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目的探讨不同光照剂量的光动力疗法(PDT)对小鼠Heps肝癌移植瘤的抗肿瘤作用及局部免疫效应。方法 69只ICR小鼠皮下接种Heps肝癌细胞建立荷瘤鼠模型,随机分为对照组、光照低剂量PDT组(能量密度135 J/cm2)和光照高剂量PDT组(能量密度215~225 J/cm2)。治疗后定期测量各组肿瘤体积,计算抑瘤率。观察HE染色的瘤组织病理学改变,免疫组化法检测肿瘤组织的CD8+细胞毒性T淋巴细胞(CD8+CTLs)。结果 (1)光照低剂量PDT组和光照高剂量PDT组抑瘤率分别为73.9%、96.3%;光照高剂量PDT组的抑瘤效果显著高于光照低剂量PDT组(P<0.01)。(2)PDT后2、24和72 h肿瘤组织先后出现肿瘤细胞空泡样变性、广泛核固缩及凝固性坏死;肿瘤中微血管淤血扩张、出血和管壁破坏,上述病理改变光照高剂量PDT组更为显著。(3)治疗后72 h光照低剂量PDT组和光照高剂量PDT组肿瘤中CD8+CTLs数量均明显高于对照组(P<0.01,P<0.05);而光照低剂量PDT组肿瘤中CD8+CTLs数量又显著高于光照高剂量PDT组(P<0.05)。结论光照高剂量PDT组的抑瘤效果显著高于光照低能剂量PDT组;PDT可引起细胞毒性T淋巴细胞在肿瘤局部浸润增加,光照低剂量PDT组对肿瘤局部CD8+CTLs的免疫增强效应优于光照高剂量PDT组。
Objective To investigate the anti-tumor effect and local immunologic effect of photodynamic therapy (PDT) with different light doses on Heps hepatocellular carcinoma in mice. Methods Sixty nine ICR mice were inoculated subcutaneously with Heps liver cancer cells to establish a tumor-bearing mouse model. The mice were randomly divided into control group, low dose PDT group (energy density 135 J / cm2) and high dose PDT group (energy density 215-225 J / cm2). After treatment, the tumor volume of each group was measured regularly and the inhibition rate was calculated. The histopathological changes of HE staining were observed. The CD8 + cytotoxic T lymphocytes (CD8 + CTLs) in tumor tissue were detected by immunohistochemistry. Results (1) The inhibition rates of low dose PDT group and high dose PDT group were 73.9% and 96.3%, respectively. The inhibition effect of high dose PDT group was significantly higher than that of low dose PDT group (P <0.01). (2) At 2, 24, and 72 h after PDT, tumor cells were vacuolizing degeneration, extensive nuclear pyknosis and coagulation necrosis. Tumor microvascular congestion expansion, hemorrhage and wall destruction. The above pathological changes of high dose PDT Group more significant. (3) The numbers of CD8 + CTLs in low dose PDT group and high dose PDT group at 72 h after irradiation were significantly higher than those in control group (P <0.01, P <0.05) The number of CTLs was significantly higher than that of high dose PDT group (P <0.05). Conclusions The tumor-inhibiting effect of high-dose PDT group is significantly higher than that of low-dose PDT group. PDT can increase the local infiltration of cytotoxic T lymphocytes in tumor, and enhance the immune enhancement effect of low dose PDT group on local CD8 + CTLs Light high dose PDT group.