Changes of c-kit and scf mRNA and Protein in Guinea Pig Gallbladder Fed on High Cholesterol Diet_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

FAN Ying ¹ , WU Shuodong ¹ , FU Beibei ² , YIN Zhenhua ¹

1. Department of The Second General Surgery， Shengjing Hospital， China Medical University， Shenyang 110004， Liaoning Province， China;;
2. Department of Paediatrics， Shengjing Hospital， China Medical University， Shenyang 110004， Liaoning Province， China;

Corresponding author：

Keywords：

Cholesterol gallstones; c-kit gene; Stem cell factor; RT-PCR; Western blot

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Objective 　To discuss the changes of c-kit/scf mRNA and protein in guinea pig gallbladder fed on high cholesterol diet.
Methods 　Twenty guinea pigs were divided into two equal groups of 10 each：the control group and lithogenic group. Normal diet and high cholesterol diet was given to each group respectively. The period of stone permeation was six weeks. RT-PCR and Western blot were used to determin the expressions of c-kit and scf mRNA and protein.
Results 　RT-PCR results showed that the expressions of c-kit mRNA（t=6.985，P＜0.01） and scf mRNA （t=6.028， P＜0.01）decreased significantly in lithogenic group compared with the control group. Western blot results showed that the expressions of c-kit protein （t=10.256， P＜0.01） and scf protein （t=9.586， P＜0.01）decreased significantly in lithogenic group compared with the control group.
Conclusions 　The expressions of c-kit/scf mRNA and protein decrease during the formation of cholesterol gallstones in guinea pigs fed on high cholesterol diet. Inhibition of c-kit/scf pathway may play a role in the formation of cholesterol gallstones.

Citation： FAN Ying,WU Shuodong,FU Beibei,YIN Zhenhua. Changes of c-kit and scf mRNA and Protein in Guinea Pig Gallbladder Fed on High Cholesterol Diet. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2012, 19(10): 1063-1068. doi: Copy

