Therapeutic effect and mechanism by which Trichosanthis Fructus-Allii Macrostemonis Bulbus regulates gut microbiota in a rat model of coronary heart disease #br#

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doi:10.12307/2025.236

Abstract

Abstract: BACKGROUND: A network-based pharmacological approach has identified multifunctional effects of the main bioactive compounds in the Trichosanthis Fructus-Allii Macrostemonis Bulbus on coronary heart disease; however, the mechanism of its therapeutic effect on coronary heart disease has not been fully elucidated.
OBJECTIVE: To investigate the role and mechanism of Trichosanthis Fructus-Allii Macrostemonis Bulbus in improving coronary heart disease by regulating the composition of gut microbiota.
METHODS: Forty Sprague-Dawley rats were randomly divided into four groups: blank control group (n=10), model group (n=10), positive drug group (n=10), and medicine pair group (n=10). A rat model of coronary heart disease was established by continuous gastric perfusion of fat emulsion and injection of pituitrin. After modeling, rats in the model group were gavaged with distilled water (10 mL/kg) for control, rats in the positive drug group were gavaged with simvastatin 4 mg/kg per day, and rats in the medicine pair group were gavaged with Trichosanthis Fructus-Allii Macrostemonis Bulbus pairs 7.56 g/kg per day. All interventions lasted for 14 days. Electrocardiograms and myocardial pathology were observed, and blood lipid levels were measured. The structure of gut microbiota was analyzed using 16S rDNA sequencing technology.
RESULTS AND CONCLUSION: Electrocardiogram results showed ST segment elevation in the model group. There were no significant abnormalities in the electrocardiograms of the positive drug group and medicine pair group. Compared with the blank control group, the levels of total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol were significantly higher in the model group (P < 0.05). Compared with the model group, the levels of total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol were significantly lower in the positive drug group and medicine pair group (P < 0.05). Compared with the blank control group, focal myocardial cell necrosis was observed in the model group, while partial myocardial cell disarray was observed in the positive drug group and medicine pair group. Compared with the blank control group, the Ace, Shannon, and Chao indices were increased (P < 0.05) and the Simpson index was decreased (P < 0.05) in the model group, positive drug group and medicine pair group. Compared with the model group, the Ace and Chao indices were decreased (P < 0.05), while the Shannon index showed no significant difference (P > 0.05) and the Simpson index was also decreased (P < 0.05) in the positive drug group and medicine pair group. Compared with the blank control group, the relative abundances of Desulfovibrionia, Muribaculaceae_norank, etc. were increased in the model group, while those of Clostridia, [Eubacterium]_coprostanoligenes_group_norank, etc. were decreased. Compared with the model group, the relative abundances of WPS-2_norank, Muribaculaceae_norank, etc. were increased in the medicine pair group, while those of Clostridia, [Eubacterium]_coprostanoligenes_group_norank, etc. were decreased; the relative abundances of Desulfobacterota, [Eubacterium]_coprostanoligenes_group_norank, etc. were increased in the positive drug group, while those of Firmicutes, Muribaculaceae_norank, etc. were decreased. Compared with the positive drug group, the relative abundances of Desulfobacterota, Bacteroides, etc. were increased in the medicine pair group, while those of Firmicutes, [Eubacterium]_coprostanoligenes_group_norank, etc. were decreased. The LEfSe results showed that the medicine pair group had the highest microbial enrichment, followed by the blank control group and positive drug group, with the model group having the lowest microbial enrichment. To conclude, Trichosanthis Fructus-Allii Macrostemonis Bulbus pairs can improve the development of coronary heart disease by regulating gut microbiota composition, providing new insights for further research and development of Trichosanthis Fructus-Allii Macrostemonis Bulbus pairs.

Key words: Trichosanthis Fructus-Allii Macrostemonis Bulbus, 16S rDNA, coronary heart disease, gut microbiota, rat

CLC Number:

Sun Guanghan, Xie Zhencong, Sun Mi, Xu Yang, Guo Dong. Therapeutic effect and mechanism by which Trichosanthis Fructus-Allii Macrostemonis Bulbus regulates gut microbiota in a rat model of coronary heart disease #br#

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[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(5): 917-927.

