母鼠维生素D缺乏及干预对仔鼠肺发育及VEGF、SP-C表达影响的研究
发布时间：2018-07-26 07:58
【摘要】：目的:通过建立Vit D缺乏及Vit D干预母鼠模型,观察母鼠孕期、哺乳期Vit D缺乏及适量1,25二羟维生素D3[1,25-(OH)2D3]干预对其仔鼠肺发育及肺血管内皮生长因子(VEGF)、肺组织表面活性蛋白C(SP-C)的变化,从而探讨母鼠Vit D缺乏及适量干预对仔鼠肺发育的影响及可能机制。方法:选取健康成年雌性清洁级SD大鼠,完全随机分为正常对照组(对照组)、Vit D缺乏组(缺乏组)、1,25-(OH)2D3干预组(干预组),每组6只。对照组及干预组母鼠予以正常饲料、正常光照喂养,缺乏组母鼠予不含Vit D饲料、避光喂养,试喂2周后,用ELISA法测定三组母鼠血清25-(OH)D值,再与正常成年雄性SD大鼠合笼交配。于受孕第7天开始,干预组母鼠隔天一次给予1,25-(OH)2D3(10ug/kg)灌胃,对照组及缺乏组母鼠予1ml生理盐水代替灌胃,一直到分娩后第21天,再次测三组母鼠血清25-(OH)D值,并检测各组仔鼠生后第1天及第21天血清25-(OH)D值,再取生后第1、7、14、21天仔鼠肺组织,观察以下指标:1.仔鼠体重及肺湿重;2.光镜下观察肺形态结构;3.q-PCR法检测肺组织VEGF mRNA水平的表达;4.免疫组织化学方法分别检测肺组织VEGF、SP-C蛋白表达。结果:1.造模结果:通过不同方式喂养母鼠,试喂2周后三组母鼠之间血清25-(OH)D值比较,差异无统计学意义(P0.05);分娩后第21天,缺乏组母鼠血清25-(OH)D值明显低于对照组及干预组母鼠,有显著差异(P0.01),干预组母鼠血清25-(OH)D值高于对照组母鼠,差异有统计学意义(P0.05);新生第1天仔鼠三组之间血清25-(OH)D值比较,差异无统计学意义(P0.05);新生第21天,缺乏组仔鼠血清25-(OH)D值低于对照组及干预组仔鼠,差异有统计学意义(P0.05),干预组仔鼠血清25-(OH)D值略高于对照组仔鼠,但差异无统计学意义(P0.05)。2.三组仔鼠体重及肺湿重结果:随着日龄的增加,缺乏组仔鼠体重、肺湿重均低于同日龄对照组仔鼠,增长速度缓慢,缺乏组第14、21天仔鼠体重较对照组仔鼠轻,差异有统计学意义(P0.05),第7、14、21天仔鼠肺湿重较对照组仔鼠轻,有显著差异(P0.01);干预组仔鼠体重、肺湿重均高于同日龄对照组仔鼠,增长速度稍快,其体重较对照组仔鼠比均有差异(P0.05,0.01),第14、21天仔鼠的肺湿重较对照组仔鼠比有差异(P0.05)。3.三组仔鼠HE染色光镜观察结果:缺乏组仔鼠较同日龄对照组仔鼠比肺组织肺泡数少,肺泡腔小,形态较不规则,实性团状结构区域增加,肺泡间隔较宽;干预组仔鼠较同日龄对照组仔鼠比肺组织肺泡数稍增加,肺泡腔大小均匀,实性团状结构区域减少,肺泡间隔较薄。4.q-PCR法检测三组仔鼠肺组织VEGF mRNA表达结果:随着日龄的增加,对照组及干预组仔鼠肺组织VEGF mRNA的表达量逐渐增多,缺乏组仔鼠生后第1天表达量高于对照组及干预组仔鼠,差异有统计学意义(P0.05,0.01),生后第7天较前下降,表达量明显低于对照组及干预组仔鼠,有显著差异(P0.01),之后表达量逐渐增加,但增长幅度较对照组及干预组仔鼠缓慢,增长曲线与正常组及干预组仔鼠有交叉现象;干预组仔鼠增长幅度较对照组仔鼠稍快,表达量均高于对照组仔鼠,差异有统计学意义(P0.05,0.01)。5.免疫组织化学方法检测三组仔鼠肺组织VEGF蛋白表达结果:随着日龄的增加,对照组及干预组仔鼠肺组织VEGF蛋白的表达量逐渐增多,缺乏组仔鼠生后第1天表达量较对照组及干预组仔鼠略高,生后第7天较前下降,低于对照组及干预组仔鼠,差异有统计学意义(P0.05,0.01),之后表达量逐渐增加,增长幅度较对照组及干预组仔鼠缓慢;干预组仔鼠增长幅度较对照组仔鼠稍快,表达量均高于对照组仔鼠,第14、21天仔鼠之间差异有统计学意义(P0.05)。6.免疫组织化学方法检测三组仔鼠肺组织SP-C表达结果:三组仔鼠随着日龄的增加,肺组织SP-C的表达量逐渐增加,第21天最高;缺乏组仔鼠肺组织SP-C表达量均低于同日龄对照组及干预组仔鼠,差异有统计学意义(P0.05,0.01);干预组仔鼠肺组织SP-C表达量较同日龄对照组仔鼠高,第14、21天仔鼠之间差异有统计学意义(P0.05)。结论:1.母鼠孕期、哺乳期Vit D缺乏会抑制其仔鼠生后的肺发育过程,若给予适量1,25-(OH)2D3干预,对仔鼠的肺发育过程有一定帮助。2.母鼠孕期、哺乳期Vit D缺乏及适量补充1,25-(OH)2D3可能通过影响仔鼠肺组织VEGF、SP-C的表达,从而干扰其肺发育。
[Abstract]:Objective: To observe the changes of lung development and pulmonary vascular endothelial growth factor (VEGF) and lung tissue surface active protein C (D) in the mother rat by establishing Vit D deficiency and Vit D intervention in the female rat model. The effect and possible mechanism of rat lung development. Methods: select healthy adult female clean SD rats, totally randomly divided into normal control group (control group), Vit D deficiency group (lack of group), 1,25- (OH) 2D3 intervention group (intervention group), 6 rats in each group. The control group and the intervention group were given normal diet, normal light was fed, and the lack of female rats were given no Vit D feeding. After feeding for 2 weeks, the serum 25- (OH) D value of the female rats was measured by ELISA method and then copated with the normal adult male SD rats. On the seventh day of pregnancy, the female mice in the intervention group were given 1,25- (OH) 2D3 (10ug/kg) every other day. The control group and the lack of the female rats were given 1ml saline instead of the stomach, until the twenty-first day after childbirth, again, again after the childbirth, again, again after the labor after the labor, again, again after the childbirth, again, again after the childbirth, again, after the labor, again, again after the labor after the twenty-first days, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again after delivery, again after delivery, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again after delivery, again after delivery, again after the birth, again after the birth, again, after the twenty-first day after childbirth, again The serum 25- (OH) D values were measured in three groups of female rats, and the serum 25- (OH) D values were measured at first days and 21 days after birth. The following indexes were observed: 1. mice weight and lung wet weight; 2. light microscopy was used to observe lung morphological structure; 3.q-PCR method was used to detect the expression of VEGF mRNA in lung tissue; 4. immunohistochemical methods were used. Do not detect the expression of VEGF and SP-C protein in lung tissue. Results: 1. the results of the model: feeding mice in