1.	Wang HH， Portincasa P， Liu M， et al. Effect of gallbladder hypomotility on cholesterol crystallization and growth in CCK-deficient mice[J]. Biochim Biophys Acta， 2010， 1801(2)：138-146.
2.	Torihashi S， Ward SM， Nishikawa S， et al. c-kit-dependentdevelopment of interstitial cells and electrical activity in the murinegastrointestinal tract[J]. Cell Tissue Res， 1995， 280(1)：97-111.
3.	Thuneberg L. Interstitial cells of Cajal：intestinal pacemaker cells?[J]. Adv Anat Embryol Cell Biol， 1982， 71：1-130.
4.	Mazzia C， Porcher C， Julé Y， et al. Ultrastructural study of relationships between c-kit immunoreactive interstitial cells and other cellular elements in the human colon[J]. Histochem Cell Biol， 2000， 113(5)：401-411.
5.	Horiguchi K， Sanders KM， Ward SM. Enteric motor neurons form synaptic-like junctions with interstitial cells of Cajal in the canine gastric antrum[J]. Cell Tissue Res， 2003， 311(3)：299-313.
6.	Wang XY， Sanders KM， Ward SM. Intimate relationshipbetween interstitial cells of Cajal and enteric nerves in the guinea-pig small intestine[J]. Cell Tissue Res， 1999， 295(2)：247-256.
7.	Ward SM， Beckett EA， Wang X， et al. Interstitial cells ofCajal mediate cholinergic neurotransmission from enteric motorneurons[J]. J Neurosci， 2000， 20(4)：1393-1403.
8.	Sanders KM， Ward SM. Interstitial cells of Cajal：a new perspe-ctive on smooth muscle function[J]. J Physiol， 2006， 576(Pt 3)：721-726.
9.	Hinescu ME， Ardeleanu C， Gherghiceanu M， et al. Interstitial Cajal-like cells in human gallbladder[J]. J Mol Histol， 2007， 38(4)：275-284.
10.	Lavoie B， Balemba OB， Nelson MT， et al. Morphological and physiological evidence for interstitial cell of Cajal-like cells in the guinea pig gallbladder[J]. J Physiol， 2007， 579(Pt 2)：487-501.
11.	Sun X， Yu B， Xu L ，et al. Interstitial cells of Cajal in the murine gallbladder[J]. Scand J Gastroenterol， 2006， 41(10)：1218-1226.
12.	Hu WM， Luo HS， Ding XW， et al. Expression of c-kit messenger ribonucleic acid and c-kit protein in the gallbladders in guinea pigs of high cholesterol diet[J]. Dig Dis Sci， 2009， 54(8)：1651-1655.
13.	Shaffer EA. Review article：control of gall-bladder motor function[J]. Aliment Pharmacol Ther，2000， 14(Suppl 2)：2-8.
14.	Taylor AM， Galli SJ， Coleman JW. Stem-cell factor， the kit ligand， induces direct degranulation of rat peritoneal mast cells in vitro and in vivo：dependence of the in vitro effect on period of culture and comparisons of stem-cell factor with other mast cell-activating agents[J]. Immunology，1995，86(3)：427-433.
15.	Kitamura Y， Hirota S， Nishida T. A loss-of-function mutation of c-kit results in depletion of mast cells and interstitial cells of Cajal，while its gain-of-function mutation results in their oncogenesis[J]. Mutat Res， 2001， 477(1-2)：165-171.
16.	Fan Y， Wu SD， Fu BB. Effect of intestinal transit on the formation of cholesterol gallstones in hamsters[J]. Hepatobiliary Pancreat Dis Int， 2007， 6(5)：513-515.
17.	Xu D， Yu BP， Luo HS， et al. Control of gallbladder contractions by cholecystokinin through cholecystokinin-A receptors on gallbladder interstitial cells of Cajal[J]. World J Gastroenterol， 2008， 14(18)：2882-2887.
18.	Torihashi S， Yokoi K， Nagaya H， et al. New monoclonal antibody (AIC) identifies interstitial cells of Cajal in the musculature of the mouse gastrointestinal tract[J]. Auton Neurosci， 2004， 113(1-2)：16-23.
19.	Ward SM， Sanders KM. Physiology and pathophysiology of the interstitial cell of Cajal：from bench to bedside.Ⅰ. Functional development and plasticity of interstitial cells of Cajal networks[J]. Am J Physiol Gastrointest Liver Physiol， 2001， 281(3)：G602-G611.
20.	Torihashi S， Nishi K， Tokutomi Y， et al. Blockade of kit signaling induces transdifferentiation of interstitial cells of Cajal to a smooth muscle phenotype[J]. Gastroenterology， 1999， 117(1)：140-148.
21.	Albertí E， Mikkelsen HB， Wang XY， et al. Pacemaker activity and inhibitory neurotransmission in the colon of Ws/Ws mutant rats[J]. Am J Physiol Gastrointest Liver Physiol， 2007， 292(6)：G1499-G1510.
22.	Lyford GL， He CL， Soffer E， et al. Pan-colonic decrease in interstitial cells of Cajal in patients with slow transit constipation[J]. Gut， 2002， 51(4)：496-501.
23.	Battaglia E， Bassotti G， Bellone G， et al. Loss of interstitial cells of Cajal network in severe idiopathic gastroparesis[J]. World J Gastroenterol， 2006， 12(38)：6172-6177.
24.	Porcher C， Baldo M， Henry M， et al. Deficiency of interstitial cells of Cajal in the small intestine of patients with Crohn’s disease[J]. Am J Gastroenterol， 2002， 97(1)：118-125.
25.	Orlic D， Kajstura J， Chimenti S， et al. Mobilized bone marrow cells repair the infarcted heart， improving function and survival[J]. Proc Natl Acad Sci U S A， 2001， 98(18)：10344-10349.
26.	Ward SM， Ordög T， Bayguinov JR， et al. Development of interstitial cells of Cajal and pacemaking in mice lacking enteric nerves[J]. Gastroenterology， 1999， 117(3)：584-594.
27.	Ward SM， Burns AJ， Torihashi S， et al. Impaired development of interstitial cells and intestinal electrical rhythmicity in steel mutants[J]. Am J Physiol Cell Physiol， 1995， 269(6 Pt 1)：C1577-C1585.
28.	Rich A， Miller SM， Gibbons SJ， et al. Local presentation of steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture[J]. Am J Physiol Gastrointest Liver Physiol， 2003， 284(2)：G313-G320.
29.	Nakahara M， Isozaki K， Vanderwinden JM， et al. Dose-dependent and time-limited proliferation of cultured murine interstitial cells of Cajal in response to stem cell factor[J]. Life Sci， 2002， 70(20)：2367-2376.

