J Physiol & Pathol Korean Med 28(6):630~635, 2014 생쥐소장카할세포의내향성향도잡이전압에미치는 내소산의억제효과에관한연구 홍누리 안태석 박현수 채한 권영규 김병주 * 부산대학교한의학전문대학원양생기능의학부 Inhibitory Effects of Naeso-san on Pacemaker Potentials in I nterstitial Cells of Cajal of M urine Small I ntestine Noo Ri Hong, Tae Seok Ahn, Hyun Soo Park, Han Chae, Young Kyu Kwon, Byung Joo Kim* Division of Longevity and Biofunctional Medicine, School of Korean Medicine, Pusan National University The purpose of this study was to investigate the effects of Naeso-san in interstitial cells of Cajal (ICCs) in murine small intestine. First, we isolated ICCs from murine small intestine. After that, we cultured these cells for 1 days. The patch-clamp technique was applied on ICCs that formed network-like structures in culture (1 days). Spontaneous rhythms were routinely recorded from cultured ICCs under current-clamp conditions, and the ICCs within networks displayed more robust electrical rhythms (pacemaker potentials). To understand the relationship between Naeso-san and pacemaker activity in ICCs, we examined the effects of Naeso-san on pacemaker potentials of ICCs. In current clamp mode (I = 0), the addition of Naeso-san (10 mg/ml 50 mg/ml) decreased the amplitude and frequency of the pacemaker potentials of ICCs in a dose dependent manner. However, these effects were blocked by intracellular GDPβ S, a G-protein inhibitor, and glibenclamide, a specific ATP-sensitive K+ channels blocker. Pretreatment with SQ-22536, an adenylate cyclase inhibitor, did not block the Naeso-san induced effects, whereas pretreatment with ODQ, a guanylate cyclase inhibitor, or L-NAME, an inhibitor of nitric oxide (NO) synthase blocked the Naeso-san induced effects. Our findings provide insight into unraveling the modulation of Naeso-san in pacemaker potentials of ICCs and developing therapeutic agents against gastrointestinal motility disorders. keywords : Naeso-san, interstitial cells of Cajal, patch-clamp technique, gastrointestinal motility 서론 위장관운동조절제는기능성소화불량이나변비, 과민성대장증상뿐아니라, 당뇨병성위장운동장애, 화학요법으로인한위장운동장애, 소화관운동장애로인한장폐색증, 근긴장성이영양증 (myotonic dystrophy) 환자의위장관운동장애등에사용될수있다. 위장관의운동은교감신경, 부교감신경등과같은외인성신경계, 위장관자체내내인성신경계, 자체내내인성인자및약물등에의해조절된다 1). 위장관운동은위장관평활근의수축으로시발된다. 이평활근은대표적인내장평활근 (visceral smooth muscle) 으로서자동능을보유하고있다. 위장관평활근세포에서기록되는자발적흥분파를서파 (slow wave) 라고부르는데위장관의수축빈도는바로서파의발생빈도에의하여결정된다. 사람에서서 파의발생빈도는부위에따라다르다. 즉위에서는분당 3회, 상부소장에서는 12회이나하부소장으로내려갈수록감소된다 2). 서파의발생기전에대하여는평활근자체라는설 ( 근원설 ) 과특수한간질세포인카할세포 (interstitial cells of Cajal; ICCs) 라는두가지주장이맞서고있으나최근에는카할세포가위장관자발적리듬의향도잡이세포 (pacemaker cell) 이라는많은실험결과가보고되고있다 3,4). 