饲喂高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响

doi:10.3969/j.issn.1004-1524.2021.05.04

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (5): 801-808.DOI: 10.3969/j.issn.1004-1524.2021.05.04

饲喂高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响

王志(), 胡仲皓, 桂雪儿, 冯士彬, 李玉, 王希春, 李锦春, 吴金节^*()

安徽农业大学动物科技学院,安徽合肥 230036

收稿日期:2020-07-10 出版日期:2021-05-25 发布日期:2021-05-25
通讯作者: 吴金节
作者简介:*吴金节,E-mail:wjj@ahau.edu.cn
王志(1994—),女,安徽安庆人,硕士研究生,从事畜禽营养代谢病研究。E-mail:3034717518@qq.com
基金资助:
国家重点研发计划项目(2016YFD0501205);安徽省重点研究与开发计划项目(1804a07020135);安徽农业大学研究生创新基金

Effects of high protein diets on serum uric acid levels, liver and kidney ultrastructure and expression of ABCG2 in goslings

WANG Zhi(), HU Zhonghao, GUI Xueer, FENG Shibin, LI Yu, WANG Xichun, LI Jinchun, WU Jinjie^*()

College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

Received:2020-07-10 Online:2021-05-25 Published:2021-05-25
Contact: WU Jinjie

摘要/Abstract

摘要：

通过建立雏鹅高尿酸血症模型,研究高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响。超微结构观察发现,对照组(CP)肝脏和肾脏细胞器结构完好,高蛋白1组(HP1)、高蛋白2组(HP2)肝脏部分线粒体开始出现肿胀、空泡变性,内质网肿胀消失,糖原减少。肾脏细胞部分线粒体开始出现肿胀、空泡变性,核膜间隙增大。雏鹅7日龄时,HP2组鹅的血清尿酸水平极显著高于CP组和HP1组(P<0.01);雏鹅14日龄时,HP2组鹅的血清尿酸水平极显著高于CP组和HP1组(P<0.01),HP1组鹅血清尿酸水平极显著高于CP组(P<0.01)。qPCR、Western-boltting及免疫组化结果显示,HP2组鹅十二指肠、空肠中ABCG2基因和蛋白的表达显著或极显著高于CP组和HP1组(P<0.05或P<0.01),鹅肾脏ABCG2基因和蛋白的表达显著或极显著低于CP组和HP1组(P<0.05或P<0.01);HP1组鹅ABCG2基因和蛋白在空肠中的表达显著高于CP组(P<0.05),在肾脏中的表达显著低于CP组(P<0.05)。综上所述,饲喂高蛋白日粮导致雏鹅肝脏、肾脏细胞受损,血清尿酸水平升高,引发雏鹅高尿酸血症,空肠、十二指肠中ABCG2的表达增加,而肾脏中ABCG2的表达降低。

关键词: 高蛋白日粮, 高尿酸血症, ABCG2, 雏鹅

Abstract:

In this study, the hyperuricemia model of gosling was established to study the effects of high protein diets on serum uric acid level, ultrastructure of liver and kidney and expression of ABCG2 in goslings.Transmission electron microscopy showed that the organelles structure of gosling liver and kidney in the control group (CP) was complete, whereas, some mitochondria of liver in high protein group 1 (HP1) and high protein group 2 (HP2) appeared to be swollen and showed vacuole degeneration, the endoplasmic reticulum was dilated, and glycogen levels were decreased. Some mitochondria of kidney cells appeared to be swollen, showed vacuole degeneration, and the nuclear membrane space increased. At 7 days of age, the serum uric acid levels in HP2 group were significantly higher than those of CP and HP1 groups (P<0.01). At 14 days of age, the serum uric acid level in HP2 group was significantly higher than those of CP and HP1 groups (P<0.01), and the serum uric acid level in HP1 group was significantly higher than those of CP group (P<0.01).The results of qPCR, Western-boltting and immunohistochemistry showed that, the expression of ABCG2 gene and protein in duodenum and jejunum in HP1 group was significantly higher than that in CP group and HP1 group (P<0.05 or P<0.01), and the expression in kidney was significantly lower than that in CP group and HP1 group (P<0.05 or P<0.01). The expression of ABCG2 gene and protein in HP1 group was significantly higher than that in CP group (P<0.05), and the expression in kidney was significantly lower than that in CP group (P<0.05). In conclusion, feeding high protein diet resulted in the damage of liver and kidney cells, the increase of uric acid level in vivo, the hyperuricemia of gosling, the increase of ABCG2 expression in jejunum and duodenum, and the decrease of ABCG2 expression in kidney.

