豚鼠胰腺内telocytes形态学特点及甲磺酸伊马替尼对telocytes的干预作用
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摘要
目的对豚鼠胰腺组织特络细胞(telocytes,TCs)的形态特征进行观察与分析,并探讨甲磺酸伊马替尼对TCs的干预作用。方法以健康9周龄豚鼠胰腺为研究对象,利用免疫组化染色对TCs免疫标志(c-kit/CD117、vimentin)进行观察分析。透射电镜观察TCs的超微结构。建立甲磺酸伊马替尼干预模型:豚鼠54只,雌雄不限,分正常对照组、用药组(7、14、21、28 d)、停药组(W7、W14、W21、W28 d);免疫组化染色观察用/停药后TCs变化;Western blot观察用/停药后胰腺组织内CD117、vimentin蛋白表达。结果免疫组化染色显示:胰腺内TCs位于结缔组织内,细胞呈现三角形及长梭形等多种形态,有一至多条长而细的细胞突起,突起包绕腺泡,并相连形成网络结构;电镜超微结构显示:TCs有念珠状的细长突起,核质比较高,胞质内含有少量的线粒体、滑面内质网等;甲磺酸伊马替尼干预后,TCs突起变短,网络结构减少,停药后逐渐恢复;用药后胰腺组织内c-kit/CD117和vimentin蛋白表达降低,停药后逐渐恢复,差异有统计学意义(P<0.05)。结论豚鼠胰腺内存在梭形有念珠状突起的TCs,TCs包绕腺泡形成网络结构;甲磺酸伊马替尼可影响胰腺内TCs的形态及蛋白表达。
Objective To observe the morphological characteristics of telocytes(TCs) in the pancreas of guinea pigs,and determine the effect of imatinib mesylate on TCs. Methods Immunohistochemical staining was used to observe the immune markers,c-kit,CD117 and vimentin in the TCs from the pancreas of guinea pigs(9 weeks old). Transmission electron microscopy(TEM) was used to observed the ultrastructure of TCs. Fifty-four guinea pigs of either sex were randomly divided into normal control group,imatinib mesylate treatment groups(7,14,21 and 28 d) and drug withdrawal groups(in 7,14,21 and 28 d after withdraw).After treatment and drug withdrawal,immunohistochemical staining was employed to observe the changes in the TCs,and Western blotting was adopted to detect the expression changes of c-kit,CD117 and vimentin.Results Immunohistochemical staining showed that pancreatic TCs were located in the connective tissues,and the cells were in triangular,long spindle and other shapes with one or more long and fine cell processes,protruding around the acini and connected into network structures. ETM results displayed that the TCs had slender moniliform processes,higher nucleus/cytoplasm ratio,and mitochondria and smooth endoplasmic reticula in the cytoplasm. Imatinib mesylate treatment induced shorter cell processes and less network structures,but the phenomena were recovered after drug withdrawal. The drug intervention also decreased the protein levels of c-kit/CD117 and vimentin,and the levels were also restored after the withdrawal(P < 0. 05).Conclusion In the pancreas of guinea pigs,there are spindle-shaped TCs with moniliform processes,and the TCs protrude around the acini and connect into network structures. Imatinib mesylate affects the morphology and protein expression in the TCs.
