Depolarizing effects of daikenchuto on interstitial cells of cajal from mouse small intestine
Hyungwoo Kim1, Hyun Jung Kim2, Dongki Yang3, Myeong Ho Jung2, Byung Joo Kim2
1 Division of Pharmacology, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do 50612, Republic of Korea
2 Division of Longevity and Biofunctional Medicine and Healthy Aging Korean Medical Research Center, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do 50612, Republic of Korea
3 Department of Physiology, College of Medicine, Gachon University, Incheon, Republic of Korea
|Date of Submission||15-Dec-2015|
|Date of Acceptance||21-Jan-2016|
|Date of Web Publication||06-Jan-2017|
Dr. Byung Joo Kim
Division of Longevity and Biofunctional Medicine,Pusan National University School of KoreanMedicine, 49 Busandaehakro, Mulgeum-eup,Yangsan, Gyeongsangnam-do 50612
Republic of Korea
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Daikenchuto (DKT; TJ-100, TU-100)a traditional herbal medicineis used in modern medicine to treat gastrointestinal (GI) functional disorders. Interstitial cells of Cajal (ICCs) are the pacemaker cells of the GI tract and play important roles in the regulation of GI motility. Objective: The objective of this study was to investigate the effects of DKT on the pacemaker potentials (PPs) of cultured ICCs from murine small intestine. Materials and Methods: Enzymatic digestions were used to dissociate ICCs from mouse small intestine tissues. All experiments on ICCs were performed after 12 h of culture. The whole-cell patch-clamp configuration was used to record ICC PPs (current clamp mode). All experiments were performed at 30-32°C. Results: In current-clamp modeDKT depolarized and concentration-dependently decreased the amplitudes of PPs. Y25130 (a 5-HT3 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist) did not block DKT-induced PP depolarizationbut RS39604 (a 5-HT4 receptor antagonist) did. Methoctramine (a muscarinic M2 receptor antagonist) failed to block DKT-induced PP depolarizationbut pretreating 4-diphenylacetoxy-N-methylpiperidine methiodide (a muscarinic M3 receptor antagonist) facilitated blockade of DKT-induced PP depolarization. Pretreatment with an external Ca2+-free solution or thapsigargin abolished PPsand under these conditionsDKT did not induce PP depolarization. FurthermoreGinseng radix and Zingiberis rhizomes depolarized PPswhereas Zanthoxyli fructus fruit (the third component of DKT) hyperpolarized PPs. Conclusion: These results suggest that DKT depolarizes ICC PPs in an internal or external Ca2+-dependent manner by stimulating 5-HT4 and M3 receptors. Furthermorethe authors suspect that the component in DKT largely responsible for depolarization is probably also a component of Ginseng radix and Zingiberis rhizomes.
Abbreviation used: DKT: DaikenchutoGI: GastrointestinalICCs: Interstitial cells of CajalPPs: Pacemaker Potentials.
Keywords: Daikenchuto, gastrointestinal tract, interstitial cells of Cajal, pacemaker potentials
|How to cite this article:|
Kim H, Kim HJ, Yang D, Jung MH, Kim BJ. Depolarizing effects of daikenchuto on interstitial cells of cajal from mouse small intestine. Phcog Mag 2017;13:141-7
|How to cite this URL:|
Kim H, Kim HJ, Yang D, Jung MH, Kim BJ. Depolarizing effects of daikenchuto on interstitial cells of cajal from mouse small intestine. Phcog Mag [serial online] 2017 [cited 2021 Nov 30];13:141-7. Available from: http://www.phcog.com/text.asp?2017/13/49/141/196312
- Daikenchuto (DKT) depolarized and concentration-dependently decreased the amplitudes of pacemaker potentials (PPs)
- Y25130 (a 5-HT3 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist) did not block DKT-induced PP depolarizationbut RS39604 (a 5-HT4 receptor antagonist) did
- Methoctramine (a muscarinic M2 receptor antagonist) failed to block DKT-induced PP depolarizationbut pretreating 4-DAMP (a muscarinic M3 receptor antagonist) facilitated blockade of DKT-induced PP depolarization
- Ginseng radix and Zingiberis rhizomes depolarized PPswhereas Zanthoxyli fructus fruit (the third component of DKT) hyperpolarized PPs.
| Introduction|| |
Chinese (or Japanese) traditional herbal medicines are prescribed for the treatment of a wide array of diseases and conditionsincluding gastrointestinal (GI) disorders. Daikenchuto (DKT; TJ-100, TU-100) is the most widely prescribed traditional Chinese herbal medicineand it is called Kampo in Japan. DKT is a mixture of Ginseng radix, Zingiberis siccatum rhizomes, and Zanthoxyli fructus, is used traditionally to treat abdominal bloating and a cold sensation in the abdomen,,, and prescribed by physicians for the treatment of chronic constipation.
