Effect of Enteromorpha Polysaccharide on Intestinal Bacteria and Short Chain Fatty Acids in Obese Golden Hamsters
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摘要: 目的：探讨浒苔多糖对肥胖金黄地鼠血脂和肠道菌群构成的影响。方法：将40只雄性金黄地鼠按体重随机分为正常对照组（ND组）、高脂模型组（HFD组）、低剂量浒苔多糖组（LEP组，300 mg/kg·BW）和高剂量浒苔多糖组（HEP组，450 mg/kg·BW）。除对照组（ND 组）以普通饲料喂养外，其余组均以高脂饲料喂养。其中低、高剂量浒苔多糖组连续灌胃浒苔多糖水溶液。干预12周后，检测血脂水平，并采用16S rDNA高通量测序和气相色谱法分别比较各组间肠道菌群的多样性以及粪便中短链脂肪酸含量差异。结果：干预12周后，HFD组地鼠的体重、血清总胆固醇（Cholesterol，TC）、甘油三酯（Triglyceride，TG）和低密度脂蛋白胆固醇（Low density lipoprotein- cholesterol，LDL-C）显著高于ND组（P<0.05）。高剂量浒苔多糖可显著降低血清TC、TG、LDL-C和谷丙转氨酶（Alanine aminotransferase，ALT）水平（P<0.05）。16S rDNA高通量测序结果表明，在门水平上，HFD组厚壁菌门/拟杆菌门比例显著高于ND组（P<0.05）。与HFD组相比，HEP组的厚壁菌门/拟杆菌门比例显著降低（P<0.05）；在属水平上，HFD组的Eubacterium_coprostanoligenes_group（真杆菌属）、Lachnospiraceae_UCG-006（毛螺菌科 UCG-006）的相对丰度显著高于ND组（P<0.05），经高剂量浒苔多糖干预后，Eubacterium_coprostanoligenes_group（真杆菌属）、Lachnospiraceae_UCG-006（毛螺菌科 UCG-006）的相对丰度相对于HFD组显著降低（P<0.05）。此外，高脂饮食导致粪便中短链脂肪酸含量减少，高剂量浒苔多糖干预可显著增加粪便中短链脂肪酸含量（P<0.05）。结论：浒苔多糖可以通过调节高脂饲料喂养金黄地鼠肠道菌群构成以及短链脂肪酸生成，从而改善肥胖金黄地鼠的脂质代谢紊乱。Abstract: Objective: To investigate the effect of Enteromorpha polysaccharide (EP) on blood lipids and intestinal bacteria in obese golden hamsters. Method: Forty male golden hamsters were randomly divided into four groups, including control group (ND), model group (HFD), low-dose EP group (LEP, 300 mg/kg·BW), and high-dose EP group (HEP, 450 mg/kg·BW). The ND group was fed an ordinary diet, while the other three groups were given a high-fat diet. Among the high-fat diet groups, the LEP and HEP groups were continuously administered EP aqueous solution intragastrically. Twelve weeks later, the serum lipid levels were evaluated, the diversity and structural changes in the gut bacteria were examined using 16S rDNA sequencing, and the short-chain fatty acid concentration in faeces was examined using a gas chromatography flame ionisation detector (GC-FID). Results: After 12 weeks, the body weight, serum total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) of hamsters in the HFD group were considerably higher than those in the ND group (P<0.05). In contrast, high-dose EP treatment led to a significant decrease in serum TC, TG, LDL-C, and alanine aminotransferase (ALT) levels (P<0.05). Results of 16S rDNA sequencing revealed that at the phylum level the proportion of Firmicutes/Bacteroidetes was substantially higher in the HFD group than in the ND group (P<0.05). However, the level of Firmicutes/Bacteroidetes was much lower in the HEP group, compared to the HFD group (P<0.05). At the genus level, Eubacterium_coprostanoligenes_group and Lachnospiraceae UCG-006 were more abundant in the HFD group than in the ND group (P<0.05). Following high-dose EP intervention, the relative abundance of Eubacterium_coprostanoligenes_group and Lachnospiraceae_UCG-006 fell significantly, compared to the HFD group (P<0.05). Additionally, a high-fat diet feeding resulted in a decrease in the content of short-chain fatty acids in faeces, while high-dose EP intervention significantly increased the short-chain fatty acid content in faeces (P<0.05). Conclusion: The administration of EP alleviates the metabolic disorders of obese golden hamsters fed with a high-fat diet by moderating the composition of intestinal bacteria and enhancing the production of short-chain fatty acids.
