母鼠維生素D缺乏及干預(yù)對仔鼠肺發(fā)育及VEGF、SP-C表達影響的研究
發(fā)布時間：2018-07-26 07:58
【摘要】：目的:通過建立Vit D缺乏及Vit D干預(yù)母鼠模型,觀察母鼠孕期、哺乳期Vit D缺乏及適量1,25二羥維生素D3[1,25-(OH)2D3]干預(yù)對其仔鼠肺發(fā)育及肺血管內(nèi)皮生長因子(VEGF)、肺組織表面活性蛋白C(SP-C)的變化,從而探討母鼠Vit D缺乏及適量干預(yù)對仔鼠肺發(fā)育的影響及可能機制。方法:選取健康成年雌性清潔級SD大鼠,完全隨機分為正常對照組(對照組)、Vit D缺乏組(缺乏組)、1,25-(OH)2D3干預(yù)組(干預(yù)組),每組6只。對照組及干預(yù)組母鼠予以正常飼料、正常光照喂養(yǎng),缺乏組母鼠予不含Vit D飼料、避光喂養(yǎng),試喂2周后,用ELISA法測定三組母鼠血清25-(OH)D值,再與正常成年雄性SD大鼠合籠交配。于受孕第7天開始,干預(yù)組母鼠隔天一次給予1,25-(OH)2D3(10ug/kg)灌胃,對照組及缺乏組母鼠予1ml生理鹽水代替灌胃,一直到分娩后第21天,再次測三組母鼠血清25-(OH)D值,并檢測各組仔鼠生后第1天及第21天血清25-(OH)D值,再取生后第1、7、14、21天仔鼠肺組織,觀察以下指標:1.仔鼠體重及肺濕重;2.光鏡下觀察肺形態(tài)結(jié)構(gòu);3.q-PCR法檢測肺組織VEGF mRNA水平的表達;4.免疫組織化學(xué)方法分別檢測肺組織VEGF、SP-C蛋白表達。結(jié)果:1.造模結(jié)果:通過不同方式喂養(yǎng)母鼠,試喂2周后三組母鼠之間血清25-(OH)D值比較,差異無統(tǒng)計學(xué)意義(P0.05);分娩后第21天,缺乏組母鼠血清25-(OH)D值明顯低于對照組及干預(yù)組母鼠,有顯著差異(P0.01),干預(yù)組母鼠血清25-(OH)D值高于對照組母鼠,差異有統(tǒng)計學(xué)意義(P0.05);新生第1天仔鼠三組之間血清25-(OH)D值比較,差異無統(tǒng)計學(xué)意義(P0.05);新生第21天,缺乏組仔鼠血清25-(OH)D值低于對照組及干預(yù)組仔鼠,差異有統(tǒng)計學(xué)意義(P0.05),干預(yù)組仔鼠血清25-(OH)D值略高于對照組仔鼠,但差異無統(tǒng)計學(xué)意義(P0.05)。2.三組仔鼠體重及肺濕重結(jié)果:隨著日齡的增加,缺乏組仔鼠體重、肺濕重均低于同日齡對照組仔鼠,增長速度緩慢,缺乏組第14、21天仔鼠體重較對照組仔鼠輕,差異有統(tǒng)計學(xué)意義(P0.05),第7、14、21天仔鼠肺濕重較對照組仔鼠輕,有顯著差異(P0.01);干預(yù)組仔鼠體重、肺濕重均高于同日齡對照組仔鼠,增長速度稍快,其體重較對照組仔鼠比均有差異(P0.05,0.01),第14、21天仔鼠的肺濕重較對照組仔鼠比有差異(P0.05)。3.三組仔鼠HE染色光鏡觀察結(jié)果:缺乏組仔鼠較同日齡對照組仔鼠比肺組織肺泡數(shù)少,肺泡腔小,形態(tài)較不規(guī)則,實性團狀結(jié)構(gòu)區(qū)域增加,肺泡間隔較寬;干預(yù)組仔鼠較同日齡對照組仔鼠比肺組織肺泡數(shù)稍增加,肺泡腔大小均勻,實性團狀結(jié)構(gòu)區(qū)域減少,肺泡間隔較薄。4.q-PCR法檢測三組仔鼠肺組織VEGF mRNA表達結(jié)果:隨著日齡的增加,對照組及干預(yù)組仔鼠肺組織VEGF mRNA的表達量逐漸增多,缺乏組仔鼠生后第1天表達量高于對照組及干預(yù)組仔鼠,差異有統(tǒng)計學(xué)意義(P0.05,0.01),生后第7天較前下降,表達量明顯低于對照組及干預(yù)組仔鼠,有顯著差異(P0.01),之后表達量逐漸增加,但增長幅度較對照組及干預(yù)組仔鼠緩慢,增長曲線與正常組及干預(yù)組仔鼠有交叉現(xiàn)象;干預(yù)組仔鼠增長幅度較對照組仔鼠稍快,表達量均高于對照組仔鼠,差異有統(tǒng)計學(xué)意義(P0.05,0.01)。5.免疫組織化學(xué)方法檢測三組仔鼠肺組織VEGF蛋白表達結(jié)果:隨著日齡的增加,對照組及干預(yù)組仔鼠肺組織VEGF蛋白的表達量逐漸增多,缺乏組仔鼠生后第1天表達量較對照組及干預(yù)組仔鼠略高,生后第7天較前下降,低于對照組及干預(yù)組仔鼠,差異有統(tǒng)計學(xué)意義(P0.05,0.01),之后表達量逐漸增加,增長幅度較對照組及干預(yù)組仔鼠緩慢;干預(yù)組仔鼠增長幅度較對照組仔鼠稍快,表達量均高于對照組仔鼠,第14、21天仔鼠之間差異有統(tǒng)計學(xué)意義(P0.05)。6.免疫組織化學(xué)方法檢測三組仔鼠肺組織SP-C表達結(jié)果:三組仔鼠隨著日齡的增加,肺組織SP-C的表達量逐漸增加,第21天最高;缺乏組仔鼠肺組織SP-C表達量均低于同日齡對照組及干預(yù)組仔鼠,差異有統(tǒng)計學(xué)意義(P0.05,0.01);干預(yù)組仔鼠肺組織SP-C表達量較同日齡對照組仔鼠高,第14、21天仔鼠之間差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1.母鼠孕期、哺乳期Vit D缺乏會抑制其仔鼠生后的肺發(fā)育過程,若給予適量1,25-(OH)2D3干預(yù),對仔鼠的肺發(fā)育過程有一定幫助。2.母鼠孕期、哺乳期Vit D缺乏及適量補充1,25-(OH)2D3可能通過影響仔鼠肺組織VEGF、SP-C的表達,從而干擾其肺發(fā)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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R714.2
本文編號：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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R714.2
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