different ways, after feeding 2 weeks, the serum 25- (OH) D value of the three groups of mice was not statistically significant (P0.05); the serum 25- (OH) D value of the lack of female mice was significantly lower than that of the control group and the intervention group, and there were significant differences (P0.01), intervention (P0.01). The serum 25- (OH) D value of the female rats was higher than that of the control group, the difference was statistically significant (P0.05); there was no significant difference in serum 25- (OH) D value between the first days of the newborn offspring of the newborn rats (P0.05), and the serum 25- (OH) D of the newborn rats was lower than that of the group and the intervention group (P0.05) in the twenty-first day of new birth (P0.05). The value of 25- (OH) D was slightly higher than that of the control group, but the difference was not statistically significant (P0.05) in.2. three groups of offspring rats weight and lung wet weight results: with the increase of age, the weight of the rats and the wet weight of lung were lower than those of the same age group, and the growth rate was slow, and the weight of 14,21 day offspring was less than that of the control group (P 0.05) the wet weight of lung in the 7,14,21 day rats was lighter than that of the control group (P0.01), and the weight of the rats in the intervention group was higher than that of the same age control group, and the growth rate was slightly faster than that of the control group (P0.05,0.01). The wet weight of the lungs in day 14,21 mice was significantly higher than that of the control group (P0.05).3. three groups. The results of HE staining light microscopy showed that the number of alveoli in the lung tissue was less than that of the same age group, the alveolus was small, the shape of the pulmonary alveolus increased and the interval of alveoli was wider, and the number of alveoli in the intervention group was slightly higher than that of the lung tissue in the control group, and the size of the alveolus was uniform and the solid structure area was reduced. The expression of VEGF mRNA in lung tissue of three groups of young rats was detected by.4.q-PCR method. The expression of VEGF mRNA in lung tissue of the control group and the intervention group increased gradually with the increase of day age. The first days after birth, the expression amount was higher than that of the control group and the intervention group. The difference was statistically significant (P0.05,0.01) and seventh days after birth. The rate of expression was significantly lower than that of the control group and the intervention group (P0.01), and the expression amount increased gradually, but the increase was slower than that of the control group and the intervention group. The growth curve was crossed with the normal group and the intervention group. The growth rate of the intervention group was faster than the control group. The expression amount was higher than that of the control group. The difference was statistically significant (P0.05,0.01).5. immunohistochemical method to detect the expression of VEGF protein in the lung tissue of three groups of offspring: with the increase of age, the expression of VEGF protein in the lung tissue of the control group and the intervention group increased gradually, and the expression of the first days after birth was slightly higher than that of the control group and the intervention group, seventh after birth, and seventh after birth. The difference between the control group and the intervention group was significantly lower than that of the control group and the intervention group (P0.05,0.01), and the expression amount increased gradually, and the growth amplitude was slower than that of the control group and the intervention group. The increase of the offspring rats in the intervention group was a little faster than the control group, and the expression amount was higher than that in the control group, and the difference was statistically significant (P). 