巴豆醛对雄性大鼠的毒性研究

发布时间：2018-06-26 11:25

本文选题：巴豆醛 + 雄性大鼠　；参考：《济南大学》2016年硕士论文

【摘要】：巴豆醛(Crotonaldehyde),其化学结构式为CH3CH=CHCHO,其有顺式和反式两种双键异构体。通常为无色或略带黄色的液体,可以燃烧,具有催泪性,暴露于空气中或遇到光照时逐渐变为淡黄色液体,随即被氧化为巴豆酸。巴豆醛可经口、鼻及皮肤等途径侵入机体,具有明显的刺激作用和催泪作用。然而,国内外对巴豆醛的动物毒理学实验研究资料极少,因此本实验以雄性大鼠为对象进行毒研究,旨在全面探讨巴豆醛的毒理学损伤特性。【研究目的】初步探讨巴豆醛对雄性大鼠的毒性损伤作用,为近一步研究巴豆醛对作业人群的生物监测和健康监护方案提供重要依据和线索,并弥补国内、外相关资料的缺失。【研究方法】1.巴豆醛对大鼠的急性经口毒性研究选择SPF级健康Wistar大鼠40只,体重在180~220克之间按照雌雄各分为4组,每组5只,动物禁食12小时后灌胃给药,根据预试验结果设计染毒剂量为215、100、46.4、21.5mg/kg。动物染毒后连续观察14天,其间观察和记录动物中毒症状及死亡情况,最后根据每组动物数、组距和每组动物死亡数,即可从霍恩氏表中查得半数致死量(LD50)及其95%可信限。2.巴豆醛对大鼠的蓄积毒性研究本次实验选取40只健康Wistar大鼠,随机分为2组,每组20只,雌雄各半,采用固定剂量法染毒,实验组大鼠每天经口染毒1/5LD50剂量的巴豆醛,连续灌胃染毒28d,对照组大鼠给予同剂量的溶剂。实验期内大鼠累计死亡数达50%,终止实验。按照公式计算蓄积系数(K),以评价化合物对机体蓄积性毒性的大小。3.巴豆醛对雄性大鼠的亚慢性毒性研究选取SPF级健康Wistar雄性大鼠40只,体重在190~200克之间,待大鼠适应性饲养1周后,随机分为4组,每组10只,分为高、中、低3个剂量组(染毒剂量分别为8.44mg/kg、4.22mg/kg、2.11mg/kg)和1个对照组,对照组给予蒸馏水处理。经口灌胃,每天1次,连续染毒28d。最后一次染毒结束24h后,采用颈椎脱臼法处死大鼠,迅速摘取大鼠主要脏器,称重并计算脏器系数。摘眼球取血法采取大鼠外周血,全自动血生化分析仪测血清生化指标;酶联免疫吸附法(elisa法)测定氧化损伤指标;常规he染色观察肝组织病理学改变;流式细胞术(fcm)进行淋巴细胞亚群分析;酶联免疫吸附法(elisa)检测大鼠外周血及组织中细胞因子表达水平。【研究结果】1.巴豆醛对大鼠的急性经口毒性及蓄积毒性研究查霍恩氏表可得,巴豆醛对雌雄wistar大鼠的急性经口ld50如下:雌性大鼠为147mg/kg,95%可信限为95.1-227mg/kg;雄性大鼠为42.2mg/kg,95%可信限为25.3-70.2mg/kg。按照化学品毒理学分级标准,巴豆醛为高毒化学品。采用固定计量法,根据公式求得蓄积系数k5,认为巴豆醛有轻度蓄积毒性。2.巴豆醛对雄性大鼠的亚慢性毒性研究与对照组相比,高剂量组大鼠体重增重减少,肝脏脏器系数明显增高,且随着染毒剂量的增加,肝脏系数有增高的趋势;血清中谷丙转氨酶(alt)和谷草转氨酶(ast)水平明显增加,高剂量组和中剂量组的mda含量升高,sod含量降低,高剂量组的gsh-px含量降低,差异均有统计学意义(p0.05);与对照组相比,心脏系数、脾脏系数、肾脏系数及血清中总胆红素(tbil)、尿素(bun)、尿酸(ua)、肌酐(cre)、葡萄糖(glu)、甘油三酯(tg)、总胆固醇(chol)水平无明显变化,差异无统计学意义(p0.05);病理组织切片显示大鼠肝组织有炎性损伤;与对照组比较,各剂量组大鼠外周血中cd3+cd8+(%)、cd3+cd4+(%)及cd3(%)均无明显改变,差异无统计学意义(p0.05);与对照组比较,各组大鼠血清中il-4、il-6和tnf-α组间含量差异有统计学意义(p0.05),且随着染毒剂量的增加,各组il-4、il-6和tnf-α的含量呈升高的趋势,且高剂量组变化最为明显;与对照组比较,各组大鼠血清中il-1b及ifn-γ组间变化不明显,差异无统计学意义(p0.05);与对照组比较,各组大鼠肝脏组织中il-4、il-6、ifn-γ和tnf-α组间含量差异有统计学意义(p0.05),且随着染毒剂量的增加,各组l-4、il-6、ifn-γ和tnf-α的含量呈升高的趋势;与对照组比较,各组大鼠肝脏组织中il-1b组间变化不明显,差异无统计学意义(p0.05);与对照组比较,各组大鼠肺脏组织中il-1b组间含量差异有统计学意义(p0.05),且随着染毒剂量增加,il-1b含量也增加,高剂量组变化最为显著;与对照组比较,各组大鼠肺脏组织中IL-4、IL-6、IFN-γ和TNF-α组间变化不明显,差异无统计学意义(P0.05);与对照组比较,各组大鼠肾脏组织中IL-1b、IL-4、IL-6、IFN-γ和TNF-α的表达水平组间变化不明显,差异均无统计学意义(P0.05)。【研究结论】1.巴豆醛对雌雄Wistar大鼠的急性经口LD50如下:雌性大鼠为147mg/kg,95%可信限为95.1-227mg/kg;雄性大鼠为42.2mg/kg,95%可信限为25.3-70.2mg/kg。按照化学品毒理学分级标准,巴豆醛属于高毒化学品,且对雄性大鼠较为敏感。根据蓄积毒性实验结果(K5),可以认为巴豆醛对大鼠有轻度蓄积毒性,为下一步进行亚急性毒性研究的染毒剂量选择提供了参考依据。2.巴豆醛经口染毒导致雄性大鼠体重增重下降,肝脏的脏器系数明显增加。3.巴豆醛对雄性大鼠肝脏组织造成病理损伤,且损伤程度随染毒剂量的增加而增高。4.巴豆醛导致雄性大鼠血液中生化指标ALT、AST的升高,反应了肝脏受损。氧化应激指标MDA含量升高,SOD含量降低,说明机体受到自由基攻击。5.巴豆醛染毒导致肝组织匀浆中IL-4、IL-6、IFN-γ和TNF-α水平升高,说明巴豆醛对肝脏组织有炎性损伤。
[Abstract]:Crotonaldehyde, its chemical structural formula is CH3CH=CHCHO, which has two kinds of CIS and trans isomers. Usually colorless or slightly yellow liquid, it can be burned, teary, exposed to air or when exposed to light, gradually changed into yellowish liquid, which is oxidized to bean acid. The way to invade the body has obvious stimulating effect and leardrop effect. However, there are few experimental data on animal toxicology of croton aldehyde at home and abroad. Therefore, this experiment took the male rats as the object to study the toxicological properties of the soybean aldehyde in an all-round way. The effect of toxic damage can provide an important basis and clue to study the biological monitoring and health monitoring program of the operating crowd, and make up for the lack of related data in China. [method] 1. the study of acute oral toxicity of 1. soybean aldehyde to rats, 40 rats of SPF grade healthy Wistar were selected and the weight was between 180~220 grams. The female and male were divided into 4 groups, each group was 5. After fasting for 12 hours, the animals were given the medicine for 12 hours. According to the results of the pre test, the poisoned dose was observed for 14 days. The symptoms and death conditions of the animals were observed and recorded. Finally, the number of animals in each group, the distance from the group and the number of death in each group could be obtained from Hun's family. 