Interstitial cells of Cajal (ICCs) are the pacemaker cells of the GI tract and generate rhythmic oscillations in membrane potentials known as slow waves,, and thusICCs play important roles in the regulation of GI motility. Endogenous agentssuch as neurotransmitters, hormones, and paracrine substances, modulate GI tract motility by influencing ICCs,,, and it appears that some traditional herbal medicines could regulate the pacemaker potentials (PPs) of ICCs.,, It has been reported that DKT accelerates small intestinal movement by directly inhibiting smooth muscle and partially by inhibiting neural activities, and that it induces phasic contractions in the duodenum and proximal jejunum via cholinergic receptors. These findings provide a scientific basis suggesting the therapeutic use of DKT for the treatment of GI disorders. Howeverrelatively little is known about the effects of DKT on the PPs of ICCs in the GI tract. Thereforein the present studywe investigated the effects of DKT on the PPs of cultured ICCs obtained from mouse small intestine.
| Materials and Methods|| |
Preparation of daikenchuto and its constituents
DKT extract powder was manufactured by Tsumura and Co.Tokyo. To prepare DKT constituentsthe powders obtained from the ethanol or water extracts of Ginseng radix (Catalog number: CA03-041), Zingiberis rhizome (Catalog number: CA04-001), and Zanthoxyli fructus (Catalog number: 029-037) were obtained from the plant extract bank at the Korean Research Institute of Bioscience and Biotechnology (Daejeon, Korea). DKT and its constituents were authenticated by Hyungwoo Kim (Division of Pharmacology, Pusan National University, School of Korean Medicine, Yangsan, Korea). The powder was then immersed in ethanol or water, sonicated for 15 min, and extracted for 72 h. The extract so obtained was filtered through nonfluorescent cotton and evaporated under reduced pressure using a rotary evaporator (N-1000 SWD, Eyela, Japan) in 45°C. The condensed extract was then lyophilized using a Modul Spin 40 dryer (Biotron Corporation, Calgary, Canada) for 24 h. The yield of lyophilized powder obtained was 12.3%. The DKT was dissolved in distilled water at a concentration of 0.5 g (crude drug)/ml and stored in a refrigerator. The extracts of Ginseng radix, Zingiberis rhizome, and Zanthoxyli fructus were dissolved in dimethyl sulfoxide as a stock solution at 100 mg/mL and stored at 4°C. The stock solution was diluted with medium to the desired concentration prior to use.
Preparation of cells and cell cultures
Animal care and the study protocol were in accordance with the guidelines issued by the Ethics Committee of Pusan National University (Yangsan, Republic of Korea). BALB/c mice (3-5 day-old) were used throughout the study. Small intestines were excised (from 1 cm below the pyloric ring to the cecum) and opened along the mesenteric border. Luminal contents were removed using Krebs-Ringer bicarbonate solution, and the tissues were pinned to the bases of Sylgard dishes. Mucosae were removed by sharp dissection. Small tissue strips of intestine muscle (consisting of circular and longitudinal muscles) were equilibrated for 30 min in Ca2+-free Hank's solutionwhich contained the following; potassium chloride (KCl), 5.36 mM; sodium chloride (NaCl), 125 mM; sodium hydroxide (NaOH), 0.34 mM; sodium bicarbonate, 0.44 mM; glucose, 10 mM; sucrose, 2.9 mM; and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 11 mM (final pH of 7.4). Cells were then dispersed in an enzyme solution containing collagenase (Worthington Biochemical, Lakewood, NJ, USA; 1.3 mg/mL)bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA; 2 mg/mL), trypsin inhibitor (Sigma-Aldrich; 2 mg/mL), and ATP (0.27 mg/mL), and plated onto sterile glass coverslips coated with murine collagen (2.5 mg/mL; Falcon/BD, Franklin Lakes, NJ, USA) in 35 mm culture dishes. Cells were cultured at 37°C in a 95% oxygen/5% carbon dioxide incubator in smooth muscle growth medium (Clonetics, San Diego, CA, USA) supplemented with 2% antibiotics/antimycotics (Gibco, Grand Island, NY, USA) and murine stem cell factor (5 ng/mL; Sigma-Aldrich). All experiments on ICC clusters were performed after they had been cultured for 12 h. ICCs were identified immunologically using an anti-c-Kit antibody, that is, phycoerythrin-conjugated rat anti-mouse c-Kit monoclonal antibody (eBioscience, San Diego, CA, USA), at a dilution of 1:50 for 20 min. Because the morphology of ICCs differed from other cell types in culture, it could be identified under a phase-contrast microscope after incubation with anti-c-Kit antibody.
