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绞股蓝总皂苷对高胆固醇血症模型小鼠主要尿蛋白家族基因表达的影响研究
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| 篇名： | 绞股蓝总皂苷对高胆固醇血症模型小鼠主要尿蛋白家族基因表达的影响研究 |
| TITLE： | Study on the Effects of Gypenosides on Gene Expression of Major Urinary Proteins in Hypercholesterolemia Model Mice |
| 摘要： | 目的：探讨绞股蓝总皂苷（GPs）对高胆固醇血症模型小鼠肝组织中主要尿蛋白（Mups）基因表达的影响。方法：将C57BL/6J小鼠按体质量（BW）分为对照（ND）组、模型（HFD）组和GPs治疗（GP）组，每组11只。除ND组外，其余各组小鼠均给予高脂饲料以复制高胆固醇血症模型。于饲养的第17周起时，ND组和HFD组小鼠均灌胃等体积0.1％羧甲基纤维素钠溶液，GP组小鼠灌胃GPs混悬液（250mg/kg），每日1次，连续22周。在检测各组小鼠BW、血糖（BG）、血脂[总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）]水平的基础上，提取其肝组织总RNA，构建逆转录文库并进行RNA-seq测序；采用主成分分析（PCA）、火山图、散点图等方法筛选差异基因；采用实时荧光定量聚合酶链反应法（RT-qPCR）对差异基因进行验证；通过双变量分析评价差异基因表达量与上述药效学指标的相关性。结果：与ND组比较，HFD组小鼠BW和血清BG、TC、LDL-C水平均显著升高（P＜0.05）；与HFD组比较，GP组小鼠除BW外上述指标均显著降低（P＜0.05）。PCA结果显示，ND组和HFD组数据分布于不同象限，GP组数据分布介于上述两组之间。经高脂饮食诱导后，小鼠肝组织中Mup4、Mup5、Mup11、Mup15、Mup21基因mRNA均显著上调（P＜0.05）；经GPs干预后，Mup3、Mup4、Mup5、Mup8、Mup12、Mup21基因mRNA均显著下调（P＜0.05）。在ND组与HFD组、HFD组与GP组中表达发生显著改变且趋势相反的基因为Mup4、Mup5、Mup21。RT-qPCR验证结果显示，与ND组比较，HFD组小鼠Mup4、Mup5、Mup21基因mRNA的相对表达量均显著升高（P＜0.05）。相关性分析结果显示，小鼠Mup5基因mRNA表达量与血清TC、BG水平（r分别为0.7271、0.6706）以及Mup4、Mup21基因mRNA表达量与血清BG水平（r分别为0.7378、0.7215）均成正相关（P＜0.05）。结论：GPs对高胆固醇血症模型小鼠肝组织中Mups基因的表达具有一定的调控作用，可通过回调Mup4、Mup5、Mup21基因mRNA的过表达来降低模型小鼠的糖脂水平。 |
| ABSTRACT： | OBJECTIVE：To investigate the effects of gypenosides （GPs）on gene expression of major urinary proteins （Mups） in liver tissue of hypercholesterolemia model mice. METHODS ：C57BL/6J mice were divided into control （ND）group，model （HFD）group and GPs therapy （GP）group according to body weight （BW），with 11 mice in each group. Except for ND group ， other groups were given high-lipid diet to induce hypercholesterolemia model. From the 17th week of feeding ，ND group and HFD group were given constant volume of 0.1％CMC-Na solution intragastrically ；GP group were given GPs suspension （250 mg/kg） intragastrically，once a day ，for consecutive 22 weeks. BW ，the levels of blood glucose （BG）and blood lipid （TC，LDL-C）were detected in each group. Total RNA of liver tissue was extracted ，and reverse transcription library was constructed and RNA-seq sequencing was performed. The differentially expressed genes were screened by PCA ，volcano map and scatter plot. RT-qPCR was used for verification for differentially expressed genes. The correlation between the expression of differentially expressed genes and the above pharmacodynamic indexes was analyzed by bivariate analysis. RESULTS ：Compared with ND group ，BW，the levels of BG，TC and LDL-C in HFD group were increased significantly （P＜0.05）. Compared with HFD group ，above indexes of GP group were decreased significantly except for BW （P＜0.05）. PCA showed that the data of ND group and HFD group distributed in different quadrants ，and the data distribution of GP group was between above two groups. mRNA of Mup4，Mup5，Mup11，Mup15 and Mup21 in liver tissue of mice were increased significantly after treated with high-fat diet （P＜0.05）. mRNA of Mup3，Mup4， Mup5，Mup8，Mup12 and Mup21 were decreased significantly after treated with GPs （P＜0.05）. In ND group vs. HFD group and HFD group vs. GP group ，mRNA of Mup4，Mup5 and Mup21 genes changed significantly and the trend was opposite. Results of RT-qPCR verification showed that compared with ND group ，relative mRNA expression of Mup4，Mup5 and Mup21 gene were increased significantly in HFD group （P＜0.05）. Correlation analysis revealed that mRNA expression of Mup5 was positively correlated with the levels of TC and BG （r＝0.727 1，0.670 6，P＜0.05），mRNA expression of Mup4 and Mup21 were positively correlated with the level of BG （r＝0.737 8，0.721 5，P＜0.05）. CONCLUSIONS ：GPs can regulate the expression of Mups genes in liver tissue of hypercholesterolemia model mice ， and reduce glucose and lipid level through regulating the mRNA over-expression of Mup4，Mup5 and Mup21. |
| 期刊： | 2020年第31卷第15期 |
| 作者： | 杨艳萍，杜艺玫，秦琳，汪巍，鲁艳柳，陆安静，曾瑶，谭道鹏，何芋岐 |
| AUTHORS： | YANG Yanping ，DU Yimei，QIN Lin，WANG Wei，LU Yanliu ，LU Anjing ，ZENG Yao，TAN Daopeng ，HE Yuqi |
| 关键字： | 绞股蓝总皂苷；高胆固醇血症；主要尿蛋白；糖脂代谢；转录组学；小鼠 |
| KEYWORDS： | Gypenosides；Hypercholesterolemia；Major urinary proteins ；Glucose and lipid metabolism ；Transcriptome；Mice |
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