카할세포는평활근과함께간충직 (mesenchyme) 의전구세포에서분화한세포들로위장관평활근및신경망과간극연접 (gap junction) 을통해연결되어있다. 카할세포는세포막의티로신인산화효소 (receptor tyrosine kinase) 인키트 (Kit) 에작용하는 c-kit 유전자를가지고있다 5). 평활근과장관신경계는키트수용체 (Kit receptor) 가존재하지않아 c-kit 항체를이용한세포면역학적방법이카할세포를확인하는데결정적역할을하고있다 3). * Corresponding author Byung Joo Kim, Division of Longevity and Biofunctional Medicine, School of Korean Medicine, Pusan National University, 49, Busandaehak-ro, Mulgeum-eup, Yangsan-si, Gyeongsangnam-do, Korea E-mail : vision@pusan.ac.kr Tel : +82-51-510-8469 Received : 2014/10/08 Revised : 2014/11/17 Accepted : 2014/11/20 c The Korean Society of Korean Pathology, The Korean Society of Korean Physiology pissn 1738-7698 eissn 2288-2529 http://dx.doi.org/10.17208/kjopp.2014.12.28.6.630 Available online at http://society.kisti.re.kr/sv/sv_svjscj03l.do?method=list&poid=ksomp&kojic=drsrdh&svnc=v28n5&menuid=1&subid=13
N. R. Hong et al 631 사람에게서도여러위장관운동성질환이카할세포와크게관련되어있다는많은실험결과들이보고되었다 6-8). 즉카할세포의수가감소하거나형태학적이상이정상적인위장관운동성의리듬을유지하지못함으로인하여운동성장애를일으킨다는것이다. 이와관련된질환들로는염증성장질환 (inflammatory bowel disease), 만성특발성장가짜폐색 (chronic idiopathic intestinal pseudo-obstruction), 식도이완불능증 (achalasia), 선천성거대결장증 (Hirschsprug's disease), 유문협착 (pyloric stenosis), 장폐색 (intestinal obstruction), 항문직장기형 (anorectal malformations) 등이있다 4). 내소산 ( 內消散 ) 은급성위염을치료하는데사용하는처방으로날음식, 찬음식, 딱딱한음식물을먹고위에손상을입어가슴이답답하고아픈증상에응용하여좋은효과가있어급성위염으로음식이위장내에정체되어배가불록하고통증이있는증상을치료하는데사용된다 5). 이와같이내소산은일반적으로위장관계통에문제발생시사용되는시약으로널리알려지고있는데관련된연구로는내소산에의해서흰쥐의위유문부운동성증가가보고되고있으며 5) 위액분비와위궤양치료에효능이있다 9) 고알려지고있다. 또한 항암, 소염, 향균작용을보이고있음이알려지고있다 10,11). 하지만소장운동성에대한내소산의효능은잘알려져있지않다. 따라서, 본실험에서는위장관운동조절기전에관한연구에중요한향도잡이세포인카할세포를이용하여보험용한약제제중에서내소산의카할세포에대한효과와작용기전을알아보아내소산의소장운동성에미치는영향을밝히고자한다. 재료및방법 1. 재료 1) 시약내소산은진피, 반하, 백복령, 지실, 산사육, 신국, 사인, 향부자, 삼릉, 아출, 생강으로이루어져있으며 (Table 1), 아이월드제약회사 (I-WORLD Pharmaceuticals; http://i-pharm.koreasme. com) 에서기증받아사용하였다. 증류수에 0.5 g/ml의농도로녹인후에냉장고에보관사용하였다. 그외다른모든시약은 sigma (Sigma Chemical Co., St. Louis, MO, USA) 에서구입하였고사용직전에원하는농도로만든후세포에투여하였다. 2. 방법 1) 카할세포배양카할세포의배양을위해주로이용되는마우스는 3-5일령 Balb/C를사용하고에테르 (ether) 로마취시킨후개복하여유문륜 (pyloric ring) 에서부터회장에해당하는소장부위를적출하였다. 실온에서크렙스링거중탄산염 (Krebs-Ringer bicarbonate) 용액으로채워진준비용기속에서창자간막가장자리를따라절개하여점막층을제거하고윤상근을노출시킨후분리된소장근육조직을교원질 ( 膠原質 ) 분해효소 (collagenase) (Worthington Biochemical Co., Lakewood, NJ, USA) 1.3 mg/ml, 소혈청알부민 (bovine serum albumin) (Sigma Chemical Co., St. Louis, MO, USA) 2 mg/ml, 트립신저해제 (trypsin inhibitor) (Sigma Chemical Co., St. Louis, MO, USA) 2 mg/ml등이들어있고, Ca 2+ 이들어있지않은행크 (Hank's) 용액에옮긴다음 37 에서 20분간항온소화시킨후진탕시켜세포를분리하였다. 