刺血疗法对原发性肝癌大鼠VEGF、TGF-β1的影响

Effect of Blood Pricking Therapy on Serum VEGF and TGF-β1 in Rats with Hepatic Cellular Cancer

目的：观察刺血疗法对原发性肝癌（HCC）大鼠血管内皮生长因子（VEGF）、转化生长因子β1（TGF-β1）及肝脏病理形态学的影响,探讨其对HCC大鼠癌变的干预作用及其机制。方法：将40只雄性Wistar大鼠分为正常组（10只）、模型组（15只）、刺血预防组（15只）。采用腹腔注射二乙基亚硝胺（DEN）法诱导建立HCC模型,为期13周。正常组不进行任何干预;模型组只抓取按压;刺血预防组从造模开始即给予大鼠后肢肝经浅表静脉处刺血,每次0.3~0.5 ml,每周2次,至13周。第13周末,以10%水合氯醛麻醉大鼠,腹主动脉取血5 ml,采用ELISA法进行血清甲胎蛋白异质体（AFP-L3）和肿瘤血管生成相关因子VEGF、TGF-β1的检测,剖取肝叶行HE染色观察肝组织病理形态学改变。结果：与正常组相比,模型组大鼠AFP-L3、VEGF、TGF-β1含量均升高,差异具有统计学意义（P〈0.01）;与模型组相比,刺血预防组大鼠AFP-L3、VEGF、TGF-β1含量均降低,差异具有统计学意义（P〈0.05）。HE染色光镜下观察肝组织切片,模型组肿瘤灶诱导形成,癌结节较多;刺血预防组癌细胞数量较少,仅见少量结节形成。结论：刺血疗法具有预防或延缓肝癌形成的作用,其机制可能与降低肿瘤血管生长因子VEGF、TGF-β1的表达、抑制肿瘤血管生成有关。

Objective：To observe the effect of blood pricking therapy on vascular endothelial growth factor（ VEGF）,transforming growth factor-beta1（ TGF-β1） and liver pathological morphology in rats with hepatic cellular cancer（ HCC）,and to explore its interventive effect and mechanism on carcinogenesis of rats.Methods：A total of 40 male Wistar rats were randomly divided into three groups,that is,a normal group（ 10 rats）,a model group（ 15 rats） and a blood pricking prevention group（ 15 rats）.The HCC model was iuduced by intraperitoneal injection of diethylnitrosamine（ DEN） for 13 weeks.In normal group the rats received no treatment;in model group the rats were treated by fixation and pressure;from the beginning of the creation of the model,rats in the blood pricking preventing group were pricked for bloodletting 0.3 ~ 0.5 m L each time and 2 times a week on the superficial vein of the rats hind leg along the liver meridian until the thirteenth week.At the end of the thirteenth week,the rats were made anesthesia with 10% of chloral hydrate,then 5 m L blood was drawn from the abdominal aorta.Use the method of ELISA to detect the serum alpha-fetoprotein heteroplasmon-L3（ AFP-L3） and the tumor vessel angiogenesis related factors VEGF,TGF-β1.The liver with hepatic lobule was removed for HE staining so that the pathological morphological changes of liver tissue were observed.Results：Compared with the normal group,the levels of AFP-L3,VEGF and TGF-β1 content in the model group were significantly increased,and the difference had a statistical significance（ P 〈 0.01）;compared with the model group,the levels of AFP-L3,VEGF and TGF-β1 content in the blood pricking preventing group were significantly decreased,and the difference had a statistical significance（ P 〈 0.05）.In the observation of liver tissue sections under light microscope after HE staining,in the model group the rats had formd tumor and more cancer nodules,while in the blood pricking prevention group the rats had few cancer cells and nodules.Conclusion：Blood pricking therapy has an effect on preventing or delaying the formation of HCC,which may be related to the reduction of tumor vessel angiogenesis factors VEGF and TGF-β1 and to inhibiting the tumor vessel angiogenesis.