电针足三里改善高脂血症小鼠的血脂紊乱：基于肠道菌群结构的改善

doi:10.12122/j.issn.1673-4254.2025.08.08

摘要/Abstract

摘要：

目的观察电针足三里对高脂饮食（HFD）诱导的高脂血症小鼠的疗效，探讨其作用机制。方法普通饮食C57BL/6J小鼠作为对照组（ND），高脂模型小鼠随机分为高脂饮食组（HFD组）、高脂饮食+电针足三里组（EA组），10只/组，EA组予电针足三里穴，记录3组小鼠在高脂饮食造模和针刺干预期间的体质量变化，干预16周后评估各组小鼠血脂4项（TC、TG、LDL-c、HDL-c）和血清丙二醛（MDA）含量；油红O染色观察肝脏形态学变化及脂质蓄积情况；Western blotting检测各组小鼠结肠组织中NLRP3、TLR4、IL-1b蛋白表达的影响；采用16S rRNA基因测序检测各组小鼠肠道菌群组成结构及相对丰度。结果干预16周，与HFD组比较，EA组小鼠体质量、血清TC、TG、LDL-C含量降低（P<0.01），MDA含量降低（P<0.05）；油红O染色结果表明，与ND相比，HFD组可见肝脏组织脂滴蓄积，脂滴明显升高，与HFD组相比，EA组小鼠肝脏的脂滴减少（P<0.05）；Western blotting显示与对照组相比，模型组小鼠结肠组织中NLRP3、TLR4、IL-1β蛋白表达均升高（P<0.05或P<0.01）；而与对照组相比，电针组小鼠结肠组织中NLRP3、TLR4、IL-1β蛋白表达差异无统计学意义（P>0.05）；与模型组相比，电针组小鼠结肠组织中NLRP3蛋白的表达降低（P<0.05）。16S rRNA测序表明EA治疗后肠道菌群数量明显回升（P<0.05），Muribaculaceae菌和Lachnospiraceae_NK4A136_group菌相对丰度升高（P<0.05）。结论电针足三里可能通过改善肠道菌群结构，增加Muribaculaceae属菌的丰度、促进降解高热量复合碳水化合物、改善胆固醇脂质代谢，并增加Lachnospiraceae_NK4A136_group菌而修复肠道黏膜、减轻毒素泄露，减少脂质过氧化物，减少肝脏脂肪沉积，降低高脂血症动物肠道黏膜NLRP3、TLR4、IL-1β的含量，减轻肠黏膜炎症，维护肠道屏障完整，改善血脂紊乱，为解释足三里的降脂作用提供了新的见解和实验依据。

关键词: 肠道菌群, 高脂血症, 足三里, 电针, NLRP3, TLR4, IL-1β

Abstract:

Objective To investigate the therapeutic effect of electroacupuncture (EA) at Zusanli (ST36) acupoint on hyperlipidemia in mice and explore the underlying mechanisms. Methods Thirty C57BL/6J mice were equally randomized into normal diet group, high-fat diet (HFD) group, and EA group. The changes in blood lipids and serum malondialdehyde (MDA) content of the mice were evaluated, and histopathological changes and lipid accumulation in the liver were observed using Oil red O staining (ORO). The expressions of NLRP3, TLR4, and IL-1β proteins in the colon tissues were detected with Western blotting, and gut microbiota changes were analyzed using 16S rDNA sequencing. Results In mice with HFD feeding, 16 weeks of EA treatment significantly lowered body weight and serum TC, TG, LDL-C and MDA levels, obviously reduced lipid accumulation in the liver, and ameliorated HFD-induced elevations of protein expressions of NLRP3, TLR4, and IL-1β. 16S rRNA sequencing revealed that EA significantly altered gut microbiota composition, and increased the diversity and relative abundance of beneficial bacterial groups such as Muribaculaceae and Lachnospiraceae NK4A136_group. Conclusion Electroacupuncture at ST36 alleviates blood lipid disorders in hyperlipidemic mice possibly by improving intestinal microbiota structure, promoting degradation of high-caloric carbohydrates, cholesterol lipid metabolism and intestinal mucosa repair, and reducing toxin leakage, lipid peroxides, and liver fat deposition.

Key words: gut microbiota, hyperlipidemia, Zusanli, electroacupuncture, NLRP3, TLR4, IL-1β

邓楚玉, 王雪莹, 甘立祥, 王大禹, 郑晓燕, 唐纯志. 电针足三里改善高脂血症小鼠的血脂紊乱：基于肠道菌群结构的改善[J]. 南方医科大学学报, 2025, 45(8): 1633-1642.

Chuyu DENG, Xueying WANG, Lixiang GAN, Dayu WANG, Xiaoyan ZHENG, Chunzhi TANG. Electroacupuncture at Zusanli improves blood lipid disorders in hyperlipidemic mice by improving gut microbiota structure[J]. Journal of Southern Medical University, 2025, 45(8): 1633-1642.

图/表 11

图1 电针足三里

Fig.1 Electroacupuncture at ST36 in a mouse.

图2 各组小鼠8周及16周体质量变化

Fig.2 Weight changes of the mice in each group at 8 and 16 weeks (Mean±SD, n=10). *P<0.05, **P<0.01vs ND, ##P<0.01 vs EA. ND:Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图3 各组小鼠血脂结果比较

Fig.3 Comparison of TC (A), TG (B), LDL-c (C) and HDL-c (D) levels of the mice among the groups (Mean±SD, n=6). *P<0.05, **P<0.01, ***P<0.001 vs ND, #P<0.05, ##P<0.01 vs EA. ND:Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图4 各组小鼠血清MDA比较

Fig.4 Comparison of serum MDA levels of the mice among the 3 groups (Mean±SD, n=6). ***P<0.001 vs ND, ##P<0.01 vs EA. ND: Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图5 电针足三里对高脂血症小鼠肝脏脂滴蓄积的影响

Fig.5 Lipid droplet accumulation in the liver of the mice (Oil Red Staining,Scale=1000 μm). A: ND group. B: HFD group. C: EA group. D: Comparison of Oil Red O stained area (%) among the 3 groups (Mean±SD, n=3). **P<0.01vs ND, #P<0.05 vs EA group. ND: Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图6 Western blotting分析小鼠结肠NLRP3、TLR4、IL-1b炎症相关蛋白表达

Fig.6 NLRP3, TLR4 and IL-1b protein expressions in mouse colon tissues detected by Western blotting (Mean±SD, n=4). A: Protein band of NLRP3, TLR4 and IL-1b. B-D: Bar chart of gray scale values of the protein bands. *P<0.05, **P<0.01vs ND group; #P<0.05 vs EA group. ND: Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图7 ASV 以及多样性指数稀释曲线

Fig.7 ASV and diversity index dilution curve in the 3 groups. ND: Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图8 ASV 多样性指数组间比较 Boxplot 图

Fig.8 Fig.8　Alpha diversity related boxplot. *P<0.05, **P<0.01. ND: Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图9 ASV花瓣图

Fig. 9 Flower plot. ND:Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图10 PCA分析

Fig.10 Principal component analysis. ND:Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

图11 菌群组成分析(属水平)

Fig.11 Bar chart of gut microbiota community structure at the genus level. ND:Normal diet group; HFD: High fat diet group; EA: High fat diet and electroacupuncture group.

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