1. Wang HH， Portincasa P， Liu M， et al. Effect of gallbladder hypomotility on cholesterol crystallization and growth in CCK-deficient mice[J]. Biochim Biophys Acta， 2010， 1801(2)：138-146.
2. Torihashi S， Ward SM， Nishikawa S， et al. c-kit-dependentdevelopment of interstitial cells and electrical activity in the murinegastrointestinal tract[J]. Cell Tissue Res， 1995， 280(1)：97-111.
3. Thuneberg L. Interstitial cells of Cajal：intestinal pacemaker cells?[J]. Adv Anat Embryol Cell Biol， 1982， 71：1-130.
4. Mazzia C， Porcher C， Julé Y， et al. Ultrastructural study of relationships between c-kit immunoreactive interstitial cells and other cellular elements in the human colon[J]. Histochem Cell Biol， 2000， 113(5)：401-411.
5. Horiguchi K， Sanders KM， Ward SM. Enteric motor neurons form synaptic-like junctions with interstitial cells of Cajal in the canine gastric antrum[J]. Cell Tissue Res， 2003， 311(3)：299-313.
6. Wang XY， Sanders KM， Ward SM. Intimate relationshipbetween interstitial cells of Cajal and enteric nerves in the guinea-pig small intestine[J]. Cell Tissue Res， 1999， 295(2)：247-256.
7. Ward SM， Beckett EA， Wang X， et al. Interstitial cells ofCajal mediate cholinergic neurotransmission from enteric motorneurons[J]. J Neurosci， 2000， 20(4)：1393-1403.
8. Sanders KM， Ward SM. Interstitial cells of Cajal：a new perspe-ctive on smooth muscle function[J]. J Physiol， 2006， 576(Pt 3)：721-726.
9. Hinescu ME， Ardeleanu C， Gherghiceanu M， et al. Interstitial Cajal-like cells in human gallbladder[J]. J Mol Histol， 2007， 38(4)：275-284.
10. Lavoie B， Balemba OB， Nelson MT， et al. Morphological and physiological evidence for interstitial cell of Cajal-like cells in the guinea pig gallbladder[J]. J Physiol， 2007， 579(Pt 2)：487-501.
11. Sun X， Yu B， Xu L ，et al. Interstitial cells of Cajal in the murine gallbladder[J]. Scand J Gastroenterol， 2006， 41(10)：1218-1226.
12. Hu WM， Luo HS， Ding XW， et al. Expression of c-kit messenger ribonucleic acid and c-kit protein in the gallbladders in guinea pigs of high cholesterol diet[J]. Dig Dis Sci， 2009， 54(8)：1651-1655.
13. Shaffer EA. Review article：control of gall-bladder motor function[J]. Aliment Pharmacol Ther，2000， 14(Suppl 2)：2-8.
14. Taylor AM， Galli SJ， Coleman JW. Stem-cell factor， the kit ligand， induces direct degranulation of rat peritoneal mast cells in vitro and in vivo：dependence of the in vitro effect on period of culture and comparisons of stem-cell factor with other mast cell-activating agents[J]. Immunology，1995，86(3)：427-433.
15. Kitamura Y， Hirota S， Nishida T. A loss-of-function mutation of c-kit results in depletion of mast cells and interstitial cells of Cajal，while its gain-of-function mutation results in their oncogenesis[J]. Mutat Res， 2001， 477(1-2)：165-171.
16. Fan Y， Wu SD， Fu BB. Effect of intestinal transit on the formation of cholesterol gallstones in hamsters[J]. Hepatobiliary Pancreat Dis Int， 2007， 6(5)：513-515.
