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 M2 receptor antagonist) did not inhibit BHSST-induced pacemaker potential depolarization. However, when we applied 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide (4-DAMP; a muscarinic M3 receptor antagonist), BHSST-induced effects were blocked. Pretreatment with Y25130 (a 5-HT3 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 Ca2+-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 M3 and 5-HT3 receptors via internal and external Ca2+-dependent and TRPM7- and ANO1-independent 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.

Journal Section:
Research Article
Keywords:
Banhasasim-tang, Interstitial cells of Cajal, Pacemaker potentials, Muscarinic receptor, 5-HT receptor, Intestinal transit rates, Intestinal motility
1.
Kawashima K, Nomura A, Makino T, Saito K, Kano Y. Pharmacological properties of traditional medicine (XXIX): effect of Hange-shashin-to and the combinations of its herbal constituents on rat experimental colitis. Biol Pharm Bull. 2004 Oct;27(10):1599–603.
2.
Kase Y, Saitoh K, Makino B, Hashimoto K, Ishige A, Komatsu Y. Relationship between the antidiarrhoeal effects of Hange-Shashin-To and its active components. Phytother Res. 1999 Sep;13(6):468–73.
3.
Fukamachi H, Matsumoto C, Omiya Y, Arimoto T, Morisaki H, Kataoka H, et al. Effects of hangeshashinto on growth of oral microorganisms. Evid Based Complement Alternat Med. 2015;2015:512947.
4.
Kono T, Kaneko A, Matsumoto C, Miyagi C, Ohbuchi K, Mizuhara Y, et al. Multitargeted effects of hangeshashinto for treatment of chemotherapy-induced oral mucositis on inducible prostaglandin E2 production in human oral keratinocytes. Integr Cancer Ther. 2014 Sep;13(5):435–45.
5.
Matsumoto C, Sekine-Suzuki E, Nyui M, Ueno M, Nakanishi I, Omiya Y, et al. Analysis of the antioxidative function of the radioprotective Japanese traditional (Kampo) medicine, hangeshashinto, in an aqueous phase. J Radiat Res (Tokyo). 2015 Jul;56(4):669–77.
6.
Hatakeyama H, Takahashi H, Oridate N, Kuramoto R, Fujiwara K, Homma A, et al. Hangeshashinto improves the completion rate of chemoradiotherapy and the nutritional status in patients with head and neck cancer. ORL J Otorhinolaryngol Relat Spec. 2015;77(2):100–8.
7.
Matsuda C, Munemoto Y, Mishima H, Nagata N, Oshiro M, Kataoka M, et al. Double-blind, placebo-controlled, randomized phase II study of TJ-14 (Hangeshashinto) for infusional fluorinated-pyrimidine-based colorectal cancer chemotherapy-induced oral mucositis. Cancer Chemother Pharmacol. 2015 Jul;76(1):97–103.
8.
Xu G. Treatment of reflux laryngopharyngitis with modified banxia xiexin tang (Pinellia decoction for draining the heart)—a report of 40 cases. J Tradit Chin Med. 2006 Jun;26(2):127–31.
9.
Lee KG, Cui X, Lim JP. Effect of the concurrent administration of Banhasasim-tang with cimetidine on gastric ulcer in rats. J Physiol & Pathol Korean Med. 2002;16:572–6.
10.
Kase Y, Hayakawa T, Takeda S, Ishige A, Aburada M, Okada M. Pharmacological studies on antidiarrheal effects of Hange-shashin-to. Biol Pharm Bull. 1996 Oct;19(10):1367–70.
11.
Takasuna K, Kasai Y, Kitano Y, Mori K, Kobayashi R, Hagiwara T, et al. Protective effects of kampo medicines and baicalin against intestinal toxicity of a new anticancer camptothecin derivative, irinotecan hydrochloride (CPT-11), in rats. Jpn J Cancer Res. 1995 Oct;86(10):978–84.
12.
Kase Y, Hayakawa T, Aburada M, Komatsu Y, Kamataki T. Preventive effects of Hange-shashin-to on irinotecan hydrochloride-caused diarrhea and its relevance to the colonic prostaglandin E2 and water absorption in the rat. Jpn J Pharmacol. 1997 Dec;75(4):407–13.
13.
Kawashima K, Fujimura Y, Makino T, Kano Y. Pharmacological properties of traditional medicine (XXXII): protective effects of hangeshashinto and the combinations of its major constituents on gastric lesions in rats. Biol Pharm Bull. 2006 Sep;29(9):1973–5.
14.
Huizinga JD, Thuneberg L, Klüppel M, Malysz J, Mikkelsen HB, Bernstein A. W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature. 1995 Jan;373(6512):347–9.
15.
Kim BJ, Lim HH, Yang DK, Jun JY, Chang IY, Park CS, et al. Melastatin-type transient receptor potential channel 7 is required for intestinal pacemaking activity. Gastroenterology. 2005 Nov;129(5):1504–17.
