吉非替尼按时辰给药的药理作用特点及其机制研究
发布时间：2018-09-03 14:08
【摘要】：目的初步探讨吉非替尼按时辰给药荷瘤小鼠的药理作用特点及其作用机制。 方法 1.建立肺癌小鼠模型并分组：利用C57BL/6小鼠建立lewis肺癌小鼠模型,将荷瘤小鼠随机分成13组(n=10),即A-F六个实验组、a-f六个空白组和对照组；A-F实验组分别在8：00、12：00、16：00、20：00、24：00、次日4：00以灌胃方式给药(100m·kg-1),空白组和对照组给予相同剂量的含有5%羧甲基纤维素钠的蒸馏水。 2.日常指标的测定：连续给药时间21天,每天观察并记录各组小鼠生存状态及出现肛周红肿的小鼠数量,每三天测定一次小鼠肿瘤体积,三周后在相应的时间进行眼眶取血并脱臼处死小鼠,剥离肿瘤并称量瘤重,计算抑瘤率。 3.血常规检测：各组每只荷瘤小鼠取50μl血液进行血常规检测。 4.肿瘤组织和皮肤组织的观察：对剥离的肿瘤同时进行病理学分析以及扫描电镜分析,同时对部分皮肤组织进行扫描电镜观察。 5.细胞因子的测定：应用ELISA技术(按照美国Biolegend公司试剂盒说明书的要求)测定不同组的小鼠血液中细胞因子IL-6. IL-2和TNF-a水平。 6.基因表达的测定：应用RT-PCR技术测定肿瘤组织中EGFR、MMP-9、ABCG2和P35基因表达。 结果 1.吉非替尼对小鼠的生存质量有一定的影响,A、B、F组的小鼠精神状态和身体状况比C、D、E组好,A组和F组荷瘤小鼠的肛周红肿率较其它实验组低。 2.吉非替尼可以明显抑制肿瘤的增长,在所有的实验组中,A组荷瘤小鼠的肿瘤体积增长最缓慢(P0.05),A组的抑瘤率最高,C组最低(44.12%vs14.15%,x2=36.00,P=0.000),F组次之；病理学分析和扫描电镜分析显示A组和F组肿瘤组织坏死最严重,该组的上皮细胞损伤程度较其它实验组轻。 3.血常规分析各组白细胞、红细胞、血红蛋白、中性粒细胞差异没有统计学意义(P0.05)；实验组小鼠血清中的IL-6、IL-2和TNF-a含量升高。A组和F组血清中的IL-6、IL-2和TNF-α水平最接近对照组的水平,C组和D组含量最高,差异显著(P0.05)。 4.时辰给药荷瘤小鼠对EGFR基因表达影响不同,A组EGFR表达最低,C、D组EGFR基因表达相对较高,差异有统计学意义(P0.05)。空白组间EGFR基因表达呈节律性变化,12:00左右时表达最高,20：00左右时EGFR表达最低。各实验组中MMP-9、ABCG2和P53基因表达有显著性差异,C组和D组表达最高,A组表达最低(P0.05)。结论 1.吉非替尼在荷瘤小鼠上的抗肿瘤活性和毒副作用存在时辰节律性,明期早期(8：00)和暗期晚期(4：00)的抗肿瘤活性较强,对小鼠的毒副作用相对较低。 2.吉非替尼药理学作用特点呈时辰节律性变化,其机制可能与荷瘤小鼠体内的EGFR呈节律性变化有关；应用吉非替尼时,应考虑时间因素的影响。
[Abstract]:Objective to investigate the pharmacological characteristics and mechanism of gifitinib in mice with tumor-bearing drugs. Method 1. To establish lung cancer mice model and group: the lewis lung cancer mice model was established by C57BL/6 mice. The tumor bearing mice were randomly divided into 13 groups, namely A-F six experimental groups, a-f six blank group and control group. A-F experimental group was given orally at 8: 00 at 12: 00 to 16: 00 at 20: 00: 24: 00, by gavage at 4:00 the next day (100m kg-1). The blank group and the control group were given the same dose of distilled water containing 5% carboxymethyl cellulose sodium. 2. Measurement of daily indexes: continuous administration for 21 days, observed and recorded the survival status and the number of mice with perianal redness and swelling in each group, and measured the tumor volume of mice every three days. Three weeks later, blood was taken from orbit and dislocated mice were killed at the corresponding time, tumor was stripped and weighed, and tumor inhibition rate was calculated. Blood routine examination: 50 渭 l blood samples were taken from each tumor-bearing mouse in each group. 4. 4. Observation of tumor tissue and skin tissue: pathological analysis and scanning electron microscope analysis of exfoliated tumor were carried out simultaneously, and some skin tissues were observed by scanning electron microscope at the same time. Determination of cytokines: determination of cytokine IL-6. in blood of different groups of mice by ELISA technique (as required by Biolegend kit specification) IL-2 and TNF-a levels. Detection of gene expression: RT-PCR technique was used to detect the expression of EGFR,MMP-9,ABCG2 and P35 genes in tumor tissues. Result 1. Gefitinib had a certain effect on the quality of life of mice. The mental state and physical condition of the BF group were lower than those of the other experimental groups. 