何首乌对高血脂小鼠的降脂作用及转录组学研究

Hypolipidemic Effect and Transcriptome of Fallopia multiflora(Thunb.)Harald.on Hyperlipidemic Mice

研究何首乌对高血脂小鼠血清的影响,并探讨何首乌治疗高血脂症小鼠肝脏的转录组变化。60只昆明小鼠随机分为5组:正常对照组、高血脂模型组和高、中及低浓度何首乌提取液组。除正常对照组外,其他4组采用高脂饲料喂养,诱导建立高血脂模型。8周后高、中及低浓度何首乌提取液组分别采用1.0 g/mL、0.6 g/mL和0.3 g/mL腹腔注射(0.1 mL/10 g)。正常对照组,高血脂模型组腹腔注射等体积生理盐水,每天给药1次,持续60 d。摘眼球取血,ELISA法检测血清TC、TG、LDL-C和HDL-C水平。采用RNAseq测序方法检测小鼠肝组织基因转录水平。生物信息学方法分析各组样本间差异表达基因,并通过实时荧光定量PCR验证部分基因的表达。与模型对照组相比,何首乌提取液高浓度组TC、TG和LDL-C水平显著降低,而HDL-C水平明显升高(P<0.05)。转录组测序共检测到787个显著差异表达的基因,其中上调基因260个,下调基因527个。GO分析表明,差异表达基因富含了532种细胞组分类别,参与了933个分子功能和2869个生物过程。KEGG富集通路分析表明,何首乌降脂过程参与多条脂质代谢通路,如脂肪酸代谢、亚油酸代谢和花生四烯酸代谢等。转录因子分析共发现67个具有显著差异的转录因子。高浓度何首乌提取液对高血脂模型小鼠有明显的降脂作用,其降脂作用可能是通过调控脂质代谢相关基因表达实现的。

To investigate the effect and mechanism of extraction of Fallopia multiflora(Thunb.)Harald.On serum of hyperlipidemic mice models.60 Kunming mice were randomly divided into 5 groups,normal control group,hyperlipidemic model group,high,medium,and low concentration of F.multiflora groups.In addition to the normal control group,the other 4 groups were fed with high-fat diet and induced to establish the hyperlipidemic model.After 8 weeks,the mice in the high,mediumand low concentration groups were injected intraperitoneally with 1.0 g/mL,0.6 g/mL and 0.3 g/mL extraction of F.multiflora(0.1mL/10g)respectively.Mice in the normal control and hyperlipidemic model groups were injected with 0.9%physiological saline.The treatment was given once a day for 60 days.The levels of TC,TG,LDL-C and HDL-C were detected by ELISA kits.The RNA-seq sequencing method was used to detect the gene transcription level in mice liver tissues.Bioinformatics method was used to analyze the differentially expressed genes among samples,the expression of several genes was verified by the real-time fluorescent quantitative PCR.Compared with the hyperlipemia model control group,the level of TC,TG and LDL-C decreased significantly in the high concentration of F.multiflora.group(P<0.05).While the level of HDL-C increased significantly(P<0.05).There were 787 differentially expressed genes,including 260 up-regulated genes and 527 down-regulated genes.Differentially expressed genes are riched in 532 cellular components,involved in 933 molecular functions and 2869 biological processes.The KEGG enrichment pathway showed that F.multiflora could improve the hyperlipemia through several lipid metabolic pathways including fatty acid degradation,linoleic acid metabolism,and arachidonic acid metabolism pathways.A total of 67 transcription factors with significant pro-portional difference were found by transcription factor analysis.The high concentration F.multiflora of has obvious hypolipidemic effect on hyperlipidemia model mice.And the hypolipidemic effect may be realized by regulating the expression of genes related to lipid metabolism.