17. Xu D， Yu BP， Luo HS， et al. Control of gallbladder contractions by cholecystokinin through cholecystokinin-A receptors on gallbladder interstitial cells of Cajal[J]. World J Gastroenterol， 2008， 14(18)：2882-2887.
18. Torihashi S， Yokoi K， Nagaya H， et al. New monoclonal antibody (AIC) identifies interstitial cells of Cajal in the musculature of the mouse gastrointestinal tract[J]. Auton Neurosci， 2004， 113(1-2)：16-23.
19. Ward SM， Sanders KM. Physiology and pathophysiology of the interstitial cell of Cajal：from bench to bedside.Ⅰ. Functional development and plasticity of interstitial cells of Cajal networks[J]. Am J Physiol Gastrointest Liver Physiol， 2001， 281(3)：G602-G611.
20. Torihashi S， Nishi K， Tokutomi Y， et al. Blockade of kit signaling induces transdifferentiation of interstitial cells of Cajal to a smooth muscle phenotype[J]. Gastroenterology， 1999， 117(1)：140-148.
21. Albertí E， Mikkelsen HB， Wang XY， et al. Pacemaker activity and inhibitory neurotransmission in the colon of Ws/Ws mutant rats[J]. Am J Physiol Gastrointest Liver Physiol， 2007， 292(6)：G1499-G1510.
22. Lyford GL， He CL， Soffer E， et al. Pan-colonic decrease in interstitial cells of Cajal in patients with slow transit constipation[J]. Gut， 2002， 51(4)：496-501.
23. Battaglia E， Bassotti G， Bellone G， et al. Loss of interstitial cells of Cajal network in severe idiopathic gastroparesis[J]. World J Gastroenterol， 2006， 12(38)：6172-6177.
24. Porcher C， Baldo M， Henry M， et al. Deficiency of interstitial cells of Cajal in the small intestine of patients with Crohn’s disease[J]. Am J Gastroenterol， 2002， 97(1)：118-125.
25. Orlic D， Kajstura J， Chimenti S， et al. Mobilized bone marrow cells repair the infarcted heart， improving function and survival[J]. Proc Natl Acad Sci U S A， 2001， 98(18)：10344-10349.
26. Ward SM， Ordög T， Bayguinov JR， et al. Development of interstitial cells of Cajal and pacemaking in mice lacking enteric nerves[J]. Gastroenterology， 1999， 117(3)：584-594.
27. Ward SM， Burns AJ， Torihashi S， et al. Impaired development of interstitial cells and intestinal electrical rhythmicity in steel mutants[J]. Am J Physiol Cell Physiol， 1995， 269(6 Pt 1)：C1577-C1585.
28. Rich A， Miller SM， Gibbons SJ， et al. Local presentation of steel factor increases expression of c-kit immunoreactive interstitial cells of Cajal in culture[J]. Am J Physiol Gastrointest Liver Physiol， 2003， 284(2)：G313-G320.
29. Nakahara M， Isozaki K， Vanderwinden JM， et al. Dose-dependent and time-limited proliferation of cultured murine interstitial cells of Cajal in response to stem cell factor[J]. Life Sci， 2002， 70(20)：2367-2376.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Changes of c-kit and scf mRNA and Protein in Guinea Pig Gallbladder Fed on High Cholesterol Diet

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