参附注射液对异丙肾上腺素诱导的慢性心衰大鼠肠道菌群及肠道通透性的影响

作者：张 倩，周 颖，秦思佳，伍梅芳

单位：长沙卫生职业学院，湖南 长沙 410605

引用：引用：张倩，周颖，秦思佳，伍梅芳.参附注射液对异丙肾上腺素诱导的慢性心衰大鼠肠道菌群及肠道通透性的影响[J].中医药导报,2025,31(12):67-74.

DOI：10.13862/j.cn43-1446/r.2025.12.011

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摘要：目的：探究参附注射液对异丙肾上腺素（ISO）诱导的慢性心衰大鼠肠道菌群及肠道通透性的影响。方法：将32只SPF级雄性SD大鼠随机分为正常组（CONT组，6只）和模型制备组（26只）。模型制备组大鼠皮下注射ISO（5 mg/kg），CONT组大鼠注射等量生理盐水，连续给药14 d。完成模型制备后，将符合慢性心力衰竭标准的大鼠随机分为模型组（MOD组）、参附组（SF组）和卡托普利组（KT组），分别给予相应干预措施，持续15 d。干预后采用心脏彩超检测大鼠心功能，称量大鼠体质量、心脏质量、左心室质量并计算心脏质量指数、左心室质量指数；心脏苏木精-伊红染色（HE染色）观察心肌组织结构改变情况；酶联免疫吸附试验（ELISA）检测血清氨基末端B型脑钠肽前体（NT-proBNP）含量；蛋白质印迹法（Western blotting）检测小肠组织中闭锁小带蛋白1（ZO-1）、闭合蛋白（occludin）表达水平；采用RT-qPCR技术检测小肠组织中ZO-1 mRNA和occludin mRNA表达水平；16S rDNA检测肠道内容物菌群。结果：MOD组大鼠左室射血分数（LVEF）、左室短轴缩短率（LVFS）及体质量低于CONT组（P＜0.01），LVEDD、LVESD、心脏质量指数、左心室质量指数及血清NT-proBNP含量高于CONT组（P＜0.01）；SF组、KT组大鼠LVEF、LVFS、体质量高于MOD组（P＜0.01或P＜0.05），LVESD、心脏质量指数、左心室质量指数及血清NT-proBNP含量均低于MOD组（P＜0.01或P＜0.05）。心脏HE染色提示CONT组大鼠心肌细胞排列规则，结构完整；MOD组大鼠心肌呈现显著病变，心肌细胞结构紊乱，部分心肌纤维断裂、熔解，炎症细胞浸润；SF组大鼠心肌细胞间质内纤维结缔组织和炎症细胞较MOD组明显减少，心肌细胞结构完整，排列整齐；KT组大鼠心肌纤维排列较整齐，炎症浸润显著减轻。MOD组大鼠小肠组织ZO-1及occludin蛋白相对表达量低于CONT组（P＜0.01）；SF组、KT组大鼠小肠组织中ZO-1、occludin蛋白相对表达量均高于MOD组（P＜0.01）。MOD组大鼠小肠组织ZO-1 mRNA及occludin mRNA相对表达量低于CONT组（P＜0.01）；SF组、KT组大鼠小肠组织ZO-1mRNA和occludin mRNA相对表达量均高于MOD组（P＜0.01）。16S rDNA检测结果显示，MOD组大鼠Shannon指数、Ace指数及Chao1指数低于CONT组（P＜0.05），Simpson指数高于CONT组（P＜0.01）；SF组、KT组大鼠Shannon指数、Chao1指数均高于MOD组（P＜0.05或P＜0.01），Simpson指数低于MOD组（P＜0.05或P＜0.01）。β多样性结果显示，SF组大鼠肠道菌群分布更接近CONT组；MOD组大鼠乳酸杆菌属（Lactobacillus）的相对丰度低于CONT组（P＜0.05），狭义梭菌属_1（Clostridium_sensu_stricto_1）及unclassified_f_Peptostreptococcaceae相对丰度高于CONT组（P＜0.01）；SF组大鼠乳酸杆菌属的相对丰度高于MOD组（P＜0.01），狭义梭菌属_1及unclassified_f_Peptostreptococcaceae相对丰度低于MOD组（P＜0.01）。结论：参附注射液能提高慢性心衰大鼠的心功能，恢复大鼠的肠道通透性，改善肠道菌群紊乱。