Physiological salt solution was used to bathe cultured ICC clusters (Na+-Tyrode) and contained the following: KCl5 mM; NaCl, 135 mM; calcium chloride (CaCl2), 2 mM; glucose10 mM; magnesium chloride (MgCl2), 1.2 mM; and HEPES, 10 mM (adjusted to pH 7.4 with NaOH). The pipette solution used to examine pacemaker activity contained the following: KCl, 140 mM; MgCl2, 5 mM; dipotassium ATP (K2ATP), 2.7 mM; sodium GTP (NaGTP), 0.1 mM; creatine phosphate disodium, 2.5 mM; HEPES, 5 mM; and ethylene glycol tetra-acetic acid, 0.1 mM (adjusted to pH 7.2 with potassium hydroxide). Patch-clamp techniques were conducted in whole-cell configuration to record potentials (i.e.current clamp mode) from cultured ICCs using Axopatch I-D and Axopatch 200B amplifiers (Axon Instruments, Foster, CA, USA). Command pulses were applied using an IBM-compatible personal computer (Compaq; Houston, TX, USA) and pClamp software (versions 6.1 and 10.0; Axon InstrumentsFoster City, CA, USA). Data were filtered at 5 kHz and displayed on an oscilloscopea computer monitorand/or a pen recorder (Gould 2200; Gould, Valley View, OH, USA). Results were analyzed using pClamp and Origin software (version 6.0, Microcal, Northampton, MA, USA). All experiments were performed at 30-33°C.
The drugs used in the experiments were Y25130, RS39604, SB269970, methoctramine, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), and thapsigargin, and all were purchased from Sigma-Aldrich (St. Louis, MO, USA). Stock solutions were prepared and stored according to the manufacturer's instructions. Chemicals were dissolved in physiological salt solution to their final concentrations immediately before use.
Results are expressed as mean ± standard errors. The Student's t-test was used to determine the significances of differences. A statistically significant difference was seen at P < 0.05. "n" values reported in the text refer to the number of cells used in patch-clamp experiments.
| Results|| |
Effects of daikenchuto on the pacemaker potentials of cultured interstitial cells of Cajal from murine small intestine
The patch-clamp technique was applied to ICCs that formed network-like structures after culture for 12 h. Under current clamp mode (I = 0)ICCs generated PPs [Figure 1]a with a mean resting membrane potential of -51.3 ± 2.3 mV and a mean amplitude of 25.4 ± 1.5 mV. DKT (1-10 mg/ml) depolarized PPs and decreased the PP amplitudes in a concentration-dependent manner [Figure 1]b-d. In the presence of DKT mean degrees of depolarization were 2.6 ± 0.6 mV at 1 mg/ml, 16.2 ± 0.7 mV at 5 mg/ml, and 26.1 ± 1.5 mV at 10 mg/ml [[Figure 1]e, n = 24] and mean amplitudes were 24.3 ± 1.3 mV at 1 mg/ml, 11.4 ± 1.4 mV at 5 mg/ml, and 7.2 ± 0.8 mV at 10 mg/ml [[Figure 1]f, n = 24]. Summarized values and a bar graph showing the effects of DKT on PPs are provided in [Figure 1]e and f. These results suggest that DKT dose dependently depolarizes ICC PPs.
