DataSheet1_Effects of Maren Pills on the Intestinal Microflora and Short-Chain Fatty Acid Profile in Drug-Induced Slow Transit Constipation Model Rats.docx

Background: Slow transit constipation (STC) is becoming a common and frequently occurring disease in today’s society, and it is necessary to explore the safe and effective treatment of STC.Method: Our study aimed to investigate whether the laxative effect of Maren pills (MRW) is associated with the regulation of intestinal microflora and intestinal metabolism in the colon. Loperamide hydrochloride-induced STC rats received MRW intragastrically for two consecutive weeks to evaluate the laxative effect of MRW involving the regulation of intestinal microflora, intestinal metabolism, and 5-HT signaling pathway. Intestinal microflora was detected by 16s rDNA sequencing, intestinal metabolism of short-chain fatty acids (SCFAs) was detected by HPLC, and the 5-HT signaling pathway was detected by WB, ELISA, immunofluorescence, and immunohistochemical analysis.Results: Our results revealed that the treatments with MRW increased not only the body weight, 24-h fecal number, 24-h wet fecal weight, 24-h dry fecal weight, fecal water content, and the intestinal propulsion rate but also the colonic goblet cell number, colonic Muc-2 protein expression, and colonic mucus layer thickness in the STC model rats. Moreover, MRW activated the 5-HT pathway by increasing the levels of 5-HT, 5-HIAA, 5-HT4R, CFTR, cAMP, and PKA in the colon tissue of STC rats. The 16S rDNA sequencing results showed that MRW improved the colonic microflora structure in colonic contents of STC rats, mainly by increasing Lactobacillus and decreasing Prevotella. Finally, we found that MRW regulated the SCFA metabolism in the colonic contents of the STC rats, mainly by increasing the contents of acetic acid, propionic acid, and butyric acid; the relative abundance of Lactobacillus was positively correlated with either contents of acetic acid, propionic acid, and butyric acid, and the relative abundance of Clostridium was negatively correlated.Conclusion: Our study further showed that MRW could improve constipation in STC rats, and the mechanism may be by regulating the intestinal microflora structure and improving the metabolism of SCFAs.

背景：在现代社会中，慢传输性便秘（STC）已成为一种常见且频发的疾病，探索STC的安全有效治疗方法显得尤为必要。方法：本研究旨在探讨马仁丸（MRW）的通便作用是否与结肠中肠道微生态和肠道代谢的调节相关。通过给予洛哌丁胺诱导的STC大鼠MRW灌胃两周，评估MRW的通便效果，包括对肠道微生态、肠道代谢和5-HT信号通路的调节作用。肠道微生态通过16s rDNA测序检测，肠道短链脂肪酸（SCFAs）代谢通过高效液相色谱（HPLC）检测，5-HT信号通路通过蛋白质印迹（WB）、酶联免疫吸附测定（ELISA）、免疫荧光和免疫组织化学分析检测。结果：我们的研究结果揭示，MRW的治疗不仅增加了STC模型大鼠的体重、24小时粪便数量、24小时湿粪重、24小时干粪重、粪便含水量和肠道推进率，而且还增加了结肠杯状细胞数量、结肠Muc-2蛋白表达和结肠粘液层厚度。此外，MRW通过增加STC大鼠结肠组织中的5-HT、5-HIAA、5-HT4R、CFTR、cAMP和PKA水平，激活了5-HT通路。16S rDNA测序结果表明，MRW改善了STC大鼠结肠内容物中的结肠微生态结构，主要通过增加乳酸杆菌和减少普雷沃菌。最后，我们发现MRW调节了STC大鼠结肠内容物中的SCFA代谢，主要通过增加乙酸、丙酸和丁酸的含量；乳酸杆菌的相对丰度与乙酸、丙酸和丁酸的含量呈正相关，而梭菌属的相对丰度与这些含量呈负相关。结论：本研究进一步表明，MRW能够改善STC大鼠的便秘症状，其作用机制可能通过调节肠道微生态结构和改善短链脂肪酸的代谢来实现。