분리된세포들을유리커버글라스위에분주하고, 10분후에간상세포인자 (stem cell factor) (Sigma Chemical Co., St. Louis, MO, USA) 5 ng/ml와 2% 항생 / 항진균제 (antibiotic/antimycotic) (Gibco, Grand Island, NY, USA) 가들어있는 smooth muscle growth medium (Clonetics Corp., San Diego, CA, USA) 용액을분주한후, 37 (95% O2-5% CO 2 ) 배양기에서배양시켰다. 배양된다음날전날배양된용액에서항생 / 항진균제 (antibiotic/antimycotic) 만제외시켜영양액을바꾸어주고실험은배양 1일째후부터시행하였다. 2) 면역형광염색법배양한카할세포를 5분동안아세톤 (acetone) (4 C) 에고정시킨후, 인산완충식염수 (phosphate-buffered saline (PBS; 0.01 M, ph 7.4)) 로희석하고, 0.3% Triton X-100를투여하였다. 그후에실온에서 1% 소혈청알부민 (Bovine Serum Albumin) 을 1시간동안담근후 c-kit 항체 (phycoerythrin-conjugated rat anti-mouse c-kit monoclonal antibody; ebioscience, SanDiego, CA, USA) 를결합하였다. 그후에인산완충식염수로세척한후 2차항체 (fluorescein isothiocyanate (FITC)-coupled donkey anti-rabbit IgG secondary antibody; Jackson Immunoresearch Laboratories, Bar Harbor, MN, U.S.A.) 를결합하였다. 세포는컨포칼현미경 (FV 300 laser scanning confocal microscope; Olympus, Tokyo) 를이용하여자극파장 (excitation wavelength) 인 495 nm에서관찰하였다. 3) 전기생리학적실험배양한카할세포에서막전압을기록하기위해패치클램프 (patch clamp) 실험기법중전세포 (whole cell patch) 방법을이용하였다 12). 전압은패치클램프앰프 (standard patch clamp amplifiers) (Axon Instruments, Foster, CA, USA) 를통해증폭시키며, 나오는신호는컴퓨터모니터및생리적기록기 (Gould 2200, Gould, Valley View, OH, USA) 를통해서관찰하였다. 막전압을기록하는동안세포외관류용액의조성은다음과같다 ( 각수치는 mm 단위임 ): KCl 5, NaCl 135, CaCl 2 1.2, glucose 10, HEPES 10이며 Tris를첨가하여 ph가 7.4가되도록하고, 전극내용액의조성은다음과같이하였다 ( 각수치는 mm 단위임 ): KCl 140, MgCl 2 5, K2ATP 2.7, Na 2GTP 0.1, creatinine phosphate disodium 2.5, HEPES 5, EGTA 0.1이며 Tris를첨가하여 ph가 7.2가되도록하였다. 배양된카할세포에서세포막전류고정법 (Current clamp) 을시행하여자발적으로발생되는내향성향도잡이전압을기록한후내소산의효능을알아보았다. 4) 통계분석대조군과실험군사이의통계학적유의성검정은 SPSS 16.0 (SPSS Inc, Chicago) 의 Independent t-test를사용하였으며유의수준 p<0.05를사용하였다. 실험결과는 mean ± SD 로기재하였다.
632 N. R. Hong et al 결과 1. 내소산의카할세포에대한효과내소산에의한소장카할세포의기능을확인하기전에먼저배양한세포가카할세포인지를 c-kit항체를이용한면역형광염색방법을이용하여확인하였다 (Fig. 1). 소장카할세포의향도잡이기능 (pacemaking activity) 에서내소산에의한조절능을알아보았다. 내소산은농도의존적으로향도잡이기능 (pacemaking activity) 의진폭 (amplitude) 과빈도 (frequency) 는감소하지만안정막전압 (resting membrane potentials) 은거의변화가없음을알수있었다 (Fig. 2). 진폭의변화는 10 mg/ml 내소산에서는거의변화없지만 30 mg/ml 에서는 13.1 ± 2.2 mv (n = 10), 50 mg/ml 에서는 1.7 ± 1.3 mv (n = 8) 정도로진폭이감소되었다 (Fig. 2D). 빈도의변화는 10 mg/ml 내소산에서는거의변화없지만 30 mg/ml 에서는 5.7 ± 1.2 cycles/min (n = 10), 50 mg/ml 에서는 1.1 ± 0.2 cycles/min (n = 8) 정도로빈도가감소되었다 (Fig. 2E). 1.3 ± 1.2 mv (n = 6), 빈도가 2.1 ± 1.0 cycles/min (n = 6) 의반응을보였는데, glibenclamide 투여시에는원래상태로회복이되었다 (Fig. 3B, 3C와 3D). 