|Figure 1: Effects of daikenchuto on pacemaker potentials in cultured interstitial cells of Cajal from mouse small intestine. (a-d) The pacemaker potentials of interstitial cells of Cajal exposed to daikenchuto (1-10 mg/ml) in current clamp mode (i = 0). Responses to daikenchuto (e and f). Bars represent mean ± standard error of the means. *P < 0.05**P < 0.01. Significantly different from nontreated controls. CTRL: Control|
Click here to view
Identification of daikenchuto receptor subtypes in cultured interstitial cells of Cajal from murine small intestine
To investigate the relationship between DKT and its receptors, we studied 5-HT and muscarinic receptors because they are known to mediate GI tract motility and to be strongly associated with prokinetic activity.,, The stimulation of 5-HT4 receptor (5-HT4R) in the enteric nervous system results in the release of acetylcholine in the GI tract, which leads to the excitation of smooth muscles in myenteric plexusand thus, 5-HT4R is regarded a prokinetic. Thereforewe investigated whether the prokinetic action of DKT involves 5-HT receptors. In ICCsonly three receptors (5-HT3R5-HT4Rand 5-HT7R) are present.,, To identify the 5-HT receptor subtypes involved in the effects of DKTICCs were pretreated with various 5-HT receptor antagonists and then treated with DKT. Y25130 (a 5-HT3 receptor antagonist), RS39604 (a 5-HT4R antagonist), and SB269970 (a 5-HT7 receptor antagonist) were all pretreated at 10 μM for 5 min, and then DKT was added. After pretreating Y25130 or SB269970DKT depolarized membranes [Figure 2]a and d; membrane depolarization produced in the presence of Y25130 or SB269970 by DKT (5 mg/ml) was 15.8 ± 1.0 mV and 16.5 ± 0.6 mV, respectively [n = 4; [Figure 2]d]. However, pretreatment with RS39604 blocked the effect of DKT [n = 4; [Figure 2]b and d]. In addition, ICCs isolated from the GI tract express M2 and M3 subtypes of muscarinic receptors. To identify the muscarinic receptor subtypes involved, ICCs were pretreated with muscarinic receptor antagonists and then treated with DKT. Methoctramine (a muscarinic M2 receptor antagonist) or 4-DAMP (a muscarinic M3 receptor antagonist) were pretreated at 10 μM for 5 min and then DKT (10 mg/ml) was added. Treatment with methoctramine or 4-DAMP had no effect on PPsand pretreatment with methoctramine did not block the DKT-induced PP depolarization [Figure 3]a. In the presence of methoctramine, mean DKT-induced PP depolarization was 26.3 ± 0.5 mV [n = 4; [Figure 3]c], but after pretreating 4-DAMP, DKT-induced PP depolarization was blocked [Figure 3]b. After 4-DAMP pretreatment, DKT-induced PPs depolarization was 0.3 ± 0.4 mV [n = 4; [Figure 3]c]. These results show that DKT affects ICCs through 5-HT4 and M3 receptors.
|Figure 3: Effects of muscarinic receptor subtype antagonists on daikenchuto-induced pacemaker potential depolarization in cultured interstitial cells of Cajal from mouse small intestine. Pacemaker potentials were depolarized when interstitial cells of Cajal were exposed to daikenchuto (10 mg/ml) in the presence of methoctramine (a muscarinic M2 receptor antagonist; 10 μM) (a)in the presence of 4-DAMP (a muscarinic M3 receptor antagonist; 10 μM) (b). Responses to daikenchuto in the presence of different receptor antagonists (c). Bars represent mean ± standard error of the means. **P < 0.01. Significantly different from non-treated controls. CTRL: ControlMethoct: Methoctramine|
Click here to view
Effects of external Ca2+-free solution and of Ca2+-ATPase inhibitor on daikenchuto-induced pacemaker potential depolarization in cultured interstitial cells of Cajal from murine small intestine
External and internal Ca2+ regulation plays important roles in smooth muscle contraction and in the pacemaker activities of ICCs in the GI tract. To investigate the roles of external or internal Ca2+, DKT was applied under external Ca2+-free conditions and in the presence of thapsigargin (an inhibitor of Ca2+-ATPase in endoplasmic reticulum)., Pretreatment with external Ca2+-free solution abolished PPsand under this conditionDKT did not induce PP depolarization [n = 5; [Figure 4]a]. In addition, pretreatment with thapsigargin abolished PPsand similarly under this condition, DKT did not induce PP depolarization [n = 5; [Figure 4]b]. Summarized values and a bar graph showing the effects of DKT using an external Ca2+-free solution or in the presence of a Ca2+-ATPase inhibitor are shown in [Figure 4]c. These results show that DKT-induced PP depolarization is dependent on internal and external Ca2+ regulation.
|Figure 4: Effects of an external Ca2+-free solution and of thapsigargin on daikenchuto-induced pacemaker potential depolarization in cultured interstitial cells of Cajal from mouse small intestine. (a) External Ca2+-free solution and (b) thapsigargin (5 μM) abolished the generation of pacemaker potentials and blocked daikenchuto-induced pacemaker potential depolarization. (c) Responses to daikenchuto in external Ca2+-free solution and in the presence of thapsigargin are summarized. Bars represent mean ± standard error of the means. **P < 0.01. Significantly different from nontreated controls. CTRL: ControlThapsi: Thapsigargin|
Click here to view
Effects of daikenchuto constituents on the pacemaker potentials of cultured interstitial cells of Cajal from murine small intestine
We examined the effects of the constituents of DKTthat isGinseng radixZingiberis (ginger) rhizomesand Zanthoxyli fructus on the PPs of ICCs. Both Ginseng and dried ginger depolarized and decreased the amplitudes of PPs [Figure 5]a and b. However, Zanthoxylum fruit inhibited, hyperpolarized, and decreased the amplitudes of PPs [Figure 5]c. These results suggest that the main depolarizing component in DKT is probably also an ingredient of Ginseng radix and Zingiberis rhizomes.
