The Traditional Medicine Banhasasim-Tang Depolarizes Pacemaker Potentials of Cultured Interstitial Cells of Cajal through M3 Muscarinic and 5-HT3 Receptors in Murine Small Intestine

Abstract

Background: Banhasasim-tang (BHSST) is a classic herbal formulation in traditional Chinese medicine widely used for gastrointestinal (GI) tract motility disorder. We investigated the effects of BHSST on the pacemaker potentials of cultured interstitial cells of Cajal (ICCs) in small intestine in vitro and its effects on GI motor functions in vivo. Methods: We isolated ICCs from the small intestines and recorded pacemaker potentials in cultured ICCs with the whole-cell patch-clamp configuration in vitro. Intestinal transit rates (ITR%) were investigated in normal mice and GI motility dysfunction (GMD) mouse models in vivo. Results: BHSST (20-50 mg/mL) depolarized pacemaker potentials and decreased their amplitudes in a concentration-dependent manner. Pretreatment with methoctramine (a muscarinic M 2 receptor antagonist) did not inhibit BHSST-induced pacemaker potential depolarization. However, when we applied 1,1-dimethyl-4-diphen-ylacetoxypiperidinium iodide (4-DAMP; a muscarinic M 3 receptor antagonist), BHSST-induced effects were blocked. Pretreatment with Y25130 (a 5-HT 3 receptor antagonist) blocked BHSST-induced effects in ICCs. In addition, when we applied 4-DAMP and Y25130 together, BHSST-induced effects were completely blocked. Pretreatment with Ca 2+-free solution or thapsigargin inhibited BHSST-induced effects. Moreover, BHSST blocked both the transient receptor potential melastatin (TRPM) 7 and voltage-sensitive calcium-activated chloride (anoctamin-1, ANO1) channels. In normal mice, ITR% values were significantly increased by BHSST in a dose-dependent manner. The ITR% of GMD mice was significantly reduced relative to those of normal mice, which were significantly reversed by BHSST in a dose-dependent manner. Conclusion: These results suggested that BHSST depolarizes the pacemaker potentials of ICCs in a dose-dependent manner through the M 3 and 5-HT 3 receptors via internal and external Ca 2+-dependent and TRPM7-and ANO1independent pathways in vitro. Moreover, BHSST increased ITR% in vivo in normal mice and GMD mouse models. Taken together, the results of this study showed that BHSST had the potential for development as a prokinetic agent in GI motility function.

Interstitial Cells of Cajal: Potential Targets for Functional Dyspepsia Treatment Using Medicinal Natural Products

“…Regarding its underlying mechanisms, several preclinical studies have suggested an association with the recovery of ICCs. Banhasasim-tang depolarizes the slow-wave potentials in ICCs by regulating internal and external Ca2 + via the M 3 and 5-HT 3 receptors in a dose-dependent manner [44]. We also found that 4…”

“…ese results have been obtained mainly from cell-based experiments with ICCs using the patch-clamp technique. Many single-herb extracts [41,42], TCM drugs [14,26,[43][44][45][46][47][48][49][50][51], and herb-derived flavonoids [52] depolarize the pacemaker potential of ICCs in a dose-dependent manner. Depending on the herbal products, different receptors and their signaling pathways are involved in this depolarization process.…”

“…Molecular mechanism Depolarizers of the pacemaker potential of ICCs Gamisoyo-san [14], Citrus unshiu peel extracts [41], Dangkwisoo-san [43] Via M 3 receptor Magnolia officinalis [42] Via M 2 and M 3 receptors Banhasasim-tang [44] Via M 3 and 5-HT 3 receptors Daegeonjung-tang [45] Via M 3 and 5-HT 4 receptors Hesperidin, a citrus flavonoid [52] Via 5-HT 4 receptor Hwangryunhaedok-tang [46], Samhwangsasim-tang [47], Socheongryong-tang [48] Via 5-HT 3 and 5-HT 4 receptor Yukgunja-tang [49] Via ghrelin receptor Leejung-tang [90], Sengmek-san [50] Via PLC pathway Pyungwi-san [51] Via PKC pathway Inhibitors of the pacemaker potential of ICCs Galgeun-tang [26] Via α2-and β1-adrenoceptors Quercetin [91] Via opioid receptor signaling pathways M receptors: muscarinic acetylcholine receptors; 5-HT receptors: 5-hydroxytryptamine receptors; PLC: phospholipase C; PKC: protein kinase C. [53]. is decoction also promotes the numbers of stomach ICCs and the levels of SCF in diabetic rats [54].…”

“…Regarding its underlying mechanisms, several preclinical studies have suggested an association with the recovery of ICCs. Banhasasim-tang depolarizes the slow-wave potentials in ICCs by regulating internal and external Ca2 + via the M 3 and 5-HT 3 receptors in a dose-dependent manner [ 44 ]. We also found that Banhasasim-tang can reverse the inactivation of c-kit and modulate oxidative stress and contraction-related genes such as the 5-HT 4 receptor, ANO1, RYR receptor 3, and smooth muscle myosin light chain kinase in a loperamide-injected mouse model [ 53 ].…”

Network Pharmacological Analysis and Experimental Validation of the Effect of Smilacis Glabrae Rhixoma on Gastrointestinal Motility Disorder

“…Research on gastrointestinal motility abnormalities in traditional medicine has been actively conducted [ 31 ]. It was found that the possibility of controlling gastrointestinal movement and reduced gastrointestinal movement increased due to various traditional medicines, such as Pyungwi-san [ 32 ], Lizhong Tang [ 33 ], Poncirus trifoliate fruit [ 34 ], So-Cheong-Ryong-Tang [ 35 ], San-Huang-Xie-Xin-tang [ 36 ], Schisandra chinensis [ 37 ], Ge-Gen-Tang [ 38 ], Liriope Platyphylla Wang Et tang [ 39 ], Hwangryunhaedok-tang [ 40 ], Gamisoyo-San [ 41 ], Banhasasim-Tang [ 42 ], and Pinellia ternata Breitenbach [ 43 ]. In order for the oriental medical approach to become an efficient approach in controlling gastrointestinal motility, large-scale clinical studies are needed to prove the reproducibility and validity of the effects.…”