40 healthy Wistar rats were randomly divided into 2 groups, 20 rats in each group were randomly divided into 2 groups, 20 in each group and half of male and male in each group. The experimental group was infected with fixed dose method. The experimental group was exposed to 1/ 5LD50 dose of Wistar aldehyde in the experimental group every day. The rats were continuously exposed to 28d and the control group. The rats were given the same dose of solvent. The cumulative death number of rats in the experimental period was 50% and the experiment was terminated. The cumulative coefficient of accumulation (K) was calculated according to the formula to evaluate the size of the cumulative toxicity of the compound to the body.3.. The subchronic toxicity study of the male rats was selected by the SPF grade healthy Wistar male rats. The weight of the male rats was in the 190~200 gram, and the rat was suitable for the rat. After 1 weeks of sexual rearing, they were randomly divided into 4 groups, 10 in each group, which were divided into high, middle and low 3 doses group (8.44mg/kg, 4.22mg/kg, 2.11mg/kg) and 1 control groups. The control group was treated with distilled water. After the oral administration of the stomach, the rats were infected with 28d. for the last time 24h, and the rats were killed by dislocated cervical vertebra and exiexed quickly. The main organs of the rat were weighed and the organ coefficients were weighed. The peripheral blood of rats was taken by taking the eyeball extraction method, the biochemical indexes of the serum were measured by the automatic blood biochemical analyzer; the oxidative damage index was measured by the enzyme linked immunosorbent assay (ELISA method); the pathological changes of liver tissues were observed by the routine HE staining; the lymphocyte subsets were analyzed by flow cytometry (FCM); enzyme linked immunization The expression level of cytokine in the peripheral blood and tissue of rats was detected by ELISA. [results] 1. the acute oral toxicity and accumulation toxicity of croton aldehyde to rats was obtained. The acute oral LD50 of the female and male Wistar rats was as follows: the female rats were 147mg/kg, and the male rats were 95.1-227mg/kg; the male rats were 95.1-227mg/kg. For 42.2mg/kg, the 95% confidence limit is 25.3-70.2mg/kg. according to the classification standard of Chemical Toxicology and the soybean aldehyde is a high toxic chemical. The fixed measurement method is used to obtain the accumulation coefficient K5 according to the formula. It is believed that the subchronic toxicity of the soybean aldehyde with mild cumulative toxic.2. to the male rats is compared with the control group, the weight gain of the high dose group is increased. The coefficient of liver organs increased significantly, and the liver coefficient increased with the increase of dose. The level of ALT and AST in the serum increased significantly, the MDA content in the high dose group and the middle dose group increased, the content of SOD decreased, and the GSH-Px content in the high dose group decreased, the difference was statistically significant (p0.0 5): compared with the control group, the cardiac