肝动脉灌注碘油羟基磷灰石纳米粒对兔VX2肝肿瘤细胞凋亡、增生及血管形成的影响

The effects of lipiodol-hydroxyapatite nanoparticle on apoptosis,profiferation,and angiogenesis in hepatic tumor:experiment with rabbits

目的观察肝动脉灌注碘油羟基磷灰石纳米粒(nHAP)对兔 VX2肝肿瘤生长、坏死率、凋亡指数、微血管密度(MVD)及增殖细胞核抗原(PCNA)表达的影响,同时观察其对瘤兔生存期的影响。方法 80只新西兰白兔肝内肿瘤种植后2周,随机分为4组,每组20只,兔上腹正中开腹,胃十二指肠动脉插管固定,经肝动脉灌注给药,实验设生理盐水组(A 组)、单纯碘油组(B 组)、阿霉素碘油组(C 组)及碘油 nHAP 组(D 组)。治疗后1周及2周,采用 CT 检测肿瘤大小,计算肿瘤的生长率。治疗后2周,病理观察肿瘤区的坏死率,DNA 缺口末端标记(TUNEL)法检测肿瘤的凋亡指数,免疫组化方法测定肿瘤的 MVD 值。记录各组实验兔治疗后的存活期。结果治疗后1、2周,碘油nHAP 组肿瘤体积及生长率明显小于其他各组(均 P<0.05)。治疗后2周,碘油 nHAP 组肿瘤坏死率和凋亡指数大于其他各组(均 P<0.05)。单纯碘油组及阿霉素碘油组栓塞后,残余肿瘤区的 MVD(条)升高,两者分别为34±7和35±8,高于阴性对照 A 组(23±6,P<0.05);碘油 nHAP 组残余瘤区MVD(16±4)条低于其他组。栓塞组增殖指数低于阴性对照 A 组,碘油 nHAP 组增殖指数低于其他各组(均 P<0.05)。4组瘤兔治疗后的生存天数(d)分别为38±6、46±8、50±8、55±9。碘油 nHAP 组治疗后的生存期长于其他组(均 P<0.05)。结论肝动脉灌注碘油 nHAP 可明显抑制肿瘤生长,延长瘤兔的生存期,其可能通过促进肿瘤细胞坏死、诱导肿瘤细胞凋亡、抑制肿瘤细胞增殖和抑制肿瘤血管生成发挥抗肿瘤效应。

Objective To investigate the effects of lipiodol-hydroxyapatite nanopartich （lipi-nHAP） on the growth, necrosis, apoptosis, proliferation, and angiogenesis of hepatic tumor. Methods Ultrasound-emulsification was used to make lipi-nHAP Eighty New Zealand white rabbits underwent implantation of carcinoma ceils of the line VX2 into the left lobe of liver. Two weeks later the rabbits underwent catheterization into the gastroduedenal artery so that, and then the rabbits were randomly divided into four equal groups to receive infusion via the hepatic artery of different drugs： physiological saline （ Group A）, lipiedol （ Group B）, adriamycin ＋ lipiedol （ Group C）, and lipi-nHAP （ Group D）. Seven and 14 days after the treatment the size of tumor was observed by spiral CT scan, and the volume and growth rate of tumor were calculated. Two weeks after the treatment 8 rabbits from each group were kilted and their liver tumors were taken out and the survival rates of remaining rabbits were observed. The necrosis rate of the liver tumor was assessed by measuring the area of the tumor and the necrosis. The apoptotic rate was examined by TUNEL method. Mcrovessel density （MVD） was examined by immunohistochemistry anti-CD31 antibody. Anti-proliferating cell nuclear antigen （ PCNA ） monoclonal antibody was used to detect the expression of PCNA so as to calculate the proliferation index of the cells. Results The tumor volume and growth rate of Group D 7 and 14 days after treatment were both significantly lower than those of other groups （ all P 〈 0.05 ） and the necrosis rate and apoptotic index of Group D were both significantly higher than those of other groups （ all P 〈 0.05 ）. The values of MVD were higher in Groups C and D compared with those of Group A. Compared with those in other groups, the values of MVD and expression level of PCNA were significantly lower in group D （all P〈0.05）. The survival time of Group D was longer than those of other groups （all P 〈 0.05 ）. Conclusion lipi-nHAP Can suppress the growth of tumor, increase the tumor＇s necrosis rate and apoptotic index, inhibit the development of neovascularization, decrease the expression level of PCNA of residual tumor, and prolong the surviving time of the animals with hepatic tumor. It may become an effective embolization material to treat liver cancer.