| Discussion|| |
In the present studywe investigated the effect of DKT on GI motility by examining the PPs of ICCs from murine small intestine. In these cellsDKT depolarized PPs in an internal and an external Ca2+-dependent manner by stimulating 5-HT4 and M3 receptorswhich suggests that DKT offers a basis for developing novel prokinetic agents that prevent or alleviate GI motility dysfunctions. Furthermore, the study also shows that the primary depolarizing component in DKT is probably also present in Ginseng radix and in Zingiberis rhizomes.
Herbal therapy has been used in Asia for thousands of years. DKT is composed of Ginseng radix, Zingiberis siccatum rhizome, and Zanthoxyli fructus,  and it is often used to treat GI hypomotility, such as, ileus, following abdominal surgery. It has been shown that DKT induces contractions in the antrum, duodenum, and jejunum by acting through cholinergic and 5-HT3 receptors., On the other hand, Zanthoxylum fruit elicited contractions mainly in duodenum and jejunum, whereas dried ginger rhizome induced contractions in the antrum only and Ginseng root had no effect., Furthermore, the effects of DKT have been reported to be dependent on anatomic site and timing of its administration. During the fasting state, DKT had evident prokinetic effects. In additionin isolated guinea pig ileum, DKT was found to elicit contractile responses via muscarinic and 5-HT4Rs and to ameliorate morphine-induced GI transit disorder in mice as determined by colon transit times. In one study, it was suggested that small intestine motility improvement by DKT was due to its ginger root and Ginseng components via smooth muscle and neural inhibition. In addition, Kito et al. suggested that DKT had no effect on pacemaker mechanisms and electrical coupling between ICCs and smooth muscle cells in mouse small intestineand that thereforeit may contract smooth muscles by depolarizing membranes directly.
Most 5-HT are found in the GI tractand abnormalities in 5-HT signaling or metabolism are associated with several GI tract disorders, such as, dyspepsia, nausea, vomiting, coeliac disease, inflammatory bowel disease, and irritable bowel syndrome (IBS).,, 5-HT4R is expressed in several different cell types in intestinewhere it stimulates intestinal activity and is a target for the treatment of constipation-predominant IBS and chronic constipation., 5-HT7 receptors are often located near 5-HT4Rs where they augment 5-HT induced responses., Furthermore, 5-HT3 receptor antagonists are known to decrease colonic motility, secretion, and nociception, and are currently targeted to treat diarrhea-predominant IBS., Acetylcholine is involved in the control of almost all the functions of GI organ systems, and muscarinic receptors, which are commonly expressed in the GI tractare important for organ function., The tissues and cell types that express receptors are numerous and include ICCs and smooth muscle and mucosal cells in the stomach and intestine. Muscarinic receptors are classified into five subtypes, that ismuscarinic M1, M2, M3, M4, and M5 receptors,, and belong to the family of G protein-coupled receptors - heterotrimeric guanine nucleotide-binding proteins that regulate second messengers and ion channels. Three 5-HT receptors (5-HT35-HT4and 5-HT7) and two muscarinic receptors (M2 and M3) are present on ICCs.,,, In the present study, pretreatment with Y25130 (a 5-HT3 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist) facilitated DKT-induced membrane depolarization [Figure 2]a and c, but pretreatment with RS39604 (a 5-HT4R antagonist) blocked the effect of DKT [Figure 2]b. On the other hand, methoctramine (a muscarinic M2 receptor antagonist) did not block DKT-induced PP depolarization [Figure 3]a but pretreatment with 4-DAMP (a muscarinic M3 receptor antagonist) blocked DKT-induced PP depolarization [Figure 3]b. In addition, in GI tractgenerally both M2 and M3 were involved in GI motility. HoweverGI tract is composed of smooth muscleenteric nervous system and ICCs, and so on. Therefore, we think that DKT may act on only M3 receptor in ICCs. In addition, So et al. suggested that the modulation of pacemaker currents by carbachol in ICCs is mediated by only muscarinic M3 receptors not M2 receptors. Therefore, we think that DKT could modulate the PPs through only muscarinic M3 receptors in ICCs such as carbachol. These findings indicate that DKT affects ICCs through 5-HT4 and M3 receptors and that these receptors have important roles in the modulation of GI motility.
