Ying Xie1, Xin Zhan2, Jiyuan Tu3, Kang Xu4, Xiongjie Sun5, Chunlian Liu6, Chang Ke7, Guosheng Cao8, Zhongshi Zhou9, Yanju Liu10. 1. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 18751028560@163.com. 2. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 872247547@qq.com. 3. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China; Hubei Research Center of Chinese Materia Medica Processing Engineering and Technology, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 3070@hbtcm.edu.cn. 4. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: kangxu05@hbtcm.edu.cn. 5. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 120143484@qq.com. 6. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 1624615167@qq.com. 7. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: kczzz@foxmail.com. 8. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: caoguosheng2006@163.com. 9. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: 3161@hbtcm.edu.cn. 10. School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China; Hubei Research Center of Chinese Materia Medica Processing Engineering and Technology, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, P.R. China. Electronic address: lyj1965954@hbtcm.edu.cn.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Atractylodes lancea (Thunb.) DC. is a widely used traditional herb that is well known for treating spleen deficiency and diarrhea. According to traditional Chinese medicine (TCM) theory, diarrhea-predominant irritable bowel syndrome (IBS-D) is caused by cold and dampness, resulting in diarrhea and abdominal pain. Nevertheless, the effect and mechanism of Atractylodes on IBS-D are still unclear. AIM OF THE STUDY: This study was designed to confirm the therapeutic effect of Atractylodes lanceolata oil (AO) in a rat model of IBS-D, and to determine the mechanisms by which AO protects against the disease. MATERIALS AND METHODS: The chemical components in AO were determined using gas chromatography-mass spectrometry (GC-MS). The expression levels of 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP), and surfactant protein (SP) in serum and colon tissue were measured using enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR), western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) were used to elucidate the mechanism of action of AO toward inflammation and the intestinal barrier in a rat model of IBS-D. RESULTS: The 15 chemical substances of the highest concentration in AO were identified using GC-MS. AO was effective against IBS-D in the rat model, in terms of increased body weight, diarrhea grade score, levels of interleukin-10 (IL-10), aquaporin 3 (AQP3), and aquaporin 8 (AQP8), and reduced fecal moisture content, levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), 5-HT, VIP, and SP, while also reducing intestinal injury, as observed using hematoxylin-eosin (HE) staining. In addition, the results indicated that AO increased the mRNA and protein expression levels of stem cell factor (SCF) and c-kit and enhanced the levels of zonula occludens-1 (ZO-1) and occludin, as well as decreased the levels of myosin light chain kinase (MLCK) and inhibited the phosphorylation of myosin light chain 2 (p-MLC2). CONCLUSIONS: AO was found to be efficacious in the rat model of IBS-D. AO inhibited the SCF/c-kit pathway, thereby reducing inflammation and protecting against intestinal barrier damage via the MLCK/MLC2 pathway.
ETHNOPHARMACOLOGICAL RELEVANCE: Atractylodes lancea (Thunb.) DC. is a widely used traditional herb that is well known for treating spleen deficiency and diarrhea. According to traditional Chinese medicine (TCM) theory, diarrhea-predominant irritable bowel syndrome (IBS-D) is caused by cold and dampness, resulting in diarrhea and abdominal pain. Nevertheless, the effect and mechanism of Atractylodes on IBS-D are still unclear. AIM OF THE STUDY: This study was designed to confirm the therapeutic effect of Atractylodes lanceolata oil (AO) in a rat model of IBS-D, and to determine the mechanisms by which AO protects against the disease. MATERIALS AND METHODS: The chemical components in AO were determined using gas chromatography-mass spectrometry (GC-MS). The expression levels of 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP), and surfactant protein (SP) in serum and colon tissue were measured using enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR), western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) were used to elucidate the mechanism of action of AO toward inflammation and the intestinal barrier in a rat model of IBS-D. RESULTS: The 15 chemical substances of the highest concentration in AO were identified using GC-MS. AO was effective against IBS-D in the rat model, in terms of increased body weight, diarrhea grade score, levels of interleukin-10 (IL-10), aquaporin 3 (AQP3), and aquaporin 8 (AQP8), and reduced fecal moisture content, levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), 5-HT, VIP, and SP, while also reducing intestinal injury, as observed using hematoxylin-eosin (HE) staining. In addition, the results indicated that AO increased the mRNA and protein expression levels of stem cell factor (SCF) and c-kit and enhanced the levels of zonula occludens-1 (ZO-1) and occludin, as well as decreased the levels of myosin light chain kinase (MLCK) and inhibited the phosphorylation of myosin light chain 2 (p-MLC2). CONCLUSIONS: AO was found to be efficacious in the rat model of IBS-D. AO inhibited the SCF/c-kit pathway, thereby reducing inflammation and protecting against intestinal barrier damage via the MLCK/MLC2 pathway.
Authors: Carrie L Kempf; Saad Sammani; Tadeo Bermudez; Jin H Song; Vivian Reyes Hernon; Matthew K Hufford; Jessica Burt; Sara M Camp; Steven M Dudek; Joe G N Garcia Journal: Pulm Circ Date: 2022-04-07 Impact factor: 2.886