16.
Eisenman ST, Gibbons SJ, Verhulst PJ, Cipriani G, Saur D, Farrugia G. Tumor necrosis factor alpha derived from classically activated “M1” macrophages reduces interstitial cell of Cajal numbers. Neurogastroenterol Motil. 2017 Apr;29(4).
17.
He CL, Soffer EE, Ferris CD, Walsh RM, Szurszewski JH, Farrugia G. Loss of interstitial cells of cajal and inhibitory innervation in insulin-dependent diabetes. Gastroenterology. 2001 Aug;121(2):427–34.
18.
Zárate N, Mearin F, Wang XY, Hewlett B, Huizinga JD, Malagelada JR. Severe idiopathic gastroparesis due to neuronal and interstitial cells of Cajal degeneration: pathological findings and management. Gut. 2003 Jul;52(7):966–70.
19.
Hong NR, Park HS, Ahn TS, Kim HJ, Ha KT, Kim BJ. Ginsenoside Re inhibits pacemaker potentials via adenosine triphosphate-sensitive potassium channels and the cyclic guanosine monophosphate/nitric oxide-dependent pathway in cultured interstitial cells of Cajal from mouse small intestine. J Ginseng Res. 2015 Oct;39(4):314–21.
20.
Jun JY, Choi S, Chang IY, Yoon CK, Jeong HG, Kong ID, et al. Deoxycholic acid inhibits pacemaker currents by activating ATP-dependent K+ channels through prostaglandin E2 in interstitial cells of Cajal from the murine small intestine. Br J Pharmacol. 2005 Jan;144(2):242–51.
21.
Koh SD, Sanders KM, Ward SM. Spontaneous electrical rhythmicity in cultured interstitial cells of cajal from the murine small intestine. J Physiol. 1998 Nov;513(Pt 1):203–13.
22.
Tanila H, Kauppila T, Taira T. Inhibition of intestinal motility and reversal of postlaparotomy ileus by selective alpha 2-adrenergic drugs in the rat. Gastroenterology. 1993 Mar;104(3):819–24.
23.
Wichmann A, Allahyar A, Greiner TU, Plovier H, Lundén GÖ, Larsson T, et al. Microbial modulation of energy availability in the colon regulates intestinal transit. Cell Host Microbe. 2013 Nov;14(5):582–90.
24.
Wu YS, Lu HL, Huang X, Liu DH, Meng XM, Guo X, et al. Diabetes-induced loss of gastric ICC accompanied by up-regulation of natriuretic peptide signaling pathways in STZ-induced diabetic mice. Peptides. 2013 Feb;40:104–11.
25.
Kim HJ, Kim H, Jung MH, Kwon YK, Kim BJ. Berberine induces pacemaker potential inhibition via cGMP-dependent ATP-sensitive K+ channels by stimulating mu/delta opioid receptors in cultured interstitial cells of Cajal from mouse small intestine. Mol Med Rep. 2016 Oct;14(4):3985–91.
26.
Han S, Kim JS, Jung BK, Han SE, Nam JH, Kwon YK, et al. Effects of ginsenoside on pacemaker potentials of cultured interstitial cells of Cajal clusters from the small intestine of mice. Mol Cells. 2012 Mar;33(3):243–9.
27.
Huizinga JD, Chang G, Diamant NE, El-Sharkawy TY. Electrophysiological basis of excitation of canine colonic circular muscle by cholinergic agents and substance P. J Pharmacol Exp Ther. 1984 Dec;231(3):692–9.
28.
Inoue R, Chen S. Physiology of muscarinic receptor-operated nonselective cation channels in guinea-pig ileal smooth muscle. EXS. 1993;66:261–8.
29.
Gershon MD, Tack J. The serotonin signaling system: from basic understanding to drug development for functional GI disorders. Gastroenterology. 2007 Jan;132(1):397–414.
30.
Epperson A, Hatton WJ, Callaghan B, Doherty P, Walker RL, Sanders KM, Ward SM, Horowitz B. Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal. Am J Physiol Cell Physiol. 2000;279:C529–C539.
31.
Liu HN, Ohya S, Nishizawa Y, Sawamura K, Iino S, Syed MM, et al. Serotonin augments gut pacemaker activity via 5-HT3 receptors. PLoS One. 2011;6(9):e24928.
32.
Shahi PK, Choi S, Zuo DC, Yeum CH, Yoon PJ, Lee J, et al. 5-hydroxytryptamine generates tonic inward currents on pacemaker activity of interstitial cells of cajal from mouse small intestine. Korean J Physiol Pharmacol. 2011 Jun;15(3):129–35.
33.
Ward SM. Interstitial cells of Cajal in enteric neurotransmission. Gut. 2000 Dec;47(Suppl 4):iv40–3; discussion iv52.
