摘要
【目的】明确呼宁散的毒理作用,评价其安全性,为其用于临床治疗鸡呼吸道疾病综合征提供科学依据。【方法】急性毒性预试验中,将24只小鼠随机均分为对照组和呼宁散低、中、高剂量组,呼宁散低、中、高剂量组小鼠分别灌胃2.5、5、10 g/kg BW呼宁散1次,0.2 mL/10 g BW,对照组灌胃等体积灭菌蒸馏水,连续观察7 d,记录小鼠中毒和死亡情况。最大给药量试验中,将20只小鼠随机均分为对照组和试验组,试验组小鼠在24 h内分4次灌胃呼宁散10 g/kg BW,0.2 mL/10 g BW,对照组灌胃等体积灭菌蒸馏水,于试验第15天记录小鼠体重并剖检,观察小鼠主要脏器病理变化。亚急性毒性试验中,将40只小鼠随机均分为对照组和呼宁散低、中、高剂量组(2.5、5和10 g/kg BW),连续给药28 d,于末次给药12 h后对小鼠称重、采血,剖检并采集心脏、肝脏、脾脏、肺脏、肾脏,计算小鼠体重变化、脏器系数,检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-4(IL-4)和IL-6含量,并对心脏、肝脏、脾脏、肺脏、肾脏进行组织病理学检查。在骨髓微核试验中,将50只小鼠随机均分为对照组、阳性对照组(环磷酰胺40 mg/kg BW)和呼宁散低、中、高剂量组(2.5、5和10 g/kg BW),各组小鼠灌胃给药2次(间隔24 h),末次给药6 h后处死小鼠并制作骨髓涂片,计算微核率及嗜多染红细胞(PCE)/成熟红细胞(NCE)值。在精子畸形试验中,将25只雄性小鼠随机均分为对照组、阳性对照组(环磷酰胺40 mg/kg)和呼宁散低、中、高剂量组(2.5、5和10 g/kg BW),每天灌胃给药1次,连续给药5 d,于首次灌胃后35 d处死小鼠并制作精子涂片,计算精子畸形率。【结果】急性毒性试验中,各组小鼠全部存活,均未见中毒症状,无法测出呼宁散的半数致死量(LD 50),测得小鼠对呼宁散的最大耐受量>40 g/kg BW,说明呼宁散对小鼠无急性毒性。亚急性毒性试验中,与对照组雄性小鼠相比,给药第7天,呼宁散低剂量组小鼠体重显著增加(P<0.05);给药第28天,呼宁散中剂量组小鼠体重极显著增加(P<0.01);呼宁散对各组小鼠主要脏器系数、细胞因子含量均无显著影响(P>0.05),剖检和病理学检查未见明显异常变化,说明呼宁散无亚急性毒性。骨髓细胞微核试验和精子畸形试验中,各剂量呼宁散对小鼠骨髓微核率及精子畸形率无显著影响(P>0.05),且PCE/NCE值均在正常范围内,说明呼宁散无遗传毒性。【结论】10 g/kg BW及以下剂量呼宁散无毒性作用,安全性好。
【Objective】The aim of this study was to determine the toxicity and safety of Huning powder and provide scientific basis for its clinical application in the treatment of chicken respiratory disease syndrome.【Method】In the acute toxicity test,24 mice were randomly divided into control group and Huning powder low,medium and high doses of groups.Mice in Huning powder low,medium and high dose groups were given 2.5,5 and 10 g/kg BW Huning powder once with 0.2 mL/10 g BW,respectively,while mice in control group were given equal volume of sterilized distilled water.The toxicity and death of the mice were observed for 7 days.In the maximum dose test,20 mice were randomly divided into control group and test group.Mice in test group were given 10 g/kg BW Huning powder with 0.2 mL/10 g BW for 4 times within 24 h,mice in control group were given equal volume of sterilized distilled water.On the 15th day of the trial,the body weight was recorded and the pathological changes of main organs were observed.In the subacute toxicity test,40 mice were randomly divided into control group and Huning powder low,medium and high doses of groups(2.5,5 and 10 g/kg BW),all mice were administered continuously for 28 days,at 12 h after the last administer,mice were weighed,blood was collected,dissected and the heart,liver,spleen,lung and kidney were collected,the change of body weight and organ coefficient of mice were calculated,the contents of tumor necrosis factor-α(TNF-α),interleukin-4(IL-4)and IL-6 in serum of mice were determined and histopathological examination was performed for the heart,liver,spleen,lung and kidney.In the bone marrow micronucleus test,50 mice were randomly divided into control group,positive control group(cyclophosphamide 40 mg/kg BW)and Huning powder low,medium and high doses of groups(2.5,5 and 10 g/kg BW),mice in each group were administered 2 times(interval 24 h),at 6 h after the last administer,mice were sacrificed and bone marrow smears were made,the rate of micronucleus and PCE/NCE value were calculated.In the sperm abnormality test,25 male mice were randomly divided into control group,positive control group(cyclophosphamide 40 mg/kg BW)and Huning powder low,medium and high doses of groups(2.5,5 and 10 g/kg BW),administered once a day for 5 days,the mice were sacrificed on day 35 after first administer and sperm smears were made,the rate of sperm deformity was calculated.【Result】In the acute toxicity test,all mice in each group were survived without any toxic symptoms,and the LD 50 of Huning powder was not obtained,the maximum tolerance of Huning powder in mice was 40 g/kg BW,indicating that the Huning powder was safe and had no acute toxicity.In the subacute toxicity test,compared with male mice in control group,the weight of mice in Huning powder low dose group were significantly increased on the 7th day of administration(P<0.05),the weight of mice in Huning powder medium dose group were extremely significantly increased on the 28th day of administration(P<0.01),and Huning powder had no significant effect on the main organ coefficients and cytokine contents of mice in each group(P>0.05),necropsy and pathological examination showed no obvious abnormal changes,indicating that Huning powder had no subacute toxicity.In the bone marrow micronucleus test and sperm abnormality test,Huning powder did not affect the bone marrow micronucleus rate and sperm deformity rate of mice(P>0.05),and the PCE/NCE values were all within the normal range,indicating that Huning powder had no genetic toxicity.【Conclusion】Huning powder at the dose 10 g/kg BW and below was safe and nontoxic.
作者
王巍
桑锐
葛冰洁
闫可心
刘馨蔓
于明弘
张雪梅
WANG Wei;SANG Rui;GE Bingjie;YAN Kexin;LIU Xinman;YU Minghong;ZHANG Xuemei(Agriculture College of Yanbian University,Yanji 133002,China)
出处
《中国畜牧兽医》
CAS
CSCD
北大核心
2023年第12期5222-5231,共10页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(32060817)
吉林省科技发展计划重点项目(20210202034NC)。
关键词
呼宁散
小鼠
急性毒性
亚急性毒性
遗传毒性
Huning powder
mice
acute toxity
subcaute toxicity
genetic toxicity