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Key words:
- Enteromorpha polysaccharides /
- high-fat diet /
- gut microbiota /
- short-chain fatty acids
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图 2 门分类水平各组肠道微生物群落组成（A）以及差异菌门多重比较（B）
Figure 2. Intestinal microbial community composition of each group (A) and multiple comparison test (B) at phylum level
注：图（B）使用方差分析并通过Duncan法作多重比较 ；不同小写字母表示组间差异有统计学意义（P<0.05）；图3同。
图 3 属分类水平各组肠道微生物群落组成（A）以及多重比较（B）
Figure 3. Intestinal microbial community composition of each group (A) and multiple comparison test (B) at genus level
图 5 属分类水平差异肠道微生物物种与肥胖相关指标的相关性分析
Figure 5. Correlation analysis between gut microbiota at genus level and biochemical serum levels
注：*P<0.05，**P<0.01，***P<0.001表示有统计学差异；BW：体重，PTFC：睾周脂肪系数，PRFC：肾周脂肪系数；Higher in HFD表示该菌属在HFD组显著增加；Lower in HFD表示该菌属在HFD组显著减少。
图 6 菌群与短链脂肪酸的Spearman相关性分析
Figure 6. Spearman correlation analysis between gut microbiota and short-chain fatty acid
注：*P<0.05，**P<0.01，***P<0.001表示有统计学差异；Higher in HFD表示该菌属在HFD组显著增加；Lower in HFD表示该菌属在HFD组显著减少。
图 7 SCFAs与肥胖相关指标的Spearman相关性分析
Figure 7. Spearman correlation analysis between short-chain fatty acid and obesity-related parameters
注：*P<0.05，**P<0.01，***P<0.001表示有统计学差异；BW：体重，PTFC：睾周脂肪系数，PRFC：肾周脂肪系数；Lower in HFD表示在HFD组显著减少的短链脂肪酸；No sig表示在组间无统计学差异的短链脂肪酸。
表 1 浒苔多糖对金黄地鼠体重、脏器系数、血脂和肝功能的影响
Table 1. Effect of Enteromorpha polysaccharide on body weight, organ coefficient, blood lipid and liver function in golden hamster
分组 ND组 HFD组 LEP组 HEP组 体重（BW，g） 初始体重 129.28±6.12a 126.69±9.43a 128.07±6.53a 123.12±6.68a 最终体重 156.12±12.19b 174.71±13.96a 168.08±8.35ab 159.52±12.35b 脏器系数（g/100 g BW） 睾周脂肪系数 2.20±0.29b 3.07±0.36a 2.97±0.35a 2.47±0.38b 肾周脂肪系数 1.48±0.27b 2.35±0.39a 2.04±0.43a 1.69±0.26b 血脂参数（mmol/L） TC 4.55±0.37c 10.53±1.05a 8.91±1.20b 7.97±0.57b TG 1.10±0.21c 5.45±2.09a 5.10±1.02a 3.78±0.74b LDL-C 1.14±0.39c 4.60±1.05a 3.48±0.72b 2.84±0.37b HDL-C 3.61±0.49a 2.60±0.49b 2.79±0.53b 2.06±0.29c 肝功能（U/L） 血清ALT 19.23±6.12c 41.99±13.34a 28.30±10.45b 17.82±6.13c 血清AST 16.40±4.49a 15.16±4.84a 13.10±2.61a 12.77±1.41a 注：同行不同小写字母表示组间差异有统计学意义（P<0.05）；表2同。 
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表 2 浒苔多糖对金黄地鼠粪便短链脂肪酸含量的影响（mg/g）
Table 2. Effect of Enteromorpha polysaccharide on short-chain fatty acid content in feces of golden hamsters (mg/g)
分组 ND组 HFD组 HEP组 Acetic acid 1.433±0.440a 0.507±0.074b 0.594±0.136b Propanoic acid 0.275±0.149a 0.063±0.021b 0.143±0.067a Isobutyric acid 0.016±0.007a 0.011±0.006b 0.019±0.006a Butyric acid 0.363±0.178a 0.103±0.075b 0.214±0.138ab Isovaleric acid 0.034±0.013a 0.019±0.011b 0.036±0.011a Valeric acid 0.143±0.068a 0.020±0.008c 0.048±0.020bc Hexanoic acid 0.234±0.132a 0.017±0.002b 0b Heptylic acid 0.013±0.006a 0b 0b
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