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两种益生菌与枳实联用对功能性消化不良大鼠的改善作用及机制
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| 篇名： | 两种益生菌与枳实联用对功能性消化不良大鼠的改善作用及机制 |
| TITLE： | Ameliorative effects and mechanisms of two probiotics combined with Aurantii Fructus Immaturus on functional dyspepsia in rats |
| 摘要： | 目的 探讨两种益生菌（枯草芽孢杆菌、嗜酸乳杆菌）联合中药枳实改善大鼠功能性消化不良（FD）的作用及机制。方法将大鼠随机分为空白组、模型组、阳性对照组（多潘立酮，2.7mg/kg）、枳实组（9g/kg）、嗜酸乳杆菌组（5×107cfu/kg）、枯草芽孢杆菌组（5×107cfu/kg）、嗜酸乳杆菌+枳实组（嗜酸乳杆菌5×107cfu/kg+枳实9g/kg）、枯草芽孢杆菌+枳实组（枯草芽孢杆菌5×107cfu/kg+枳实9g/kg），每组8只。除空白组外，其余各组大鼠采用夹尾刺激+饥饱失常+游泳力竭应激干预法建立FD模型；成模后，各组大鼠灌胃相应药物/益生菌混悬液/生理盐水，每天1次，持续14d。末次给药后，检测大鼠胃排空率和小肠推进率；检测大鼠血清中脑肠肽相关指标[胃泌素（GAS）、P物质（SP）、血管活性肠肽（VIP）、生长抑素（SS）、胆囊收缩素（CCK）]水平；观察胃窦组织和十二指肠组织的病理学形态；收集大鼠盲肠内容物进行肠道菌群测序分析；检测大鼠胃窦组织中酪氨酸激酶受体c-Kit、干细胞因子（SCF）和十二指肠组织中Toll样受体4（TLR4）、髓样分化因子88（MyD88）、核因子κB（NF-κB）的蛋白表达水平。结果与空白组比较，模型组大鼠胃排空率、小肠推进率，血清中GAS、SP水平，厚壁菌门（Firmicutes）相对丰度，肠道菌群的Ace、Chao、Sobs指数，胃窦组织中SCF、c-Kit蛋白表达水平均显著降低（P＜0.05）；血清中VIP、SS、CCK水平，拟杆菌门（Bacteroidetes）相对丰度，十二指肠组织中TLR4、MyD88、NF-κB蛋白表达水平均显著升高（P＜0.05）；胃窦组织结构基本正常，但十二指肠组织结构存在异常，可见大量炎症细胞浸润。与模型组比较，各给药组大鼠上述指标水平大部分显著逆转（P＜0.05），胃窦组织结构正常；除枯草芽孢杆菌组、枯草芽孢杆菌+枳实组外，其余大鼠十二指肠病理状态均逐渐恢复。与各单药组相比，各联合组大鼠上述大部分指标进一步改善（P＜0.05）。结论枳实与上述两种益生菌联用均可改善FD大鼠的胃肠动力，且效果优于单独用药，具体作用机制可能与激活SCF/c-Kit信号通路、抑制TLR4/MyD88/NF-κB信号通路有关。 |
| ABSTRACT： | OBJECTIVE To investigate ameliorative effects and mechanisms of two probiotics （Bacillus subtilis， Lactobacillus acidophilus） combined with Aurantii Fructus Immaturus （AFI） on functional dyspepsia （FD） in rats. METHODS Rats were randomly divided into blank group， model group， positive control group （domperidone group， 2.7 mg/kg）， AFI group （9 g/kg）， L. acidophilus group （5×107 cfu/kg）， B. subtilis group （5×107 cfu/kg）， L. acidophilus+ AFI group （L. acidophilus 5×107 cfu/kg+ AFI 9 g/kg）， and B. subtilis+AFI group （B. subtilis 5×107 cfu/kg+AFI 9 g/kg）， with 8 rats in each group. Except for the blank group， FD model was established by tail-clamping stimulation+hunger and satiety disorder+swimming exhaustion in other groups. After modeling， each group was given the corresponding drug/probiotic suspensions/physiological saline intragastrically， once a day， for 14 consecutive days. After the last medication， gastric emptying rate and the rate of propulsion of the small intestine in rats were measured； the levels of brain-gut peptide-related indicators [gastrin （GAS）， substance P （SP）， vasoactive intestinal peptide （VIP）， somatostatin （SS） and cholecystokinin （CCK）] in the serum of rats were measured. The pathological morphology of the gastric antrum tissue and duodenal tissue was observed. Cecal contents from the rats were collected for gut microbiota sequencing analysis. The protein expression levels of tyrosine kinase receptor c-Kit and stem cell factor （SCF） in the gastric antrum tissue， as well as Toll-like receptor 4 （TLR4）， myeloid differentiation factor 88 （MyD88）， and nuclear factor κB （NF-κB） in the duodenal tissue of the rats were detected. RESULTS Compared with the blank group， model group showed significantly lower gastric emptying rate， small intestinal propulsion rate， serum levels of GAS and SP， relative abundance of Firmicutes， Ace， Chao and Sobs indexes of the gut microbiota， and protein levels of SCF and c-Kit in gastric antrum （P＜0.05）， while serum levels of VIP， SS and CCK， relative abundance of Bacteroidetes， as well as protein expressions of TLR4， MyD88， and NF-κB， were significantly higher （P＜0.05）. The histological structure JZYC23S53） of the gastric antrum tissue appeared basically normal； however， abnormalities were observed in the duodenal structure， with a significant infiltration of inflammatory cells visible. Compared with the model group， all treatment groups significantly modulated most of the above indexes （P＜0.05）. The histological structure of the gastric antrum tissue was normal. Except for the B. subtilis group and the B. subtilis+AFI group， the pathological states of the duodenum in the remaining rats gradually recovered. Compared with each single drug group， most of above indexes in rats from each combination group showed further improvement （P＜0.05）. CONCLUSIONS The combination of AFI with two probiotics can improve gastrointestinal motility in FD rats， and the effect is superior to that of using the drugs alone. The specific underlying mechanisms may be related to the activation of the SCF/c-Kit signaling pathway and the inhibition of the TLR4/MyD88/NF-κB signaling pathway. |
| 期刊： | 2025年第36卷第13期 |
| 作者： | 李宗年；熊英；龚晓惠；王兰兰；郭忠青；蒋林林；刘红樱；邓可众 |
| AUTHORS： | LI Zongnian， XIONG Ying，GONG Xiaohui，WANG Lanlan，GUO Zhongqing，JIANG Linlin，LIU Hongying，DENG Kezhong |
| 关键字： | 功能性消化不良；枳实；枯草芽孢杆菌；嗜酸乳杆菌；肠道菌群；SCF/c-Kit信号通路；TLR4/MyD88/NF-κB信号通路 |
| KEYWORDS： | functional dyspepsia； Aurantii Fructus Immaturus； Bacillus subtilis； Lactobacillus acidophilus； probiotics； SCF/c- |
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