ICCs are the pacemaker cells of the GI tract and generate and propagate the slow waves that regulate GI motility, and networks of ICCs are never static even during physiological conditions. Apoptosis and transdifferentiation cause loss of ICCsand these losses can be restored by the proliferation and differentiation of stem cells or ameliorated by increasing the survival of ICCs., Losses of or deficiencies in ICCs have been observed in intestines of animal models of GI dysfunction and are believed to contribute to the development of motility disorders., ThereforeICCs play a critical physiological role in the coordination of intestinal contractile activity and constitute an important aspect of intestinal motility.
DKT is composed of Ginseng radix, Zingiberis siccatum rhizomeand Zanthoxyli fructus, and they have influences on the regulation of GI tract motility. Ginseng radix affects the GI tract. Ginseng increases mouse intestinal movement and promotes the relaxation of circular muscles in the gastric body. In isolated guinea pig GI tract tissues, Ginseng increases longitudinal muscle contraction in the ileum and distal colon and in the rabbit intestine, Ginseng stimulates intestinal motility. Furthermore, ginsenoside Re regulates the pacemaking activity of ICCs in mouse small intestine. Zingiberis siccatum rhizome continuously decreased the amplitude of contraction and Zanthoxyli fructus increased jejunal contraction in isolated rabbit jejunum.
In the present studyDKT depolarized PPs in ICCsand Ginseng and dried ginger also had depolarizing effects [Figure 5]a and b, whereas Zanthoxylum fruit hyperpolarized PPs [Figure 5]c, which suggests that the main depolarizing component in DKT is probably a common component of Ginseng radix and Zingiberis rhizome.
| Conclusion|| |
The present study shows that in ICCsDKT depolarizes and decreases the amplitudes of PPs in a concentration-dependent manner via internal or external Ca2+. RS39604 (a 5-HT4R antagonist) and 4-DAMP (a muscarinic M3 receptor antagonist) blocked DKT-induced PP depolarization. Ginseng radix and Zingiberis rhizome depolarized ICC PPswhereas Zanthoxyli fructus fruit had a hyperpolarizing effect. We suggest that the main depolarizing component in DKT is probably an ingredient in Ginseng radix and Zingiberis rhizomes. These findings suggest that DKT is a good candidate for the development of a prokinetic agent.
This study was supported by The Korean National Research Foundation (NRF) funded by the Korean government (MSIP) (Grant no. 2014R1A5A2009936).
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Kono T, Shimada M, Yamamoto M, Kaneko A, Oomiya Y, Kubota K, et al
. Complementary and synergistic therapeutic effects of compounds found in Kampo medicine: Analysis of daikenchuto. Front Pharmacol 2015;6:159-
Kono T, Mishima H, Shimada M, Morita S, Sakamoto J, GONE Investigators. Preventive effect of goshajinkigan on peripheral neurotoxicity of FOLFOX therapy: A placebo-controlled double-blind randomized phase II study (the GONE Study). Jpn J Clin Oncol 2009;39:847-9.
Manabe N, Camilleri M, Rao A, Wong BS, Burton D, Busciglio I, et al
. Effect of daikenchuto (TU-100) on gastrointestinal and colonic transit in humans. Am J Physiol Gastrointest Liver Physiol 2010;298:G970-5.
Iturrino J, Camilleri M, Wong BS, Linker Nord SJ, Burton D, Zinsmeister AR, Randomised clinical trial: The effects of daikenchuto, TU-100, on gastrointestinal and colonic transit, anorectal and bowel function in female patients with functional constipation. Aliment Pharmacol Ther 2013;37:776-85.
Shimada M, Morine Y, Nagano H, Hatano E, Kaiho T, Miyazaki M, et al
. Effect of TU-100, a traditional Japanese medicine, administered after hepatic resection in patients with liver cancer: A multi-center, phase III trial (JFMC40-1001). Int J Clin Oncol 2015;20:95-104.
Iizuka N, Hamamoto Y, Constipation and herbal medicine. Front Pharmacol 2015;6:73-
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;373:347-9.
Sanders KM, A case for interstitial cells of Cajal as pacemakers and mediators of neurotransmission in the gastrointestinal tract. Gastroenterology 1996;111:492-515.
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;129:1504-17.
Lee JH, Kim SY, Kwon YK, Kim BJ, So I, Characteristics of the cholecystokinin-induced depolarization of pacemaking activity in cultured interstitial cells of Cajal from murine small intestine. Cell Physiol Biochem 2013;31:542-54.
