫法测定肝组织活性氧(ROS)、抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性、脂质过氧化产物(LPO)含量。结果与正常对照组比较,模型组大鼠的肿瘤生长和转移参数、ROS、LPO含量、NF-κB活性、TGF-β1 mRNA、VEGF mRNA、MVD表达显著增加(P<0.01),而T-AOC、SOD、GSH-Px、CAT活性明显下降(P<0.01);与模型组比较,各治疗组大鼠的肿瘤生长和转移参数、ROS、LPO含量、NF-κB活性、TGF-β1mRNA、VEGF mRNA及MVD表达显著下降,而T-AOC、SOD、GSH-Px、CAT活性明显上升(P<0.01),联合治疗组上述指标优于其他治疗组(P<0.05)。NF-κB活性与TGF-β1mRNA及VEGF mRNA呈正相关关系(r1=0.554,P<0.05;r2=0.572,P<0.05)。结论电针和扶正理气合剂均能显著降低实验性肝癌大鼠的NF-κB活性、TGF-β1mRNA及VEGF mRNA及MVD表达,从而降低肿瘤生长和转移参数,这与其清除自由基有密切关系。

Objective To observe the effects of electroacupuncture combined with Fuzheng-Liqi mixture on growth and metastasis of rat liver cancer and to explore the related mechanisms.Methods Totally 200 male Wistar rats were randomly divided into five groups.Liver cancer model was induced with intragastric administration of diethylnitrosamine(DEN).At the same time of modeling,each rat in electroacupuncture group was treated with electroacupuncture of points Zusanli,Guanyuan,Neiguan,Sanyinjiao and Ganyu while Fuzheng-Liqi mixture was poured into the gastric cavity in Fuzheng-Liqi mixture group,and each rat was treated with the above-mentioned puncture and Fuzheng-Liqi mixture.After 16 weeks the rats were killed to check tumor growth and metastasis with the naked eye and light microscope.NF-κB activity and microvascular density(MVD) in liver tissues were determined with immunohistochemical method;TGF-β1mRNA and VEGFmRNA expressions were measured with RT-PCR method.And the levels of ROS,T-AOC,SOD,GSH-Px,CAT and LPO in liver tissues were determined with radioimmunoassay.Results Compared with those in normal control group,tumor growth and metastasis parameters,ROS,LPO,NF-κB activity,TGF-β1 mRNA,VEGF mRNA and MVD expressions were increased obviously while the levels of T-AOC,SOD,GSH-Px and CAT were decreased obviously in model group(P<0.01); Compared with those in model control group,tumor growth and metastasis parameters,ROS,LPO,NF-κB activity,TGF-β1 mRNA,VEGF mRNA and MVD expressions were decreased obviously while the levels of T-AOC,SOD,GSH-Px and CAT were increased obviously in all the treatment groups(P<0.01).The above-mentioned indexes were better in combination group than in other treatment groups(P<0.05).NF-B activity had an obvious positive correlation with TGF-β1 mRNA and VEGFmRNA expressions(r1=0.554,P<0.05;r2=0.572,P<0.05). Conclusion Electroacupuncture and Fuzheng-Liqi mixture can obviously decrease NF-κB activity,TGF-β1 mRNA,VEGFmRNA and MVD expressions in rats with hepatic tumor,thus decreasing tumor growth and metastasis parameters.This is closely correlated to their scavenging effects on free radicals.