따라서, 이실험으로내소산은 ATP의존성칼륨이온통로 (ATP sensitive K+ channel) 를통해서카할세포의향도잡이기능 (pacemaking activity) 을억제시킴을알수있었다. Table 1. Amount and Composition of Naeso-san Herb Scientific Name Amount(g) Jin Pi Citri Pericarpium 0.86 g Ban Ha Pinellia ternate(thumb.) Breit 0.68 g Baek Bok Ryung Poria cocos 0.05 g Ji Sil Citrus aurantium 0.78 g San Sa Yuk Crataegus pinnatifida 0.95 g Sin Kuk Massa mddicata fermentata 1.35 g Sa In amomum villosum Lour 0.32 g Hyung Bu Ja Cyperus rotundus L. 0.86 g Sam Ryung Sparganium stoloniferum 0.35 g A Chul Curcuma phaeocaulis 0.24 g Saeng Kang Zingiber officinale 0.26 g Toal amount 6.7 g Fig. 2. Naeso-san decreased the amplitude and frequency of pacemaker potentials in cultured ICCs. (A-C) In current clamp mode (I = 0), the addition of Naeso-san (10 mg/ml - 50 mg/ml) decreased the amplitude and frequency of pacemaker potentials and however, there were no any changes in resting membrane potentials. (D) The histograms summarized the decrease of amplitude with Naeso-san concentration. (E) The histograms summarized the decrease of frequency with Naeso-san concentration. The values were expressed as the mean ± S.D. *P<0.05. **P<0.01. Fig. 1. Cultured interstitial cells of Cajal of the murine small intestine. We identified the cells as ICCs with c-kit immunoreactivity in a single cultured ICC grown for 1 day. Scale bars : 50 μm 2. 내소산의카할세포효과에 ATP의존성칼륨이온통로 (ATP sensitive K+ channel) 의관련성내소산에의한카할세포조절에 ATP의존성칼륨이온통로 (ATP sensitive K+ channel) 의관련성을알아보기위해서억제제인 glibenclamide를사용하여보았다. 일단카할세포에의한향도잡이전압 (pacemaker potentials) 에 glibenclamide는아무런영향을미치지않았다 (Fig. 3A). 내소산 (50 mg/ml) 에의해서는진폭이 Fig. 3. Effects of ATP-sensitive K+ channel inhibitor on pacemaker potentials in cultured ICC from the murine small intestine. (A) Glibenclamide (10 μm) had no effect on the spontaneous pacemaker potentials. (B) The decreased amplitude and frequency of pacemaker potential caused by Naeso-san were prevented by glibenclamide (10 μm). Responses to Naeso-san were summarized in (C) and (D). Bars represented the mean values ± SE. **P< 0.01: significantly different from the untreated control. GBC, glibenclamide.
N. R. Hong et al 633 3. 내소산의카할세포효과에 G protein의관련성내소산에의한카할세포조절에 G protein의관련성을알아보기위해서세포내 G protein의기능을억제시키는것으로알려지고있는 GDPβS를사용하였다. 내소산의카할세포에서의진폭과빈도의억제가 GDPβS (1 mm) 의세포내존재로억제되었다 (Fig. 4). 따라서 G protein이내소산에의한카할세포향도잡이기능 (pacemaking activity) 의억제에관여함을알수있었다. 4. 내소산의카할세포효과에 cyclic GMP 와 nitric oxide의관련성 adenylate cyclase 억제제인 SQ-22536는향도잡이기능 (pacemaking activity) 자체에는아무런효과가없고 (Fig. 5A), 이상태에서내소산투여시내소산단독투여시나타나는현상과동일하게진폭과빈도의감소가나타났다 (Fig. 5B와 5E). 하지만 guanylate cyclase 억제제인 ODQ 투여시에는향도잡이기능 (pacemaking activity) 자체에는아무런효과없고 (Fig. 5C), 내소산에의한진폭과빈도에도아무런효과가나타나지않았다 (Fig. 5D와 5E). 