34.
Zhu MH, Kim TW, Ro S, Yan W, Ward SM, Koh SD, et al. A Ca(2+)-activated Cl(-) conductance in interstitial cells of Cajal linked to slow wave currents and pacemaker activity. J Physiol. 2009 Oct;587(Pt 20):4905–18.
35.
Kim BJ, Lee GS, Kim HW. Involvement of transient receptor potential melastatin type 7 channels on Poncirus fructus-induced depolarizations of pacemaking activity in interstitial cells of Cajal from murine small intestine. Integr Med Res. 2013 Jun;2(2):62–9.
36.
Tan-No K, Niijima F, Nakagawasai O, Sato T, Satoh S, Tadano T. Development of tolerance to the inhibitory effect of loperamide on gastrointestinal transit in mice. Eur J Pharm Sci. 2003 Nov;20(3):357–63.
37.
Watanabe K, Matsuura K, Gao P, Hottenbacher L, Tokunaga H, Nishimura K, et al. Traditional Japanese Kampo medicine: clinical research between modernity and traditional medicine-the state of research and methodological suggestions for the future. Evid Based Complement Alternat Med. 2011;2011:513842.
38.
Bensky D, Barolet R. Chinese Herbal Medicine, Formulas & Strategies. Incorporated P.O. Box 12689, Seattle, Washington: Eastland press; 1990, pp. 150–2.
39.
Kase Y, Saitoh K, Ishige A, Komatsu Y. Mechanisms by which Hange-shashin-to reduces prostaglandin E2 levels. Biol Pharm Bull. 1998 Dec;21(12):1277–81.
40.
Kamide D, Yamashita T, Araki K, Tomifuji M, Shiotani A. Hangeshashinto (TJ-14) prevents radiation-induced mucositis by suppressing cyclooxygenase-2 expression and chemotaxis of inflammatory cells. Clin Transl Oncol. 2017 Nov;19(11):1329–36.
41.
Benham CD, Bolton TB, Lang RJ. Acetylcholine activates an inward current in single mammalian smooth muscle cells. Nature. 1985 Jul;316(6026):345–7.
42.
Inoue R, Isenberg G. Acetylcholine activates nonselective cation channels in guinea pig ileum through a G protein. Am J Physiol. 1990 Jun;258(6 Pt 1):C1173–8.
43.
Lee YM, Kim BJ, Kim HJ, Yang DK, Zhu MH, Lee KP, et al. TRPC5 as a candidate for the nonselective cation channel activated by muscarinic stimulation in murine stomach. Am J Physiol Gastrointest Liver Physiol. 2003 Apr;284(4):G604–16.
44.
Tsvilovskyy VV, Zholos AV, Aberle T, Philipp SE, Dietrich A, Zhu MX, et al. Deletion of TRPC4 and TRPC6 in mice impairs smooth muscle contraction and intestinal motility in vivo. Gastroenterology. 2009 Oct;137(4):1415–24.
45.
Lee KP, Jun JY, Chang IY, Suh SH, So I, Kim KW. TRPC4 is an essential component of the nonselective cation channel activated by muscarinic stimulation in mouse visceral smooth muscle cells. Mol Cells. 2005 Dec;20(3):435–41.
46.
Xia Y, Fu Z, Hu J, Huang C, Paudel O, Cai S, et al. TRPV4 channel contributes to serotonin-induced pulmonary vasoconstriction and the enhanced vascular reactivity in chronic hypoxic pulmonary hypertension. Am J Physiol Cell Physiol. 2013;305:C704–15.
47.
Chen H, Redelman D, Ro S, Ward SM, Ordög T, Sanders KM. Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine. Am J Physiol Cell Physiol. 2007 Jan;292(1):C497–507.
48.
So KY, Kim SH, Sohn HM, Choi SJ, Parajuli SP, Choi S, et al. Carbachol regulates pacemaker activities in cultured interstitial cells of Cajal from the mouse small intestine. Mol Cells. 2009 May;27(5):525–31.
49.
Shahi PK, Choi S, Zuo DC, Kim MY, Park CG, Kim YD, et al. The possible roles of hyperpolarization-activated cyclic nucleotide channels in regulating pacemaker activity in colonic interstitial cells of Cajal. J Gastroenterol. 2014 Jun;49(6):1001–10.
50.
Huang F, Rock JR, Harfe BD, Cheng T, Huang X, Jan YN, et al. Studies on expression and function of the TMEM16A calcium-activated chloride channel. Proc Natl Acad Sci USA. 2009 Dec;106(50):21413–8.
51.
Mikkelsen HB, Malysz J, Huizinga JD, Thuneberg L. Action potential generation, Kit receptor immunohistochemistry and morphology of steel-Dickie (Sl/Sld) mutant mouse small intestine. Neurogastroenterol Motil. 1998 Feb;10(1):11–26.
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