耐辐射奇球菌R1对大鼠的慢性毒性实验

doi:10.12300/j.issn.1674-5817.2020.213

摘要/Abstract

摘要： 目的观察耐辐射奇球菌R1（Deinococcus radiodurans R1,DRR1）对SD大鼠的慢性毒性。方法将高浓度（10⁹/mL,LDRH组）和低浓度（10⁷/mL,LDRL组）的DRR1活菌以及高浓度（10⁹/mL,BDRH组）和低浓度（10⁷/mL,BDRL组）的细菌破碎物以1 mL/100 g体质量的剂量灌胃SD大鼠,同时以灌胃相同体积的TGY琼脂培养液（含0.5%胰蛋白胨、0.1%葡萄糖和0.3%酵母提取物）作为对照组（TGY组）;每组16只,雌雄各半,每日灌胃给药1次,连续30 d。每组随机取8只大鼠,用3%戊巴比妥钠0.15 mL/100 g体质量腹腔注射麻醉,收集血液及脏器标本,检测血常规、血清电解质、血清生化指标和脏器系数;主要脏器经体积分数为10%的中性甲醛溶液固定,行HE染色,光学显微镜下观察病理学变化。每组其余8只大鼠于停药14 d后作相同处理,观察DRR1的延迟毒性。实验期内观测大鼠行为、摄食量和体质量等一般情况。结果不同浓度的DRR1活菌及DRR1破碎物给药30 d后,停药即刻及停药14 d后,未见各组大鼠的运动、呼吸异常,未观察到反应迟钝、瞳孔改变、眼球凸出、皮肤颜色改变等异常现象,口、耳、鼻、眼角均未见异常分泌物,排便及形态正常;摄食量、体质量变化和脏器系数与TGY组比较,均无显著改变;白细胞数在给药30 d时显著下降（P<0.05）,停药14 d后恢复至正常水平;红细胞数、红细胞压积和血红蛋白含量无显著变化,淋巴细胞、粒细胞、单核细胞数及其百分比无显著变化,血小板数也无显著变化。停药后即刻血清谷丙转氨酶（ALT）、碱性磷酸酶（ALP）、总蛋白（TP）、尿酸（UA）、尿素（UREA）、血糖（Glu）、镁（Mg）和磷（P）含量无显著变化,血清谷草转氨酶（AST）、肌酸激酶（CK）、肌酸激酶同工酶（CK-MB）和乳酸脱氢酶（LDH）在LDRH组显著升高（P<0.05）;经14 d恢复期后,AST、CK降至恢复期TGY组水平,CK-MB仍高于恢复期TGY组（P<0.05）;恢复期的LDH水平与停药即刻比较,显著增高（P<0.05）,与恢复期TGY组比较无显著差异。与TGY对照组比较,给药30 d时血清离子浓度在各给药组间没有显著变化（P>0.05）;停药14 d后,LDRH组血清Cl^-、Ca²⁺降低,Na⁺、pH值升高（P<0.05）;对心、肝、肺和肾的病理学观察均未见明显实质性病变。结论 DRR1活菌及其细菌破碎物对大鼠无实际毒性,该实验为DRR1的拓展应用提供了实验数据。

关键词: 耐辐射奇球菌, 摄食量, 细菌破碎物, 慢性毒性, SD大鼠

Abstract: Objective To observe the chronic toxicity of Deinococcus radiodurans R1 (DRR1) in SD rats.Methods SD rats were divided randomly into 5 groups: 10⁹/mL live DRR1 group (LDRH), 10⁷/mL live DRR1 group (LDRL), 10⁹/mL DRR1 broken products group (BDRH), 10⁷/mL DRR1 broken products group (BDRL), and agar medium including 0.5% tryptone, 0.1% glucose and 0.3% yeast agar medium as control group (TGY), and 16 rats in each group with half males and half females. The rats were gavaged once a day for consecutive 30 days,and then 8 rats in each group were randomly selected, and anaesthetized intraperitoneally with 3% pentobarbital sodium 0.15 mL/100 g body weight, the blood and organ samples were collected to analyze blood routine, serum electrolyte, serum biochemical indexes and organ coefficients. The heart, liver, kidney and lung of LDRH and BDRH group rats were fixed with 10% neutral formaldehyde (V/V), stained with Hematoxylin-eosin and examined by optical microscope to analyze the toxicity of DRR1. The delayed toxicity effect of DRR1 was performed on the 14^th day after drug withdrawal, by the same way as above groups. The behavior, food intake and body weight of rats were observed after administration and recovery period.Results After 30 days of administration with different concentrations of DRR1 living bacteria and its fragments, at the point of drug withdrawal 0 d and 14 d, no abnormal movement and respiration were observed, no abnormal phenomena such as slow reaction, pupil change, eyeball protrusion and skin color change were observed; no abnormal secretion was found in mouth, ear, nose and eye corner; no abnormallities were found in urination and defecation and stool morphology. Compared with TGY group, the food intake, body weight and organ coefficient of the DRR1 groups had no significant changes. The number of white blood cells (WBC) in LDRH, decreased significantly on the 30^th day of administration (P < 0.05), and returned to the TGY group level after 14 days of drug withdrawal. The number of red blood cells, hematocrit and hemoglobin had no significant changes, the number of lymphocytes, granulocytes, monocytes and their percentages had no significant changes, and the number of platelets had no significant changes. The serum alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein (TP), uric acid (UA), urea (UREA), blood glucose (Glu), phosphorus (P) and magnesium (Mg) were not significantly changed after drug withdrawal. The levels of aspartate aminotransferase (AST), creatine kinase (CK), creatine kinase MB (CK-MB) and lactate dehydrogenase (LDH) in LDRH group were significantly increased (P < 0.05). After 14 days of recovery, AST and CK decreased to the level of TGY, CK-MB was still higher than that of TGY group (P < 0.05); LDH in the recovery period was significantly higher than that 0 h after drug withdrawal (P < 0.05), no significant change compared with recovery period TGY group. There was no significant change in serum ion concentration (P > 0.05) on the 30th day of administration. On the 14th day after drug withdrawal, Cl^- and Ca²⁺ decreased, Na⁺ and pH increased (P < 0.05) in LDRH group. No significant pathological changes were found in heart, liver, lung and kidney of LDRH and BDRH group.Conclusion DRR1 live bacteria and its bacterial fragments are practically non-toxic on rats. This study provides experimental data for the application of DRR1.

Key words: Deinococcus radiodurans, Food intake, Bacterial debris, Chronic toxicity, SD rats

中图分类号:

Q95-33

许琴, 王玮, 董翔, 李建瑛, 是文辉, 刘晓禄, 刘爱中, 马娜, 宋来阳, 刘江伟. 耐辐射奇球菌R1对大鼠的慢性毒性实验[J]. 实验动物与比较医学, 2021, 41(4): 333-342.

XU Qin, WANG Wei, DONG Xiang, LI Jianying, SHI Wenhui, LIU Xiaolu, LIU Aizhong, MA Na, SONG Laiyang, LIU Jiangwei. Chronic Toxicity Study on Deinococcus Radiodurans R1 in Rats[J]. Laboratory Animal and Comparative Medicine, 2021, 41(4): 333-342.

图/表 7

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