RGD肽段连接的近红外量子点对小鼠的毒性作用

doi:10.13481/j.1671-587x.20150404

摘要/Abstract

摘要：

目的: 探讨近红外量子点(NIR QDs)与含有精氨酸-甘氨酸-天冬氨酸序列(RGD)肽段连接的NIR QDs-RGD探针对小鼠的毒性作用,阐明NIR QDs-RGD作为一种新型肿瘤治疗手段的生物安全性。方法: 用NIR-QDS与RGD肽段连接,制备探针;将24只BALB/C小鼠随机分为4组,单次注射组在第1 天通过尾静脉注射100 μL的QD800-RGD(含200 pmol的QD800);单次注射对照组在第1天通过尾静脉注射100 μL的PBS;重复注射组分别在第1 天和第7天通过尾静脉各注射100 μL的QD800-RGD(含200 pmol的QD800);重复注射对照组分别在第1天和第7 天通过尾静脉各注射100 μL的PBS。注射14 d后行毒性检测,包括小鼠全血细胞(白细胞、红细胞、血红蛋白、血小板、淋巴细胞和中性粒细胞)计数和血清生化分析(总蛋白、白蛋白、白球比、天门冬氨酸氨基转换酶、丙氨酸氨基转换酶和血尿素氮),计算小鼠肝、脾、肾和肺的脏器系数,检测肝、脾、肾和肺组织中超氧化物歧化酶(SOD)、谷胱甘肽(GSH)活性和丙二醛(MDA)含量,并进行组织病理学检查。结果: 与各自对照组比较,单次注射组小鼠脏器系数稍有降低,重复注射组小鼠脏器系数增高,但4组小鼠之间比较差异无统计学意义(P>0.05)。全血细胞计数,与对照组比较,单次注射组小鼠白细胞、红细胞、血红蛋白、血小板、淋巴细胞数都有所增高,中性粒细胞数未见明显变化;重复注射组小鼠红细胞、血红蛋白、淋巴细胞增高,白细胞、血小板减少,中性粒细胞无明显变化;4组之间比较差异无统计学意义(P>0.05)。血清生化指标检测,与对照组比较,单次注射组小鼠总蛋白、白蛋白、白球比、血尿素氮减少,天门冬氨酸氨基转换酶和丙氨酸氨基转换酶增加;重复注射组小鼠总蛋白、白蛋白增加,白球比、血尿素氮减少,天门冬氨酸氨基转换酶和丙氨酸氨基转换酶降低;4组之间比较差异无统计学意义(P>0.05)。氧化和抗氧化指标的检测,与对照组比较,单次注射组和重复注射组小鼠肝、脾、肾和肺组织中SOD、GSH活性、MDA含量有少量增加,但4组之间比较差异无统计学意义(P>0.05)。肝、脾、肾和肺组织病理学检查,与对照组比较,单次注射组和重复注射组小鼠肝、脾、肾和肺的细胞形态规则,分布排列有序,均无坏死。 结论: NIR QD800-RGD在能达到对体内肿瘤成像所需的剂量下重复静脉注射在小鼠体内未导致明显的毒性作用。

关键词: 近红外量子点, RGD肽段, 毒性

Abstract:

Objective To study the toxicity of near-infrared quantum dots (NIR QDs) linked to peptide containing the arginine-glycine-aspartic acid (RGD) sequence (NIR QDs-RGD) in the mice,and to clarify the biological safety of NIR QDs-RGD as a novel tumor treatment method. Methods The probe was prepared firstly by linking NIR QDs to the RGD peptide.Twenty-four BALB/C mice were randomly divided into four groups.In QD800-RGD one injection group,the mice were injected via tail vein with 100 μL QD800-RGD (equivalent to 200 pmol of QD800) on the first day;in PBS one injection group,the mice were injected similarly with 100 μL PBS;in QD800-RGD two injection group,the mice were repeatedly injected via tail vein with 100 μL QD800-RGD (equivalent to 200 pmol of QD800) on the first and seventh days;in PBS two injection group,the mice were repeatedly injected similarly with 100 μL PBS on the first and seventh days.Fourteen days after injection,toxicity tests were performed,including complete blood cells(white blood cells,red blood cells,hemoglobin,platelets,lymphocytes,and neutrophils) count and serum biochemical analysis (total protein,albumin,albumin/globulin,aspartate aminotransferase,alanine aminotransferase,and blood urea nitrogen).The coefficients of liver,spleen,kidney,and lung weight to body weight were calculated,and the activities of superoxide dismutase(SOD),glutathione(GSH),and malondialdehyde(MDA) contents in liver,spleen,kidney and lung tissues of the mice were detected.The histopathological examination was performed. Results Compared with PBS injcetion groups,the organ coefficients of the mice in QD800-RGD one injection group were decreased slightly,and the organ coefficients of the mice in QD800-RGD two injection group were increased,but the differences between four groups were not statistically significant (P>0.05).Compared with PBS injcetion groups,the number of leukocytes,erythrocytes,lymphocytes and the levels of hemoglobin and platelets were increased,and the number of neutrophils had no significant change;the number of red blood cells,lymphocytes and hemoglobin level in QD800-RGD two injection group were increased;the number of white blood cells and the level of platelets were decreased,and the number of neutrophils had no significant changes;but the differences were not statistically significant between four groups (P>0.05).Compared with PBS injection group,the total protein,albumin,albumin globulin ratio,and blood urea nitrogen in QD800-RGD one injection group were reduced;the aspartate aminotransferase and alanine aminotransferase were increased;the total protein and albumin were increased,the albumin globulin ratio,blood urea nitrogen,aspartate aminotransferase and alanine aminotransferase in QD800-GRD two injection group were decreased;but the differences were not statistically significant between four groups (P>0.05). Compared with PBS injection group,the activities of SOD,GSH and the MDA contents in liver,spleen,kidney and lung tissues in QD800-RGD one injection group and QD800-RGD two injection group were slightly increased;but the differences were not statistically significant between four groups (P>0.05). Compared with PBS injection group,the morphology of liver,spleen,kidney and lung tissues in QD800-RGD one injection group and QD800-RGD two injection group were normal,the cells agganged orderly, and no necrosis was seen. Conclusion At the dose needed to produce in vivo tumor imaging,there is no significant toxicity of NIR QD800-RGD in the mice after repeatedly intravenous injection.

Key words: near-infrared quantum dots, RGD peptide, toxicity

中图分类号:

R979.1

陈睿, 李雯娟, 杨凯, 王有为, 唐洪, 陈丹, 白云龙, 黄昊. RGD肽段连接的近红外量子点对小鼠的毒性作用[J]. 吉林大学学报(医学版), 2015, 41(04): 690-696.

CHEN Rui, LI Wenjuan, YANG Kai, WANG Youwei, TANG Hong, CHEN Dan, BAI Yunlong, HUANG Hao. Toxicity of RGD peptide conjugated near-infrared quantum dots in mice[J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 690-696.

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