Alteration of Gene Expression Profile in Spontaneously Hypertensive Rats Treated with Protein Hydrolysate of Blue Mussel (Mytilus edulis) by DNA Microarray Analysis. Rattus norvegicus

The aim of this study was to investigate the antihypertensive effect of enzymatic hydrolysis of blue mussel protein (HBMP) in rats. Spontaneously hypertensive rats (SHRs) were orally administration with high- or low-dose of HBMP for 28 days. Major components of the renin-angiotensin (RAS) system in serum of SHRs from different groups were analyzed, and gene expression profiling were performed in the kidney of SHRs, using the Whole Rat Genome Oligonucleotide Microarray. Results indicated although genes involved in RAS system were not significantly altered, those related to blood coagulation system, cytokine and growth factor, and fatty acids metabolism were remarkablely changed. Several genes which were seldom reported to be implicated in pathogenesis of hypertension also showed significant expression alterations after oral administration of HBMP. Overall design: SHRs were randomly divided into three groups (n=10): control group (rats were orally administered with water, 3mL), low-dose group (rats were orally administered with HBMP, 10 mg/kg/day, 3mL), and high-dose group (rats were orally administered with HBMP, 20 mg/kg/day, 3mL). After orally administered with HBMP for 4 weeks, all rats were killed and the kidneys were dissected. For each group, equivalent amounts of RNA from four individual rats were mixed, and transcribed to Cy3-labeled cRNA using the Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies, Santa Clara, CA, USA). Then, Cy3-labeled cRNA from each group was hybridized to the Whole Rat Genome Oligonucleotide Microarray ver. 3.0 (4X44k, G2519F-028282) (Agilent Technologies), following the manufacturer's hybridization protocol.

本研究旨在探讨蓝贻贝蛋白酶解产物（HBMP）对大鼠的抗高血压作用。将自发性高血压大鼠（spontaneously hypertensive rats, SHRs）经口给予高剂量或低剂量的HBMP，持续干预28天。分析不同组别SHRs血清中肾素-血管紧张素系统（renin-angiotensin system, RAS）的主要组分，并采用全大鼠基因组寡核苷酸微阵列（Whole Rat Genome Oligonucleotide Microarray）对SHRs的肾脏组织开展基因表达谱分析。结果显示，尽管RAS系统相关基因未出现显著改变，但凝血系统、细胞因子与生长因子以及脂肪酸代谢相关基因均发生显著变化。另有若干此前鲜有报道与高血压发病机制相关的基因，在经口给予HBMP后也出现了显著的表达改变。实验整体设计：将SHRs随机分为3组（n=10）：对照组（经口给予去离子水，3mL）、低剂量组（经口给予HBMP，10mg/kg/天，3mL）以及高剂量组（经口给予HBMP，20mg/kg/天，3mL）。经口给予HBMP 4周后，处死所有大鼠并解剖获取肾脏组织。每组取4只个体大鼠的等量RNA进行混合，随后采用安捷伦低投入快速扩增标记试剂盒（Agilent Technologies，美国加利福尼亚州圣克拉拉市）将混合RNA反转录为经Cy3标记的cRNA。最后，按照制造商提供的杂交方案，将各组的Cy3标记cRNA与全大鼠基因组寡核苷酸微阵列版本3.0（4X44k，G2519F-028282，安捷伦科技）进行杂交。