따라서이러한결과는 cyclic AMP는내소산에의한진폭과빈도의감소에관여하지않고 cyclic GMP는관여함을알수있었다. 또산화질소합성효소 (nitric oxide (NO) synthase) 억제제인 L-NAME는향도잡이기능 (pacemaking activity) 자체에는아무런효과없고 (Fig. 6A), 전처치시에도내소산에의한변화가나타나지않아 (Fig. 6B와 6C), NO가내소산에의한변화에관여함을알수있었다. Fig. 4. Effects of GDPβS on the response to Naeso-san. (A) Pacemaker potentials of ICCs exposed to Naeso-san (50 mg/ml) in the presence of GDPβS (1 mm) in the pipette. GDPβS blocked the Naeso-san mediated inhibition of pacemaker potentials. The effects of Naeso-san in the presence of GDPβS in the pipette were summarized in (B) and (C). Bars represented mean values ± S.E. Fig. 6. Effects of a nitric oxide synthase inhibitor on the Naeso-san induced inhibition of pacemaker potentials. (A) L-NAME (10 μm) had no effect on the spontaneous pacemaker potentials. (B) Pacemaker potentials of ICCs exposed Naeso-san in the presence of L-NAME (10 μm). Responses to L-NAME were summarized in (C). Bars represented mean values ± SE. **P< 0.01; significantly different from the untreated control. 고 찰 Fig. 5. Effects of an adenylate cyclase and a guanylate cyclase inhibitor on Naeso-san induced inhibition of pacemaker potentials. (A) SQ-22536 (10 μm) had no effect on the spontaneous pacemaker potentials. (B) Effects of Naeso-san on pacemaker potentials in the presence of SQ-22536 (10 μm). (C) ODQ (10 μm) had no effect on the spontaneous pacemaker potentials. (D) Effects of Naeso-san on pacemaker potentials in the presence of ODQ (10 μm). Responses to SQ-22536 or ODQ were summarized in (E). Bars represented mean values ± SE. **P< 0.01; significantly different from the untreated control. 위장관평활근은신경이나호르몬의외부적자극없이자발적수축을나타내는데이는주기적인세포막전압의변화에기인하며이를서파 (slow wave) 라불린다. 서파는위장관조직절편에서위장관내향도잡이 (pacemaker) 기전이존재함을시사한다 2). 향도잡이 (Pacemaker) 기전에대한연구는오래전부터진행되어왔지만정확한기원에대해서는알려져있지않았다. 그러나최근위장관근육층내에서카할세포 (interstitial cells of Cajal (ICCs)) 이발견되고연구되면서카할세포가위장관의향도잡이 (pacemaker) 세포로작용함이알려져위장관운동생리및병태생리학적연구의주초점이되고있다 3,4). 지금까지알려진카할세포는두가지의기능즉, (1) 향도잡이로서위장관운동성의기본적인리듬을유지하고
634 N. R. Hong et al 있으며 (2) 장관신경계와평활근사이에존재하여신경에의한평활근활동을조절하는중간가교역할을담당하고있다 13,14). 그러나이러한실험결과들은조직에서실험한결과들을바탕으로한것으로카할세포만의정확한생리학적기능들을이해하는데어려움이많았다. 이런어려움을극복하기위하여효소를이용한조직에서단일세포를분리한후형태학적으로카할세포를확인한다음전기생리학적방법을이용하여조직과동일한서파에관련한이온전류들을기록분석하고있다 15). 단일카할세포에서세포막전류고정법 (Current-clamp method) 을시행하면자발적으로향도잡이전압 (pacemaker potentials) 이기록된다 16). 향도잡이전압 (Pacemaker potentials) 의발생기전은세포내 Ca 2+ 저장고인내형질세망에서 IP3를통한주기적 Ca 2+ 분비가미토콘드리아로 Ca 2+ 를유입시켜발생한다. Ca 2+ 저장고인내형질세망에서 IP3를통하여 Ca 2+ 이분비되는기전을차단하는약물과내형질세망으로 Ca 2+ 를재흡수하는기전을차단하는약물에의해서향도잡이전압 (pacemaker potentials) 이억제되며, 미토콘드리아의 uncoupler와호흡연쇄 (respiratory chain) 억제제를사용하면미토콘드리아의 Ca 2+ 농도가감소하면서향도잡이전압 (pacemaker potentials) 이차단된다 17). 