Key words: high protein diet, hyperuricemia, ABCG2, gosling

中图分类号:

S835

王志, 胡仲皓, 桂雪儿, 冯士彬, 李玉, 王希春, 李锦春, 吴金节. 饲喂高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响[J]. 浙江农业学报, 2021, 33(5): 801-808.

WANG Zhi, HU Zhonghao, GUI Xueer, FENG Shibin, LI Yu, WANG Xichun, LI Jinchun, WU Jinjie. Effects of high protein diets on serum uric acid levels, liver and kidney ultrastructure and expression of ABCG2 in goslings[J]. Acta Agriculturae Zhejiangensis, 2021, 33(5): 801-808.

图/表 7

表1 鹅的饲粮营养组成与含量(风干基础)

Table 1 Composition and nutrient levels of diets in goslings (air-dry basis)

项目 Items	含量 Content
项目 Items	对照组 CP		高蛋白1组 HP1		高蛋白2组 HP2
原料 Ingredients
玉米 Corn/%		32.52		30.33		19.52
豆粕 Soybean meal/%		—		21.39		13.70
玉米蛋白粉		6.06		9.40		13.30
Corn protein meal/%
花生粕 Peanut meal/%		8.64		—		14.60
青糠 Green bran/%		24.46		18.39		18.39
次粉 Wheat middling/%		15.23		9.67		9.67
酒糟蛋白饲料 DDGS/%		7.04		4.77		4.77
石粉 Limestone/%		2.10		2.10		2.10
磷酸氢钙 CaHPO₄/%		1.07		1.07		1.07
预混料 Premix/%		2.88		2.88		2.88
合计 Total/%		100		100		100
营养水平 Nutrient levels
粗蛋白 Crude protein/%		18		23		28
代谢能 ME/(MJ·kg^-1)		2.76		2.72		2.7
钙 Ca/%		1.05		1.05		1.05
总磷 TP/%		0.58		0.58		0.58
有效磷 AP/%		0.35		0.35		0.35
赖氨酸 Lys/%		0.81		1.26		1.26
蛋氨酸 Met/%		0.30		0.42		0.46
蛋氨酸+半胱氨酸		0.58		0.81		0.88
Met+Cys/%

图1 各组鹅肝脏细胞电镜图片放大倍数20 000×。A,CP组;B,HP1组;C,HP2组。下同。

Fig.1 Transmission electron microscopic pictures of liver cells of goslings in each group The magnification was 20 000 times the original size. A, Group CP; B, Group HP1; C, Group HP2. The same as below.

图2 各组鹅肾脏细胞电镜图片

Fig.2 Transmission electron microscopic pictures of kidney cells of goslings in each group

表2 7日龄和14日龄雏鹅血清尿酸值

Table 2 Serum uric acid values of 7 and 14 day old goslings

处理 Treatment	血清尿酸值/(μmol·L^-1)
处理 Treatment	第7天 7th day	第14天 14th day
对照组CP	157.40±21.08 B	207.14±19.46 C
高蛋白1组HP1	201.72±27.66 B	452.58±60.26 B
高蛋白2组HP2	271.26±48.62 A	810.38±103.85 A

图3 鹅肝脏、肾脏、十二指肠、空肠中ABCG2-mRNA相对表达量同一组织不同处理间没有相同小写字母表示差异显著(P<0.05),没有相同大写字母表示差异极显著(P<0.01)。下同。

Fig.3 Relative expression level of ABCG2-mRNA in liver, kidney, duodenum and jejunum of goslings The bars without the same lowercase or uppercase letters within the same organ showed significant difference at the level of 0.05 or 0.01, respectively. The same as below.

图4 鹅肝脏、肾脏、十二指肠、空肠中ABCG2蛋白表达量

Fig.4 ABCG2 expression in liver, kidney, duodenum and jejunum of goslings

图5 鹅肝脏、肾脏、十二指肠、空肠中ABCG2蛋白表达量

Fig.5 ABCG2 protein expression in liver, kidney, duodenum and jejunum of goslings

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饲喂高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响

Effects of high protein diets on serum uric acid levels, liver and kidney ultrastructure and expression of ABCG2 in goslings

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