2. The rate of perianal red swelling in group A and group F was lower than that in group A and F. Gefitinib could significantly inhibit tumor growth. In all the experimental groups, the tumor volume of group A was the slowest (P0.05), the inhibition rate of group C was the highest (44.12 vs 14.15x36.00 P0. 000) and that of group F was the lowest. Pathological analysis and scanning electron microscope analysis showed that tumor tissue necrosis was the most serious in group A and group F, and the degree of epithelial cell injury in group A was less than that in other experimental groups (3.3%). There was no significant difference in white blood cell, erythrocyte, hemoglobin, neutrophil in each group by blood routine analysis (P0.05). The levels of IL-6,IL-2 and TNF- 伪 in serum of group A and F were the highest in group C and group D (P0.05). Effect of time administration on the expression of EGFR gene in tumor-bearing mice the expression of EGFR gene in the lowest EGFR expression group (P 0.05) was significantly higher than that in the control group (P 0.05). There was a rhythmic change of EGFR gene expression in the blank group. The highest expression of EGFR gene was observed at about 12: 00 and the lowest at about 20: 00. There was significant difference in the expression of MMP-9,ABCG2 and p53 gene in each experimental group. The expression of MMP-9,ABCG2 and p53 gene was the lowest in group C and group D (P0.05). Conclusion 1. The antitumor activity and side effects of Gifitinib in tumor-bearing mice were rhythmic, and the antitumor activity in early stage (8:00) and late stage (4:00) was stronger, and the toxicity and side effect of gifitinib on mice was relatively low. 2. The pharmacological characteristics of gefitinib were time-rhythmic, and its mechanism might be related to the rhythmic change of EGFR in tumor-bearing mice, and the effect of time should be taken into account when gifetini was used.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
本文编号：2220197
[Abstract]:Objective to investigate the pharmacological characteristics and mechanism of gifitinib in mice with tumor-bearing drugs. Method 1. To establish lung cancer mice model and group: the lewis lung cancer mice model was established by C57BL/6 mice. The tumor bearing mice were randomly divided into 13 groups, namely A-F six experimental groups, a-f six blank group and control group. A-F experimental group was given orally at 8: 00 at 12: 00 to 16: 00 at 20: 00: 24: 00, by gavage at 4:00 the next day (100m kg-1). The blank group and the control group were given the same dose of distilled water containing 5% carboxymethyl cellulose sodium. 2. Measurement of daily indexes: continuous administration for 21 days, observed and recorded the survival status and the number of mice with perianal redness and swelling in each group, and measured the tumor volume of mice every three days. Three weeks later, blood was taken from orbit and dislocated mice were killed at the corresponding time, tumor was stripped and weighed, and tumor inhibition rate was calculated. Blood routine examination: 50 渭 l blood samples were taken from each tumor-bearing mouse in each group. 4. 