coefficient, spleen coefficient, kidney coefficient and serum total bilirubin (TBIL), urea (BUN), uric acid (UA), creatinine (CRE), glucose (Glu), triglyceride (TG), total cholesterol (Chol) level had no significant change, and the difference was not statistically significant (P0.05); pathological tissue section showed inflammatory injury in rat liver tissue; and control group. There was no significant change in cd3+cd8+ (%), cd3+cd4+ (%) and CD3 (%) in the peripheral blood of rats in each dose group, and there was no significant difference (P0.05). Compared with the control group, there was a significant difference in the content of IL-4, IL-6 and tnf- alpha in the serum of each group (P0.05), and the content of IL-4, IL-6 and tnf- alpha in each group increased with the increase of the dose. There was no significant change between IL-1B and ifn- gamma groups in the serum of rats in the control group, and there was no significant difference (P0.05). Compared with the control group, the differences in the content of IL-4, IL-6, ifn- gamma and tnf- alpha in the liver tissues of the rats were statistically significant (P0.05), and with the increase of dosage, each group had a significant difference (P0.05). The content of L-4, IL-6, ifn- gamma and tnf- alpha in the group of rats was increased. Compared with the control group, there was no significant difference between the IL-1B groups in the liver tissues of the rats (P0.05). Compared with the control group, the difference in the content of the IL-1B groups in the lung tissues of the rats in each group had a statistical significance (P0.05), and as the dose increased, the IL-1B content was also increased. In the control group, the changes of IL-4, IL-6, IFN- gamma and TNF- a were not significant (P0.05). Compared with the control group, there was no significant difference in the expression of IL-1b, IL-4, IL-6, IFN- gamma and TNF- alpha in the kidney tissues of the rats. The differences were not statistically significant, and the difference was not statistically significant. Significance (P0.05). [Conclusion] [Conclusion] 1. the acute oral LD50 of the female and male Wistar rats is as follows: the female rats are 147mg/kg, the 95% confidence limit is 95.1-227mg/kg, the male rats are 42.2mg/kg, the 95% confidence limit is 25.3-70.2mg/kg. according to the chemical toxicology grading standard, and the soybean aldehyde belongs to the high toxic chemical, and it is more sensitive to the male rats. According to the results of the cumulative toxicity test (K5), it can be considered that the croton aldehyde has a mild cumulative toxicity to rats, which provides a reference basis for the next step of subacute toxicity study, which provides a reference basis for the weight gain of the male rats induced by the oral administration of.2., and the organ coefficient of the liver is significantly increased by the.3. of the male rat liver. The pathological damage was caused by the histopathology, and the damage degree increased with the increase of the dose of.4., which resulted in the increase of ALT and AST in the blood of the male rats and the liver damage. The oxidative stress index MDA content increased and the SOD content decreased, indicating that the body was exposed to the free radical attack of.5., which resulted in the IL-4, IL-6, IFN- in the liver homogenate. Elevated levels of gamma and TNF- alpha indicate that croton aldehyde has inflammatory injury to liver tissue.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R114

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