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脂质体包裹的相思子毒素诱导的致癌作用反应调节

Carcinogenesis response modulation induced by gelonin encapsulated in liposome.

作者信息

Alam Anis, Nakhuru K S, Singha L I

机构信息

Immunology Laboratory, Department of Biochemistry, North-Eastern Hill University, Shillong, Meghalaya, India.

出版信息

Mol Cell Biochem. 2008 Aug;315(1-2):85-95. doi: 10.1007/s11010-008-9792-7. Epub 2008 May 26.

Abstract

The effectiveness of gelonin to arrest protein synthesis, thereby limiting the growth of cancer cells was studied by encapsulating it into liposomes. The protein was extracted from the seeds of Indian plant Gelonium multiflorum by ammonium sulfate precipitation and purified using cation-exchange and gel-filtration chromatography. Biological activity of purified gelonin was determined using a rabbit reticulocyte lysate assay in the cell-free translational experiments. Gelonin was encapsulated in conventional liposomes prepared by the dry film method in order to retain biological activity of the entrapped protein. Carcinogenesis was induced in Swiss albino mice by intravenous administration of DBN (10 mg kg(-1) body weight) at weekly intervals. Marker enzyme assays (GGT, AChE, and GST), GSH levels, cell proliferation assay, hepatocyte DNA analysis, histological examination of micro sections of liver tissues were parameters used to monitor carcinogenesis induction, and regression in mice. From the in vitro experiments conducted, it was observed that gelonin upon its encapsulation into liposome, resulted in significant destruction of the transformed liver cells by its cytotoxic effects that arrest protein synthesis. Various parameters studied to monitor regression also suggested mass cell destruction to liver upon administration of liposomal gelonin in mice exposed to DBN.

摘要

通过将去甲核糖体毒素包裹在脂质体中来研究其抑制蛋白质合成从而限制癌细胞生长的有效性。该蛋白质通过硫酸铵沉淀从印度植物多花银胶菊种子中提取,并使用阳离子交换和凝胶过滤色谱法进行纯化。在无细胞翻译实验中,使用兔网织红细胞裂解物测定法测定纯化的去甲核糖体毒素的生物活性。为了保留包封蛋白质的生物活性,将去甲核糖体毒素包裹在通过干膜法制备的常规脂质体中。通过每周静脉注射二亚硝基苯(10 mg kg(-1)体重)诱导瑞士白化小鼠发生癌变。标记酶测定(γ-谷氨酰转肽酶、乙酰胆碱酯酶和谷胱甘肽S-转移酶)、谷胱甘肽水平、细胞增殖测定、肝细胞DNA分析、肝组织微切片的组织学检查是用于监测小鼠癌变诱导和消退的参数。从进行的体外实验中观察到,去甲核糖体毒素包裹在脂质体中后,通过其抑制蛋白质合成的细胞毒性作用导致转化的肝细胞显著破坏。在暴露于二亚硝基苯的小鼠中,研究的用于监测消退的各种参数也表明,给予脂质体去甲核糖体毒素后,肝脏出现大量细胞破坏。

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