이외에도카할세포에는 Ba 2+ 에민감한내부정류칼륨이온 (inward rectifier K+) 통로 18) 와 glibenclamide에반응하는 ATP-의존성 K+ 통로 19) 도존재한다. 내소산은항암및항염효과관련되어많은연구가이루어지고있고 10,11), 위장관과관련되어유문부확장흰쥐의위운동성에대한효능 5) 과위액분비및위궤양에관한연구 9) 외에는별로이루어져있지않다. 또한위장관운동조절에중요한카할세포에서의내소산에대한연구는전혀이루어지지않고있다. 본실험에서는위장관질환치료에사용되는내소산에서위장관운동향도잡이 (pacemaker) 인카할세포에서의효능을알아보았다. 카할세포에의한향도잡이전압 (pacemaker potentials) 에서내소산에서는안정막전압은큰변화없으며진폭과빈도가감소됨을알수있었고 (Fig. 2), 내소산에의한반응에 ATP 의존성 K+ 이온통로가관여함을알수있었다 (Fig. 3). 세포내 G protein이내소산의반응에영향을미치고 (Fig. 4), cyclic GMP와 NO가관여함을알수있었다 (Fig. 5와 6). 결국내소산은카할세포의안정막전압의큰변화없이진폭과빈도수를억제하여위장관수축력억제를일으킨다고사료되고이때세포내 G protein-cyclic GMP-NO 기전이관여한다고생각된다. 앞으로는이런반응에관여하는다양한세포내기전들과 in vivo 상태에서의내소산에의한위장관운동성변화에대하서연구를진행할예정이다. 또한카할세포에의한향도잡이변화에는비선택성양이온통로인일과성수용체전압 (TRPM7; transient receptor potential melastatin 7) 이온통로 12) 와칼슘에의해활성화되는염소 (Ca 2+ -activated Cl - ) 이온통로 20) 가관여하고있음이밝혀졌다. 따라서내소산이이들이온통로에는어떤효과를보이는지를확인하고실제로내소산이카할세포의기능조절에이들이온통로를조절하고있는지를연구할예정이다. 카할세포의바이오마커로는 c-kit 21) 과칼슘에의해활성화되는염소이온통로 22) 가알려지고있는데내소산에의한 in vivo 실험시이러한바이오마커의카할세포세포막발현에미치는영향도큰의미있는실험주제가될것 으로생각된다. 그동안다양한한약제제를이용하여카할세포에서의반응을관찰하였다. 삼출견비탕 23), 이중탕 24), 소청룡탕 25), 삼황사심탕 26) 에의해서는카할세포의흥분성을유발하는즉, 탈분극 (depolarization) 시키는방향으로효능이나타나위장관운동성을증가시키는쪽으로나타났다. 삼출견비탕 23) 과이중탕 24) 은세포내외의칼슘과세포막 G-단백질및 phospholipase C가관여하여탈분극을시키고, 소청룡탕 25) 과삼황사심탕 26) 은 5-HT 3번과 4번수용체를통한 Mitogen-Activated Protein Kinases에의해서효과가나타났다. 하지만, 내소산에의한반응은안정막전압은큰변화없지만진폭과빈도가감소함에따라위장관운동성을억제시키는방향으로효과가나타나는것으로사료된다. 따라서내소산은위장관경련, 통증, 일시적기능억제나다른위장관운동이상과관련된질환에대한치료제개발에관여할수있음을보여준다. 결론 현재카할세포에대한생리약리학적실험결과들은극히드물다. 카할세포에의한한약제들의약리학적연구는위장관운동관련치료제의개발과연관하여특히중요하다고생각된다. 현재까지알려진위장관운동과관련된질환은카할세포의수가감소하거나기능장애에의한향도잡이기능의손실로인한위장관운동장애로추정하고있다. 따라서카할세포에의한한약제들의약리학적연구는위장관운동관련치료제의개발과연관하여특히중요할것으로사료된다. 본연구에서내소산은위장관카할세포에서 ATP 의존성 K+ 이온통로의존적으로 G protein-cyclic GMP-NO 기전이관여하여향도잡이기능을억제하고있다. 따라서내소산은위장관운동조절치료제로서개발가능성있는한약제제라생각된다. 감사의글 본연구는보건복지부한의약선도기술개발사업의지원에의하여이루어진것임 (HI12C1886;B120008). References 1. Jun, J.Y. Interstitial Cells of Cajal. Korean J Gastrointest Motil. 7: 70-72, 2001. 2. Park, K.J. Interstitial Cells of Cajal and GI Motility. J Neurogastroenterol Motil. 10: 93-99, 2009. 3. Huizinga, J.D., Thuneberg, L., Kluppel, M., Malysz, H.B. The W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373: 347-349, 1995. 4. Sanders, K.M., Tamas, O., Kho, S.D., Torihashi, S., Ward, S.M. Deveolpment of interstitial cells of cajal. Neurogastrointestinal motility 11: 3-11, 1999. 5. Kim, J., Yoon, S. Effect of Naesosan on Gastric Motility
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