关键词：慢性心衰；参附注射液；肠道菌群；肠道通透性；大鼠

Abstract：

Objective: To investigate the effects of Shenfu injection (SF) on intestinal microbiota and intestinal permeability in rats with chronic heart failure (CHF) induced by isoproterenol (ISO). Method: Totally 32 SPF-grade male Sprague-Dawley (SD) rats were randomly divided into control group (CONT group, n=6) and model preparation group (n=26). The model preparation group was subcutaneously injected with ISO (5 mg/kg) for 14 consecutive days to establish the CHF model, while the CONT group received subcutaneous injections of saline for 14 days. After modeling, the CHF rats were randomly divided into model group (MOD group), Shenfu injection group (SF group), and captopril group (KT group), with each group receiving corresponding interventions for 15 days. After the intervention, cardiac ultrasound was used to assess cardiac function in rats. The body weight, heart weight and left ventricular weight of the rats were measured, and the heart mass index and left ventricular mass index were calculated. Myocardial tissue structural changes were observed through hematoxylin and eosin staining (HE staining) of the heart. Serum levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP) were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of zonula occludens-1 (ZO-1) and occludin in intestinal tissue were detected by Western blotting. RT-qPCR was employed to measure the expression levels of ZO-1 mRNA and occludin mRNA in intestinal tissue. The gut microbiota in intestinal contents was analyzed using 16S rDNA sequencing. Results: In rats of the MOD group, the left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), and body weight were lower than those in the CONT group (P<0.01), while the left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), cardiac mass index, left ventricular mass index, and serum level of NT-proBNP were higher than those in the CONT group (P<0.01). In rats of the SF group and KT group, the LVEF, LVFS and body weight were higher than those in the MOD group (P<0.01 or P<0.05), while the LVESD, cardiac mass index, left ventricular mass index and serum level of NT-proBNP were lower than those in the MOD group (P<0.01 or P<0.05). HE staining revealed that cardiomyocytes in the CONT group were regularly arranged with intact structure, and that the MOD group exhibited significant myocardial pathological changes, including disordered cardiomyocyte structure, fragmentation and lysis of some myocardial fibers, and inflammatory cell infiltration. In the SF group, the fibrous connective tissue and inflammatory cells in the myocardial interstitium were significantly reduced compared with the MOD group, and cardiomyocytes remained intact and regularly arranged. In the KT group, myocardial fibers were relatively regularly arranged, and inflammatory infiltration was significantly alleviated. In the MOD group, the relative expression levels of ZO-1 and occludin proteins in the small intestinal tissue of rats were lower than those in the CONT group (P<0.01). In the SF group and KT group, the relative expression levels of ZO-1 and occludin proteins in the small intestinal tissue of rats were higher than those in the MOD group (P<0.01). In the MOD group, the relative expression levels of ZO-1 mRNA and occludin mRNA in the small intestinal tissue of rats were lower than those in the CONT group (P<0.01). In the SF group and KT group, the relative expression levels of ZO-1 mRNA and occludin mRNA in the small intestinal tissue of rats were higher than those in the MOD group (P<0.01). 16S rDNA sequencing results revealed that the Shannon index, Ace index, and Chao1 index of rats in the MOD group were lower than those in the CONT group (P<0.05), while the Simpson index was higher than that in the CONT group (P<0.01). The Shannon index and Chao1 index of rats in the SF group and KT group were higher than those in the MOD group (P<0.05 or P<0.01), while the Simpson index was lower than that in the MOD group (P<0.05 or P<0.01). Results of β-diversity analysis showed that the intestinal microbiota distribution of the SF group was closer to that of the CONT group. In the MOD group, the relative abundance of Lactobacillus in rats was lower than that in the CONT group (P<0.05), while the relative abundances of Clostridium_sensu_stricto_1 and unclassified_f_Peptostreptococcaceae were higher than those in the CONT group (P<0.01). In the SF group, the relative abundance of Lactobacillus in rats was higher than that in the MOD group (P<0.01), while the relative abundances of Clostridium_sensu_stricto_1 and unclassified_f_Peptostreptococcaceae were lower than those in the MOD group (P<0.01). Conclusion: Shenfu injection can improve cardiac function in rats with chronic heart failure, restore intestinal permeability, and ameliorate gut microbiota dysbiosis.

Key words：chronic heart failure; Shenfu injection; intestinal microbiota; intestinal permeability; rat

发布时间：2025-12-31

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