Objective To observe the morphological characteristics of telocytes(TCs) in the pancreas of guinea pigs,and determine the effect of imatinib mesylate on TCs. Methods Immunohistochemical staining was used to observe the immune markers,c-kit,CD117 and vimentin in the TCs from the pancreas of guinea pigs(9 weeks old). Transmission electron microscopy(TEM) was used to observed the ultrastructure of TCs. Fifty-four guinea pigs of either sex were randomly divided into normal control group,imatinib mesylate treatment groups(7,14,21 and 28 d) and drug withdrawal groups(in 7,14,21 and 28 d after withdraw).After treatment and drug withdrawal,immunohistochemical staining was employed to observe the changes in the TCs,and Western blotting was adopted to detect the expression changes of c-kit,CD117 and vimentin.Results Immunohistochemical staining showed that pancreatic TCs were located in the connective tissues,and the cells were in triangular,long spindle and other shapes with one or more long and fine cell processes,protruding around the acini and connected into network structures. ETM results displayed that the TCs had slender moniliform processes,higher nucleus/cytoplasm ratio,and mitochondria and smooth endoplasmic reticula in the cytoplasm. Imatinib mesylate treatment induced shorter cell processes and less network structures,but the phenomena were recovered after drug withdrawal. The drug intervention also decreased the protein levels of c-kit/CD117 and vimentin,and the levels were also restored after the withdrawal(P < 0. 05).Conclusion In the pancreas of guinea pigs,there are spindle-shaped TCs with moniliform processes,and the TCs protrude around the acini and connect into network structures. Imatinib mesylate affects the morphology and protein expression in the TCs.
引文
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[18]KOSTIN S.Cardiac telocytes in normal and diseased hearts[J].Semin Cell Dev Biol,2016,55:22-30.DOI:10.1016/j.semcdb.2016.02.023.
[19]LIU J,CAO Y,SONG Y,et al.Telocytes in liver[J].Curr Stem Cell Res Ther,2016,11(5):415-419.DOI:10.2174/1574888x10666150630112035.
[20]CRETOIU S M,POPESCU L M.Telocytes revisited[J].Biomol Concepts,2014,5(5):353-369.DOI:10.1515/bmc-2014-0029.
[21]RATAJCZAK M Z,RATAJCZAK D,PEDZIWIATR D.Extracellular microvesicles(Ex MVs)in cell to cell communication:A role of telocytes[J].Advances in Experimental Medicine and Biology,2016,913:41-49.DOI:10.1007/978-981-10-1061-3_3.
[22]SHIMOJIMA N,NAKAKI T,MORIKAWA Y,et al.Imatinib blocks spontaneous mechanical activities in the adult mouse small intestine:Possible inhibition of c-Kit signaling[J].Pharmacology,2005,74(2):95-99.DOI:10.1159/000084021.
[23]DRUKER B.Imatinib(Gleevec)as a paradigm of targeted cancer therapies[J].Keio J Med,2010,59(1):1-3.DOI:10.2302/kjm.59.1.
[24]ORDG T.Interstitial cells of Cajal in diabetic gastroenteropathy[J].Neurogastroenterol Motil,2008,20(1):8-18.DOI:10.1111/j.1365-2982.2007.01056.x.
[2]CORRADI L S,JESUS M M,FOCHI R A,et al.Structural and ultrastructural evidence for telocytes in prostate stroma[J].J Cell Mol Med,2013,17(3):398-406.DOI:10.1111/jcmm.12021.
[3]CHEN X,ZHENG Y,MANOLE C G,et al.Telocytes in human oesophagus[J].J Cell Mol Med,2013,17(11):1506-1512.DOI:10.1111/jcmm.12149.
[4]ZHANG H Q,LU S S,XU T,et al.Morphological evidence of telocytes in mice aorta[J].Chin Med J,2015,128(3):348-352.DOI:10.4103/0366-6999.150102.
[5]CHANG Y,LI C,GAN L,et al.Telocytes in the Spleen[J].PLo S ONE,2015,10(9):e0138851.DOI:10.1371/journal.pone.0138851.
[6]RUSU M C,LORETO C,MNOIU V S.Network of telocytes in the temporomandibular joint disc of rats[J].Acta Histochem,2014,116(4):663-668.DOI:10.1016/j.acthis.2013.12.005.
[7]VANNUCCHI M G,TRAINI C.Interstitial cells of Cajal and telocytes in the gut:twins,related or simply neighbor cells[J].Biomol Concepts,2016,7(2):93-102.DOI:10.1515/bmc-2015-0034.
[8]KOSTIN S,POPESCU L M.A distinct type of cell in myocardium interstitial cajal-like cells(ICLCs)[J].J Cell Mol Med,2009,13(2):295-308.DOI:10.1111/j.1582-4934.2008.00668.x.
