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肝脏S9酶生物转化对黑曲霉提取黑色素的致突变性和细胞毒性的影响。

Effect of biotransformation by liver S9 enzymes on the mutagenicity and cytotoxicity of melanin extracted from Aspergillus nidulans.

作者信息

de Cássia Ribeiro Gonçalves Rita, Rezende Kitagawa Rodrigo, Aparecida Varanda Eliana, Stella Gonçalves Raddi Maria, Andrea Leite Carla, Regina Pombeiro Sponchiado Sandra

机构信息

a Department of Pharmaceutical Sciences , Espirito Santo Federal University - UFES , Vitoria , Brazil .

b Faculty of Pharmaceutical Sciences , São Paulo State University - UNESP , Araraquara , Brazil , and.

出版信息

Pharm Biol. 2016;54(6):1014-21. doi: 10.3109/13880209.2015.1091846. Epub 2015 Oct 12.

Abstract

CONTEXT

A mutant that exhibited increased melanin pigment production was isolated from Aspergillus nidulans fungus. This pigment has aroused biotechnological interest due to its photoprotector and antioxidant properties. In a recent study, we showed that melanin from A. nidulans also inhibits NO and TNF-α production.

OBJECTIVE

The present study evaluates the mutagenicity and cytotoxicity of melanin extracted from A. nidulans after its exposure to liver S9 enzymes.

MATERIALS AND METHODS

The cytotoxicity of multiple concentrations of melanin (31.2-500 μg/mL) against the McCoy cell line was evaluated using the Neutral Red assay, after incubation for 24 h. Mutagenicity was assessed using the Ames test with the Salmonella typhimurium strains TA98, TA97a, TA100, and TA102 at concentrations ranging from 125 μg/plate to 1 mg/plate after incubation for 48 h.

RESULTS

The cytotoxicity of A. nidulans melanin after incubation with S9 enzymes was less than (CI50 value= 413.4 ± 3.1 μg/mL) that of other toxins, such as cyclophosphamide (CI50 value = 15 ± 1.2 μg/mL), suggesting that even the metabolised pigment does not cause significant damage to cellular components at concentrations up to 100 μg/mL. In addition, melanin did not exhibit mutagenic properties against the TA 97a, TA 98, TA 100, or TA 102 strains of S. typhimurium, as shown by a mutagenic index (MI)  <2 in all assays.

DISCUSSION AND CONCLUSION

The significance of these results supports the use of melanin as a therapeutic reagent because it possesses low cytotoxicity and mutagenic potential, even when processed through an external metabolising system.

摘要

背景

从构巢曲霉中分离出一种黑色素生成增加的突变体。这种色素因其光保护和抗氧化特性而引起了生物技术领域的关注。在最近的一项研究中,我们发现构巢曲霉黑色素还能抑制一氧化氮和肿瘤坏死因子-α的产生。

目的

本研究评估构巢曲霉黑色素经肝脏S9酶处理后的致突变性和细胞毒性。

材料与方法

使用中性红试验评估多种浓度(31.2 - 500μg/mL)的黑色素对 McCoy 细胞系的细胞毒性,孵育24小时后进行检测。使用鼠伤寒沙门氏菌菌株TA98、TA97a、TA100和TA102进行Ames试验评估致突变性,孵育48小时后,浓度范围为125μg/平板至1mg/平板。

结果

与S9酶孵育后的构巢曲霉黑色素的细胞毒性小于(CI50值 = 413.4 ± 3.1μg/mL)其他毒素,如环磷酰胺(CI50值 = 15 ± 1.2μg/mL),这表明即使是代谢后的色素在浓度高达100μg/mL时也不会对细胞成分造成显著损害。此外,黑色素对鼠伤寒沙门氏菌的TA 97a、TA 98、TA 100或TA 102菌株均未表现出致突变特性,所有试验中的致突变指数(MI)均<2。

讨论与结论

这些结果的意义支持将黑色素用作治疗试剂,因为即使通过外部代谢系统处理,它也具有低细胞毒性和致突变潜力。

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