0.05).6. immunohistochemical method was used to detect the expression of SP-C in lung tissue of three groups of offspring: the expression of SP-C in lung tissue increased gradually with the increase of day age in the three groups, and the expression of SP-C in lung tissue was lower than that of the same age group and the intervention group. The difference was statistically significant (P0.05,0.01). The expression of SP-C in lung tissue was higher than that of the same age control group, and there was a significant difference between the young rats at 14,21 day (P0.05). Conclusion: 1. mother rats during pregnancy, the lack of Vit D during lactation could inhibit the development of lung development of their offspring. If appropriate 1,25- (OH) 2D3 intervention was given, the lung development process of the offspring was certain to help.2. female rats and mammalian Vit D. Lack and proper supplement of 1,25- (OH) 2D3 may interfere with lung development by affecting the expression of VEGF and SP-C in the lung tissue of offspring rats.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.2
本文编号：2145373
[Abstract]:Objective: To observe the changes of lung development and pulmonary vascular endothelial growth factor (VEGF) and lung tissue surface active protein C (D) in the mother rat by establishing Vit D deficiency and Vit D intervention in the female rat model. The effect and possible mechanism of rat lung development. Methods: select healthy adult female clean SD rats, totally randomly divided into normal control group (control group), Vit D deficiency group (lack of group), 1,25- (OH) 2D3 intervention group (intervention group), 6 rats in each group. The control group and the intervention group were given normal diet, normal light was fed, and the lack of female rats were given no Vit D feeding. After feeding for 2 weeks, the serum 25- (OH) D value of the female rats was measured by ELISA method and then copated with the normal adult male SD rats. On the seventh day of pregnancy, the female mice in the intervention group were given 1,25- (OH) 2D3 (10ug/kg) every other day. The control group and the lack of the female rats were given 1ml saline instead of the stomach, until the twenty-first day after childbirth, again, again after the childbirth, again, again after the labor after the labor, again, again after the childbirth, again, again after the childbirth, again, after the labor, again, again after the labor after the twenty-first days, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again after delivery, again after delivery, again after childbirth, again, again after childbirth, again after childbirth, again, again after childbirth, again after childbirth, again after delivery, again after delivery, again after the birth, again after the birth, again, after the twenty-first day after childbirth, again The serum 25- (OH) D values were measured in three groups of female rats, and the serum 25- (OH) D values were measured at first days and 21 days after birth. The following indexes were observed: 1. mice weight and lung wet weight; 2. light microscopy was used to observe lung morphological structure; 3.q-PCR method was used to detect the expression of VEGF mRNA in lung tissue; 4. immunohistochemical methods were used. Do not detect the expression of VEGF and SP-C protein in lung tissue. Results: 1. the results of the model: feeding mice in different ways, after feeding 2 weeks, the serum 25- (OH) D value of the three groups of mice was not statistically significant (P0.05); the serum 25- (OH) D value of the lack of female mice was significantly lower than that of the control group and the intervention group, and there were significant differences (P0.01), intervention (P0.01). The serum 25- (OH) D value of the female rats was higher than that of the control group, the difference was statistically significant (P0.05); there was no significant difference in serum 25- (OH) D value between the first days of the newborn offspring of the newborn rats (P0.05), and the serum 25- (OH) D of the newborn rats was lower than that of the group and the intervention group (P0.05) in the twenty-first day of new birth (P0.05). The value of 25- (OH) D was slightly higher than that of the control group, but the difference was not statistically significant (P0.05) in.2. three groups of offspring rats weight and lung wet weight