EFFECT OF ISOLEUCINE ON mTOR SIGNALING PATHWAY AND NITROGEN METABOLISM OF CHINESE PERCH (SINIPERCA CHUATSI)
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摘要: 通过脑室注射异亮氨酸, 探究短期内异亮氨酸对鳜(Siniperca chuatsi)雷帕霉素靶蛋白(Mammalian target of rapamycin, mTOR)信号通路及氮代谢影响。结果显示: 脑室注射异亮氨酸后, (1)促进鳜氨氮排泄; (2)谷氨酸脱氢酶基因(Glutamate dehydrogenase, GDH)、谷草转氨酶基因(Glutamic oxaloacetic transaminase, GOT)和腺苷酸脱氨酶基因(Adenosine monophosphate deaminase, AMPD)氮代谢基因相对表达量显著性上调(P<0.05); (3)鳜血糖含量在0.5h显著性降低(P<0.05); (4)激活了鳜肝脏mTOR信号通路, 促使下游分子核糖体蛋白S6磷酸化(P<0.05)。结果表明: 异亮氨酸能够激活鳜肝脏mTOR信号通路, 介导氨基酸代谢, 提高鳜氮代谢基因的转录水平, 促使氨氮排泄增多。Abstract: In the present study, the effects of isoleucine on the mammalian target of rapamycin signaling (mTOR) pathway and nitrogen metabolism were investigated through Intraventricular (ICV) administration in Chinese perch (Siniperca chuatsi). In one hand, the ammonia excretion significantly increased, and mRNA levels of nitrogen metabolism genes involving glutamate dehydrogenase (GDH), glutamic oxaloacetic transaminase (GOT), and adenosine monophosphate deaminase (AMPD) remarkably increased after ICV injection of isoleucine as well (P<0.05). On the other hand, the results showed that blood glucose levels markedly decreased at 0.5h post-injection. The expression of liver ribosomal protein S6 was also notably enhanced, demonstrating that mTOR signaling pathway was activated. The results also indicated that isoleucine could activate the mTOR signaling pathway, mediate amino acid metabolism, and enhance ammonia-N excretion via nitrogen metabolism genes in Chinese perch.
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Keywords:
- Siniperca chuatsi /
- Isoleucine /
- Nitrogen metabolism /
- mTOR
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图 1 脑室注射异亮氨酸对鳜氨氮排泄的影响
值用平均数±标准误(n=6)来表示, 标有星号(*)表示注射异亮氨酸组与注射磷酸盐缓冲盐水的对照组相比有显著性差异(P<0.05); 下同
Figure 1. Effect of ICV injection of isoleucine on ammonia-N excretion of Chinese perch
Data are presented by mean±standard error of mean (SEM) (n=6). Significant level is marked with an asterisk (P<0.05), compared with the values obtained from phosphate buffered saline (PBS) injection into the muscle tissues of Chinese perch; the same applies below
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图 2 麦鲮氨氮排泄对水体氨氮浓度的影响
Figure 2. Effect of Cirrhinus mrigala on ammonia-N excretion on water (n=3)
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图 3 脑室注射异亮氨酸对鳜肝脏P-S6蛋白表达分析
Figure 3. The expression of liver ribosomal protein S6 of Chinese perch after ICV injection of isoleucine
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图 4 脑室注射异亮氨酸对鳜AMPD、GDH和GOT基因表达的影响
不同字母表示注射磷酸盐缓冲盐水组和异亮氨酸组各组之间分别在各个时刻有显著性差异(P<0.05)
Figure 4. The expression levels of AMPD, GDH and GOT of Chinese perch after ICV injection of isoleucine
Significant level is marked with different letters (P<0.05), compared with the values obtained from injections of isoleucine and PBS into the muscle tissues of Chinese perch, respectively
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图 5 脑室注射异亮氨酸对鳜血糖的影响
Figure 5. Effect of ICV injection of isoleucine on glucose level of Chinese perch
表 1 荧光定量PCR所用引物序列
Table 1 Primers used for real-time polymerase chain reaction (Real-Time PCR)
基因
Gene引物序列
Primer sequence (5′—3′)AMPD CATTTTCCTTCCCGTGTT
TCTGTCTGCGGAGTTGGTGDH GACGACGACCCCAACTTCT
GACCCGCTTCCTCTTCTGCGOT GGCTTTGCGAGTGGAGATA
GTGGGTTGGAGTAAATGGGRPL13A TATCCCCCCACCCTATGACA
ACGCCCAAGGAGAGCGAACT
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