Kim BJ, Kim SY, Lee S, Jeon JH, Matsui H, Kwon YK, et al
. The role of transient receptor potential channel blockers in human gastric cancer cell viability. Can J Physiol Pharmacol 2012;90:175-86.
Nam JH, Kim WK, Kim BJ, Sphingosine and FTY720 modulate pacemaking activity in interstitial cells of Cajal from mouse small intestine. Mol Cells 2013;36:235-44.
Kim BJ, Kim HW, Lee GS, Choi S, Jun JY, So I, et al
. Poncirus trifoliate fruit modulates pacemaker activity in interstitial cells of Cajal from the murine small intestine. J Ethnopharmacol 2013;149:668-75.
Hwang MW, Kim JN, Song HJ, Lim B, Kwon YK, Kim BJ, Effects of Lizhong Tang on cultured mouse small intestine interstitial cells of Cajal. World J Gastroenterol 2013;19:2249-55.
Lee S, Gim H, Shim JH, Jung Kim H, Lee JR, Kim SC, et al
. The traditional herbal medicine, Ge-Gen-Tang, inhibits pacemaker potentials by nitric oxide/cGMP dependent ATP-sensitive K(+) channels in cultured interstitial cells of Cajal from mouse small intestine. J Ethnopharmacol 2015;170:201-9.
Satoh K, Kase Y, Hayakawa T, Murata P, Ishige A, Sasaki H, Dai-kenchu-to enhances accelerated small intestinal movement. Biol Pharm Bull 2001;24:1122-6.
Jin XL, Shibata C, Naito H, Ueno T, Funayama Y, Fukushima K, et al
. Intraduodenal and intrajejunal administration of the herbal medicine, dai-kenchu-tou, stimulates small intestinal motility via cholinergic receptors in conscious dogs. Dig Dis Sci 2001;46:1171-6.
Gershon MD, Tack J, The serotonin signaling system: From basic understanding to drug development for functional GI disorders. Gastroenterology 2007;132:397-414.
Cavalcante Morais T, Cavalcante Lopes S, Bezerra Carvalho KM, Rodrigues Arruda B, Correia de Souza FT, Salles Trevisan MT, et al
. Mangiferin, a natural xanthone, accelerates gastrointestinal transit in mice involving cholinergic mechanism. World J Gastroenterol 2012;18:3207-14.
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;15:129-35.
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:e24928-
Epperson A, Hatton WJ, Callaghan B, Doherty P, Walker RL, Sanders KM, et al
. Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal. Am J Physiol Cell Physiol 2000;279:C529-39.
Ward SM, Ordog T, Koh SD, Baker SA, Jun JY, Amberg G, et al
. Pacemaking in interstitial cells of Cajal depends upon calcium handling by endoplasmic reticulum and mitochondria. J Physiol 2000;525:355-61.
Koh SD, Jun JY, Kim TW, Sanders KM, A Ca2+
-inhibited non-selective cation conductance contributes to pacemaker currents in mouse interstitial cell of Cajal. J Physiol 2002;540:803-14.
Sung TS, Kim HU, Kim JH, Lu H, Sanders KM, Koh SD, Protease-activated receptors modulate excitability of murine colonic smooth muscles by differential effects on interstitial cells. J Physiol 2015;593:1169-81.
Itoh T, Yamakawa J, Mai M, Yamaguchi N, Kanda T, The effect of the herbal medicine Dai-Kenchu-to on post-operative ileus. J Int Med Res 2002;30:428-32.
Suzuki H, Inadomi JM, Hibi T, Japanese herbal medicine in functional gastrointestinal disorders. Neurogastroenterol Motil 2009;21:688-96.
Suzuki H, Nishizawa T, Hibi T, Therapeutic strategies for functional dyspepsia and the introduction of the Rome III classification. J Gastroenterol 2006;41:513-23.
Shibata C, Sasaki I, Naito H, Ueno T, Matsuno S, The herbal medicine Dai-Kenchu-Tou stimulates upper gut motility through cholinergic and 5-hydroxytryptamine 3 receptors in conscious dogs. Surgery 1999;126:918-24.
Kawasaki N, Nakada K, Nakayoshi T, Furukawa Y, Suzuki Y, Hanyu N, et al
. Effect of Dai-kenchu-to on gastrointestinal motility based on differences in the site and timing of administration. Dig Dis Sci 2007;52:2684-94.
Satoh K, Hayakawa T, Kase Y, Ishige A, Sasaki H, Nishikawa S, et al
. Mechanisms for contractile effect of Dai-kenchu-to in isolated guinea pig ileum. Dig Dis Sci 2001;46:250-6.