results: with the increase of age, the weight of the rats and the wet weight of lung were lower than those of the same age group, and the growth rate was slow, and the weight of 14,21 day offspring was less than that of the control group (P 0.05) the wet weight of lung in the 7,14,21 day rats was lighter than that of the control group (P0.01), and the weight of the rats in the intervention group was higher than that of the same age control group, and the growth rate was slightly faster than that of the control group (P0.05,0.01). The wet weight of the lungs in day 14,21 mice was significantly higher than that of the control group (P0.05).3. three groups. The results of HE staining light microscopy showed that the number of alveoli in the lung tissue was less than that of the same age group, the alveolus was small, the shape of the pulmonary alveolus increased and the interval of alveoli was wider, and the number of alveoli in the intervention group was slightly higher than that of the lung tissue in the control group, and the size of the alveolus was uniform and the solid structure area was reduced. The expression of VEGF mRNA in lung tissue of three groups of young rats was detected by.4.q-PCR method. The expression of VEGF mRNA in lung tissue of the control group and the intervention group increased gradually with the increase of day age. The first days after birth, the expression amount was higher than that of the control group and the intervention group. The difference was statistically significant (P0.05,0.01) and seventh days after birth. The rate of expression was significantly lower than that of the control group and the intervention group (P0.01), and the expression amount increased gradually, but the increase was slower than that of the control group and the intervention group. The growth curve was crossed with the normal group and the intervention group. The growth rate of the intervention group was faster than the control group. The expression amount was higher than that of the control group. The difference was statistically significant (P0.05,0.01).5. immunohistochemical method to detect the expression of VEGF protein in the lung tissue of three groups of offspring: with the increase of age, the expression of VEGF protein in the lung tissue of the control group and the intervention group increased gradually, and the expression of the first days after birth was slightly higher than that of the control group and the intervention group, seventh after birth, and seventh after birth. The difference between the control group and the intervention group was significantly lower than that of the control group and the intervention group (P0.05,0.01), and the expression amount increased gradually, and the growth amplitude was slower than that of the control group and the intervention group. The increase of the offspring rats in the intervention group was a little faster than the control group, and the expression amount was higher than that in the control group, and the difference was statistically significant (P). 0.05).6. immunohistochemical method was used to detect the expression of SP-C in lung tissue of three groups of offspring: the expression of SP-C in lung tissue increased gradually with the increase of day age in the three groups, and the expression of SP-C in lung tissue was lower than that of the same age group and the intervention group. The difference was statistically significant (P0.05,0.01). The expression of SP-C in lung tissue was higher than that of the same age control group, and there was a significant difference between the young rats at 14,21 day (P0.05). Conclusion: 1. mother rats during pregnancy, the lack of Vit D during lactation could inhibit the development of lung development of their offspring. If appropriate 1,25- (OH) 2D3 intervention was given, the lung development process of the offspring was certain to help.2. female rats and mammalian Vit D. Lack and proper supplement of 1,25- (OH) 2D3 may interfere with lung development by affecting the expression of VEGF and SP-C in the lung tissue of offspring rats.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.2
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