Figures/Tables 20

胞浸润，细胞呈细长的圆柱形，横纹清晰，胞浆红染均质，细胞核圆形或椭圆形，各细胞形态较模型组排列整齐，边界清楚。见图2。 2.5 Alpha多样性分析各组稀释曲线逐渐趋于平缓，说明测序数据量合理，更多的数据量只会产生少量新的OUT，见图3。Alpha多样性主要评价肠道菌群丰富度和多样性。此次研究采用Ace指数、Shannon指数和Chao指数综合评价各组肠道菌群丰富度和多样性。分别对 Alpha diversity 的各个指数进行参数检验和非参数检验分析(样品比较则分别使用R中的 kruskal.test 函数和anova函数)，通过参数检验和非参数检验筛选不同条件下的显著差异的Alpha Diversity指数。与空白组比较，模型组、药对组的Ace、Shannon、Chao指数升高 (P < 0.05)，Simpson指数降低(P < 0.05)，见图4A；与空白组比较，模型组、阳性药组的Ace、Shannon、Chao指数升高(P < 0.05)，Simpson指数降低(P < 0.05)，见图4B。与模型组比较，阳性药组、药对组的Ace、Chao指数降低 (P < 0.05)，Shannon指数无差异(P > 0.05)，Simpson指数降低(P < 0.05)，见图4C。 2.6 Beta多样性分析通过分析不同样本OTU(97%相似性)组成可以反映样本间的差异和距离，PCA运用方差分解，将多组数据的差异反映在二维坐标图上，坐标轴取能够最大反映方差值的两个特征值。如样本组成越相似，反映在PCA图中的距离越近。 PCOA分析，用来研究样本群落组成的相似性或差异性，各组内样本相对聚集，表明组内差异较小，而各组间样本距离较远，表明各组间菌群构成具有明显差异，见图5。 2.7 肠道菌群落结构分析在分类学分析结果中，包含样本中含有何种微生物和各微生物的相对丰度。在门水平包括梭状芽孢杆菌门(Clostridia)、Bacilli、γ-变形菌门(Gammaproteobacteria)、Negativicutes、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)、疣微菌门(Verrucomicrobiota)、WPS-2、脱硫杆菌门(Desulfobacterota)、螺旋菌门(Spirochaetota)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)12种优势菌门。与空白组相比，模型组Desulfovibrionia、Spirochaetia、Gammaproteobacteria、Negativicutes、Bacteroidia、Proteobacteria相对丰度升高，Clostridia、Verrucomicrobiae、Bacilli、WPS-2_norank、Firmicutes、Actinobacteria相对丰度降低。与模型组相比，药对组WPS-2_norank、Desulfovibrionia、Actinobacteria、Spirochaetia、Bacteroidia、Bacilli相对丰度升高，Clostridia、Gammaproteobacteria、Negativicutes、Verrucomicrobiae相对丰度降低。与模型组相比，阳性药组Desulfobacterota、Spirochaetota、Verrucomicrobiota、Bacteroidota相对丰度升高，Firmicutes、Proteobacteria、WPS-2相对丰度降低。与阳性药相比，药对组Desulfobacterota、Actinobacteriota、Proteobacteria、Spirochaetota、Bacteroidota相对丰度升高，Firmicutes、Verrucomicrobiota相对丰度降低。见图6。粪杆菌真核菌群([Eubacterium]_coprostanoligenes_group)、Muribaculaceae_norank、拟杆菌属(Bacteroides)、未分类的毛螺菌属(Lachnospiraceae_unclassified)、 Ruminococcus、Clostridia_UCG-014_norank(相对丰度值> 4%)为空白组肠道菌群在属水平上的主要构成。与空白组相比，模型组Muribaculaceae_norank相对丰度升高，[Eubacterium]_coprostanoligenes_gro成up_norank、Bacteroides相对丰度降低。与模型组相比，药对组Muribaculaceae_norank、Bacteroides相对丰度升高，[Eubacterium]_coprostanoligenes_group_norank相对丰度降低。与模型组相比，阳性药组[Eubacterium]_coprostanoligenes_group_norank相对丰度升高，Muribaculaceae_norank相对丰度降低。与阳性药组相比，药对组Bacteroides、Clostridia_UCG-014_norank、Muribaculaceae_norank相对丰度升高，[Eubacterium]_coprostanoligenes_group_norank、Ruminococcus、Lachnospiraceae_unclassified相对丰度降低，见图7。 2.8 肠道菌群组间差异分析 LEfSe采用线性判别分析(LDA)来估算每个组分(物种)丰度对差异效果影响的大小。结果发现：共29条差异显著的种群(LDA值> 4)，其中12条富集于空白组，15条富集于药对组，2条富集于模型组，见图8A；共16条差异显著的种群(LDA值> 4)，其中10条富集于空白组，7条富集于阳性药组，4条富集于模型组，见图8B；共18条差异显著的种群(LDA值> 4)，其中6条富集于阳性药组，10条富集于药对组，2条富集于模型组，见图8C。"

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