Nakamura T, Sakai A, Isogami I, Noda K, Ueno K, Yano S, Abatement of morphine-induced slowing in gastrointestinal transit by Dai-kenchu-to, a traditional Japanese herbal medicine. Jpn J Pharmacol 2002;88:217-21.
Tack J, Talley NJ, Camilleri M, Holtmann G, Hu P, Malagelada JR, et al
. Functional gastroduodenal disorders. Gastroenterology 2006;130:1466-79.
Kito Y, Suzuki H, Effects of Dai-kenchu-to on spontaneous activity in the mouse small intestine. J Smooth Muscle Res 2006;42:189-201.
Beattie DT, Smith JA, Serotonin pharmacology in the gastrointestinal tract: A review. Naunyn Schmiedebergs Arch Pharmacol 2008;377:181-203.
De Ponti F, Drug development for the irritable bowel syndrome: Current challenges and future perspectives. Front Pharmacol 2013;4:7-
Spiller R, Serotonin and GI clinical disorders. Neuropharmacology 2008;55:1072-80.
Irving HR, Tan YY, Tochon-Danguy N, Liu H, Chetty N, Desmond PV, et al
. Comparison of 5-HT4 and 5-HT7 receptor expression and function in the circular muscle of the human colon. Life Sci 2007;80:1198-205.
Cellek S, Thangiah R, Jarvie EM, Vivekanandan S, Lalude O, Sanger GJ, Synergy between 5-HT4 receptor activation and acetylcholinesterase inhibition in human colon and rat forestomach. Neurogastroenterol Motil 2008;20:539-45.
Prins NH, Briejer MR, Van Bergen PJ, Akkermans LM, Schuurkes JA, Evidence for 5-HT7 receptors mediating relaxation of human colonic circular smooth muscle. Br J Pharmacol 1999;128:849-52.
Uchiyama T, Chess-Williams R, Muscarinic receptor subtypes of the bladder and gastrointestinal tract. J Smooth Muscle Res 2004;40:237-47.
Lecci A, Santicioli P, Maggi CA, Pharmacology of transmission to gastrointestinal muscle. Curr Opin Pharmacol 2002;2:630-41.
Caulfield MP, Birdsall NJ, International union of pharmacology. XVII. Classification of muscarinic acetylcholine receptors. Pharmacol Rev 1998;50:279-90.
Eglen RM, Muscarinic receptor subtypes in neuronal and non-neuronal cholinergic function. Auton Autacoid Pharmacol 2006;26:219-33.
Lanzafame AA, Christopoulos A, Mitchelson F, Cellular signaling mechanisms for muscarinic acetylcholine receptors. Receptors Channels 2003;9:241-60.
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;27:525-31.
Farrugia G, Interstitial cells of Cajal in health and disease. Neurogastroenterol Motil 2008;20:54-63.
Gibbons SJ, De Giorgio R, Faussone Pellegrini MS, Garrity-Park MM, Miller SM, Schmalz PF, et al
. Apoptotic cell death of human interstitial cells of Cajal. Neurogastroenterol Motil 2009;21:85-93.
Ordög T, Takayama I, Cheung WK, Ward SM, Sanders KM, Remodeling of networks of interstitial cells of Cajal in a murine model of diabetic gastroparesis. Diabetes 2000;49:1731-9.
Grover M, Bernard CE, Pasricha PJ, Lurken MS, Faussone-Pellegrini MS, Smyrk TC, et al
. Clinical-histological associations in gastroparesis: Results from the Gastroparesis Clinical Research Consortium. Neurogastroenterol Motil 2012;24:531-9.e249-
Furukawa Y, Shiga Y, Hanyu N, Hashimoto Y, Mukai H, Nishikawa K, et al
. Effect of Chinese herbal medicine on gastrointestinal motility and bowel obstruction. Jpn J Gastroenterol Surg 1995;28:956-60.
Hashimoto K, Satoh K, Kase Y, Ishige A, Kubo M, Sasaki H, et al
. Modulatory effect of aliphatic acid amides from Zanthoxylum piperitum
on isolated gastrointestinal tract. Planta Med 2001;67:179-81.
Murata P, Hayakawa T, Satoh K, Kase Y, Ishige A, Sasaki H, Effects of Dai-kenchu-to, a herbal medicine, on uterine and intestinal motility. Phytother Res 2001;15:302-6.
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;39:314-21.
Hayakawa T, Kase Y, Saito K, Hashimoto K, Ishige A, Komatsu Y, et al
. Pharmacological studies of the effect of Dai-kenchu-to on spontaneous contraction of isolated rabbit jejunum. J Smooth Muscle Res 1999;35:55-62.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]