4. Observation of tumor tissue and skin tissue: pathological analysis and scanning electron microscope analysis of exfoliated tumor were carried out simultaneously, and some skin tissues were observed by scanning electron microscope at the same time. Determination of cytokines: determination of cytokine IL-6. in blood of different groups of mice by ELISA technique (as required by Biolegend kit specification) IL-2 and TNF-a levels. Detection of gene expression: RT-PCR technique was used to detect the expression of EGFR,MMP-9,ABCG2 and P35 genes in tumor tissues. Result 1. Gefitinib had a certain effect on the quality of life of mice. The mental state and physical condition of the BF group were lower than those of the other experimental groups. 2. The rate of perianal red swelling in group A and group F was lower than that in group A and F. Gefitinib could significantly inhibit tumor growth. In all the experimental groups, the tumor volume of group A was the slowest (P0.05), the inhibition rate of group C was the highest (44.12 vs 14.15x36.00 P0. 000) and that of group F was the lowest. Pathological analysis and scanning electron microscope analysis showed that tumor tissue necrosis was the most serious in group A and group F, and the degree of epithelial cell injury in group A was less than that in other experimental groups (3.3%). There was no significant difference in white blood cell, erythrocyte, hemoglobin, neutrophil in each group by blood routine analysis (P0.05). The levels of IL-6,IL-2 and TNF- 伪 in serum of group A and F were the highest in group C and group D (P0.05). Effect of time administration on the expression of EGFR gene in tumor-bearing mice the expression of EGFR gene in the lowest EGFR expression group (P 0.05) was significantly higher than that in the control group (P 0.05). There was a rhythmic change of EGFR gene expression in the blank group. The highest expression of EGFR gene was observed at about 12: 00 and the lowest at about 20: 00. There was significant difference in the expression of MMP-9,ABCG2 and p53 gene in each experimental group. The expression of MMP-9,ABCG2 and p53 gene was the lowest in group C and group D (P0.05). Conclusion 1. The antitumor activity and side effects of Gifitinib in tumor-bearing mice were rhythmic, and the antitumor activity in early stage (8:00) and late stage (4:00) was stronger, and the toxicity and side effect of gifitinib on mice was relatively low. 2. The pharmacological characteristics of gefitinib were time-rhythmic, and its mechanism might be related to the rhythmic change of EGFR in tumor-bearing mice, and the effect of time should be taken into account when gifetini was used.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
【参考文献】
相关期刊论文 前10条
1 杜志明;吴志诚;江涛;贺方兴;江柏青;;VEGF-C和ABCG2在非小细胞肺癌中的表达及意义[J];赣南医学院学报;2010年02期
2 张靖;马虎;韩静;李雪涛;陈正堂;;ABCG2在非小细胞肺癌厄洛替尼耐药中的作用[J];第三军医大学学报;2014年10期
3 张宇飞;郭丽;赵峰;姜晓玲;穆德广;徐兴祥;戚好文;;ABCG2在肺癌中的表达及意义[J];解放军医学杂志;2007年09期
4 亓梅;赵慧;王翠萍;;P53与肺癌诊断治疗研究进展[J];临床肺科杂志;2007年11期
5 韩雪;戴广海;;胃癌中EGFR相关研究及意义[J];临床肿瘤学杂志;2010年02期
6 乔建兵;陈文萍;;比较吉非替尼与化疗一线治疗晚期非小细胞肺癌的Meta分析[J];临床肿瘤学杂志;2013年05期
7 周守兵;蒋华;李文娟;房新建;;培美曲塞时辰给药治疗肺腺癌裸鼠移植瘤的疗效[J];实用医学杂志;2012年14期
8 柯立;徐美青;魏大中;马冬春;;非小细胞肺癌中S100A2和p53蛋白的表达[J];安徽医科大学学报;2010年04期
9 张晓梅;钟建明;汤敏中;张晓光;廖建;郑裕明;邓洪;曾毅;;EBV 4型IgA/VCA IgA/EA IgG/EA IgG/ZEBRA抗体在鼻咽癌普查和早期诊断中的应用[J];中华实验和临床病毒学杂志;2006年03期
10 冯大鹏;肖建如;;基质金属蛋白酶及其抑制剂与肿瘤的相关性[J];肿瘤;2009年09期
，本文编号：2220197
本文链接：https://www.wllwen.com/yixuelunwen/yiyaoxuelunwen/2220197.html
最近更新
教材专著