[9]POPESCU L M,FAUSSONE-PELLEGRINI M S.TELOCYTES-a case of serendipity:the winding way from Interstitial Cells of Cajal(ICC),via Interstitial Cajal-Like Cells(ICLC)to TELOCYTES[J].J Cell Mol Med,2010,14(4):729-740.DOI:10.1111/j.1582-4934.2010.01059.x.
[10]ZHENG Y,BAI C,WANG X.Telocyte morphologies and potential roles in diseases[J].J Cell Physiol,2012,227(6):2311-2317.DOI:10.1002/jcp.23022.
[11]VANNUCCHI M G,FAUSSONE-PELLEGRINI M S.The Telocyte Subtypes[J].Adv Exp Med Biol,2016,913:115-126.DOI:10.1007/978-981-10-1061-3_7.
[12]NICOLESCU M I,POPESCU L M.Telocytes in the interstitium of human exocrine pancreas:ultrastructural evidence[J].Pancreas,2012,41(6):949-956.DOI:10.1097/MPA.0b013e31823fbded.
[13]MEI F,HAN J,HUANG Y,et al.Plasticity of interstitial cells of cajal:a study in the small intestine of adult Guinea pigs[J].Anat Rec(Hoboken),2009,292(7):985-993.DOI:10.1002/ar.20928.
[14]LORINCZ A,REDELMAN D,HORVTH V J,et al.Progenitors of interstitial cells of cajal in the postnatal murine stomach[J].Gastroenterology,2008,134(4):1083-1093.DOI:10.1053/j.gastro.2008.01.036.
[15]DAZ-FLORES L,GUTIRREZ R,DAZ-FLORES L Jr,et al.Behaviour of telocytes during physiopathological activation[J].Semin Cell Dev Biol,2016,55:50-61.DOI:10.1016/j.semcdb.2016.01.035.
[16]CISMASIU V B,POPESCU L M.Telocytes transfer extracellular vesicles loaded with microRNAs to stem cells[J].J Cell Mol Med,2015,19(2):351-358.DOI:10.1111/jcmm.12529.
[17]WANG J,JIN M,MA W H,et al.The history of telocyte discovery and understanding[J].Adv Exp Med Biol,2016,913:1-21.DOI:10.1007/978-981-10-1061-3_1.
[18]KOSTIN S.Cardiac telocytes in normal and diseased hearts[J].Semin Cell Dev Biol,2016,55:22-30.DOI:10.1016/j.semcdb.2016.02.023.
[19]LIU J,CAO Y,SONG Y,et al.Telocytes in liver[J].Curr Stem Cell Res Ther,2016,11(5):415-419.DOI:10.2174/1574888x10666150630112035.
[20]CRETOIU S M,POPESCU L M.Telocytes revisited[J].Biomol Concepts,2014,5(5):353-369.DOI:10.1515/bmc-2014-0029.
[21]RATAJCZAK M Z,RATAJCZAK D,PEDZIWIATR D.Extracellular microvesicles(Ex MVs)in cell to cell communication:A role of telocytes[J].Advances in Experimental Medicine and Biology,2016,913:41-49.DOI:10.1007/978-981-10-1061-3_3.
[22]SHIMOJIMA N,NAKAKI T,MORIKAWA Y,et al.Imatinib blocks spontaneous mechanical activities in the adult mouse small intestine:Possible inhibition of c-Kit signaling[J].Pharmacology,2005,74(2):95-99.DOI:10.1159/000084021.
[23]DRUKER B.Imatinib(Gleevec)as a paradigm of targeted cancer therapies[J].Keio J Med,2010,59(1):1-3.DOI:10.2302/kjm.59.1.
[24]ORDG T.Interstitial cells of Cajal in diabetic gastroenteropathy[J].Neurogastroenterol Motil,2008,20(1):8-18.DOI:10.1111/j.1365-2982.2007.01056.x.