摘要
目的探讨超声引导瘤内注射复方中药99-克星(99-克星)治疗裸鼠肝癌的作用机制、抗癌效果及其对机体的影响。方法选取人肝癌SMMC-7721皮下移植的实验裸鼠及肝内原位移植的实验裸鼠模型各14只,随机分为3组,其中99-克星治疗组14只,无水乙醇与生理盐水治疗组各7 只。全部裸鼠在接种10 d后每隔5 d向肿瘤内注入治疗药物共4次后处死,治疗前后分别采用高频超声测量肿瘤三径并计算肿瘤的生长指数。采用放射免疫法(RIA)测定治疗后荷瘤鼠血清白介素2 (IL-2)和肿瘤坏死因子(TNF-α)含量,采用末端脱氧核苷酸转移酶介导的dUTP切口末端标记法 (TUNEL)对细胞凋亡现象进行原位观察并观察血清TNF-α水平与肝癌细胞凋亡的关系。治疗前后分别测血天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)、尿素氮 (BUN)及白细胞计数。实验完成后解剖取检28只实验裸鼠心、肝、肾、脾、肺、脑等器官行病理形态学观察。实验均在双盲条件下进行。结果治疗后克星组肿瘤生长指数(0．080±0．024)明显低于盐水组(4．842±1．293)(P<0．01),并且低于乙醇组(0．083±0．026),但差异无统计学意义(P> 0．05);治疗后克星组血清IL-2和。TNF-α水平明显高于盐水组及乙醇组(P<0．05);克星组凋亡指数 (AI)高达(45．08±5．39)％,显著高于乙醇组(13．45±2．54)％和盐水组(12．38±3．91)％(P< 0．01);治疗前后ALT、AST、BUN及白细胞计数均无明显变化,3组间比较无统计学差异;血清TNF-α含量及肝癌细胞凋亡指数间呈正相关(r=0．494,P<0．05);3组实验裸鼠心、肝、肾、脾、肺、脑等器官均无病理形态学异常改变。结论 99-克星具有明显破坏和抑制肿瘤细胞生长的作用,而无抑制骨髓及损害肝肾功能等毒副作用,其作用机制不同于无水乙醇,主要通过提高血清细胞因子IL-2和TNF-α水平激发机体细胞免疫功能以及诱导癌细胞凋亡,从而达到抑制肿瘤生长的目的。
Objective To investigate the anti-cancer effect of ultrasound-guided intra-tumoral injection of traditional Chinese medicine ＂Star-99＂ in the treatment of hepatocellular carcinoma （ HCC ） and to determine whether ＂Star-99＂ has obvious adverse effects on the body. Methods Twenty-eight human hepatocellular carcinoma SMMC-7721 transplanted nude mice （including 14 hypodermically implanted mice and 14 orthotopic liver transplanted mice） were randomly divided into 3 groups ： Star-99 treatment group （ n = 14）, ethanol （ n = 7 ） and saline （ n = 7 ） treatment groups. Ten days after transplantation the study medicines were injected into the tumors of all the nude mice once every 5 days. After 4 injections the nude mice were killed. The 3 dimensions of the tumors were measured by high frequency ultrasound before and after treatment and the tumor growth indexes （TGI） were calculated [ （post-treatment tumor volume -pretreatment tumor volume） / pre-treatment tumor volume]. Radioimmunoassay （RIA） was used to detect the serum levels of interleukin-2 （IL-2） and tumor necrosis factor （TNF）-alpha. Apoptotic cells were evaluated by TUNEL assay. Liver and kidney function tests （AST, ALT, ALP and BUN） as well as white blood cell counts were determined before and after treatment. After experiment all the nude mice were investigated by autopsy and histological examination for the information of the heart, liver, kidney, spleen, lung and brain.All the experiments were carried out in double blind way. Results The TGI of Star-99 group （0. 080 ± 0. 024） was markedly lower than that of saline group （4. 842 ± 1. 293） （P 〈0.01）. It was also lower than that of ethanol group （0. 083 ± 0. 026） , but the difference was not significant （ P 〉 0.05 ）. Serum levels of IL-2 and TNF-α of Star-99 group were markedly higher than those of ethanol and saline groups （P 〈 0.05 ）. The mean apoptotic index （AI） in Star-99 group [ （45.08 ± 5.39） % ] was significantly higher than those of ethanol group [（13.45±2.54） %] and saline group [（12.38±3.91） %]（P〈0.01）. There were no evident changes of liver and kidney function as well as WBCs before and after treatment in the 3 groups. Positive correlation （ r = 0. 494, P 〈 0.05 ） was found between AI and serum level of TNF-α. Autopsy and histological examination of all the nude mice showed no evident changes of the heart, liver, kidney, spleen, lung and brain. Conclusions It is indicated that Star-99 can enhance the cellular immunity and induce cancer cell apoptosis. The major mechanisms can be as follows ： （ 1 ） it increases the serum levels of IL-2 and TNF-α and triggers cellular immunologic function; （2） by inducing cancer cells＇apoptosis, it can inhibit and destroy the cancer cells. The mechanism of action of Star-99 is different from that of ethanol. This study provides a new pathway of drug administration and an experimental basis for the treatment of HCC with Chinese herbal medicines, and it also provides an experimental basis for the ultrasound-guided intra-tumoral injection of Chinese medicine ＂Star-99＂ in clinical application.
出处
《中华医学超声杂志（电子版）》
2005年第3期131-135,共5页
Chinese Journal of Medical Ultrasound(Electronic Edition)
基金
福建省科学基金资助课题(NO:2000Z138)
