Biochemistry Division, Chemistry Department, Faculty of Science, Tanta University, Tanta, Egypt.
Zoology Department, Faculty of Science, Tanta University, Tanta, Egypt.
Environ Sci Pollut Res Int. 2020 Jun;27(16):19016-19028. doi: 10.1007/s11356-018-3339-6. Epub 2018 Oct 6.
This study was conducted to evaluate the effect of chitosan nanoparticles (CNPs) isolated from Artemia salina against hepatocellular carcinoma (HCC) both in vitro (HepG2) and in vivo (diethylnitrosamine-induced HCC in rats) and to investigate the involved underlying mechanisms. Administration of CNPs decreased HCC progression as evidenced by (1) induced HepG2 cell death as detected by MTT assay; (2) induced necrosis as indicated by acridine orange/propidium iodide (AO/PI) red staining, annexin V/7-AAD positive staining (detected by flow cytometry), and upregulated expression of necrosis markers (PARP1 and its downstream target, RIP1 genes), but no effect on apoptosis as revealed by insignificant changes in caspase 3 activity and mRNA levels of Bax and AIF; (3) increased intracellular ROS and decreased mitochondrial membrane potential in HepG2; (4) decreased liver relative weight, serum levels of liver enzymes (ALT, AST, and ALP), total bilirubin, and cancer markers (AFP and GGT), number and area of GST-P positive tumor nodules; and (5) reduced oxidative stress (decrease in MDA levels) and increased activities of SOD, CAT, and GPx enzymes in rat liver. The preventive (pre-treatment) effect of CNPs was better than the therapeutic (post-treatment) effect. Collectively, administration of CNPs inhibited HCC progression in vitro and in vivo, possibly through induction of necrosis, rather than apoptosis, and induction of antioxidant enzyme activities in vivo, but with stimulation of ROS production in vitro. Thus, CNPs could be used as a promise agent for treating HCC after application of further confirmatory clinical trials.
本研究旨在评估从卤虫中分离得到的壳聚糖纳米粒子（CNPs）对体外（HepG2）和体内（二乙基亚硝胺诱导的大鼠肝癌）肝癌的作用，并探讨其潜在的作用机制。CNPs 的给药降低了 HCC 的进展，这表现在：（1）MTT 检测到诱导 HepG2 细胞死亡；（2）吖啶橙/碘化丙啶（AO/PI）红色染色、膜联蛋白 V/7-AAD 阳性染色（流式细胞术检测）和坏死标志物（PARP1 及其下游靶标 RIP1 基因）的上调表达，表明诱导了坏死，但对凋亡没有影响，因为 caspase 3 活性和 Bax 和 AIF 的 mRNA 水平没有显著变化；（3）增加 HepG2 中的细胞内 ROS 和降低线粒体膜电位；（4）降低肝相对重量、血清肝酶（ALT、AST 和 ALP）、总胆红素和肿瘤标志物（AFP 和 GGT）水平、GST-P 阳性肿瘤结节的数量和面积；（5）降低 MDA 水平并增加大鼠肝中 SOD、CAT 和 GPx 酶的活性。CNPs 的预防（预处理）作用优于治疗（后处理）作用。总之，CNPs 的给药抑制了体外和体内 HCC 的进展，可能是通过诱导细胞坏死而不是凋亡，以及诱导体内抗氧化酶活性，但在体外刺激 ROS 产生。因此，CNPs 可在进一步进行临床试验证实后，用于治疗 HCC。
