胃肠病学
胃腸病學
위장병학
CHINESE JOURNAL OF GASTROENTEROLOGY
2014年
12期
719-724
,共6页
王庆娥%朱滢%王云%孙晓萌%林琳
王慶娥%硃瀅%王雲%孫曉萌%林琳
왕경아%주형%왕운%손효맹%림림
糖尿病%糖基化终末产物%肌细胞,平滑肌%超微结构%收缩蛋白质类
糖尿病%糖基化終末產物%肌細胞,平滑肌%超微結構%收縮蛋白質類
당뇨병%당기화종말산물%기세포,평활기%초미결구%수축단백질류
Diabetes Mellitus%Advanced Glycation End Products%Myocytes,Smooth Muscle%Ultrastructure%Contractile Proteins
背景：糖尿病性胃轻瘫（ DGP）与平滑肌细胞（ SMC）病变有关，糖基化终末产物（ AGEs）在此病变中的作用尚未完全明确。目的：探讨糖尿病（ DM）患者胃SMC超微结构的变化，分析收缩蛋白表达与AGEs的关系。方法：选取2012年7月至2012年12月南京医科大学第一附属医院因胃肿瘤行胃切除术的30例患者的全层胃组织，DM和非DM患者各15例，分别作为DM组和对照组。以透射电子显微镜观察SMC超微结构，以蛋白质印迹法检测胃肌层收缩蛋白肌球蛋白重链（ MHC）、α-肌动蛋白（α-actin）、钙调节蛋白（ calponin）表达和AGEs重要组分Nε-羧甲基赖氨酸（ CML）表达，以荧光定量PCR 检测MHC、α-actin、calponin mRNA表达。以Pearson相关系数分析各收缩蛋白mRNA表达与CML蛋白表达的相关性。结果：DM患者胃肌层SMC超微结构明显异常，表现为缝隙连接破坏、胞质内线粒体肿胀、出现脂褐素、细胞膜小凹数量增加和细胞膜外基层增厚。DM组胃肌层MHC、α-actin、calponin蛋白和mRNA表达水平均较对照组显著降低（P均﹤0.01），CML蛋白表达水平则显著高于对照组（P﹤0.01）。DM组胃肌层MHC、calponin mRNA表达与CML蛋白表达呈负相关（r=-0.59，P=0.02；r=-0.63，P=0.01），α-actin mRNA表达则与CML蛋白表达无明显相关性（r=-0.49，P=0.06）。结论：DM患者存在胃肌层SMC超微结构破坏，肌层收缩蛋白减少、AGEs增多且两者呈负相关。上述变化可能影响SMC收缩功能，进而参与DM胃动力障碍的发生。
揹景：糖尿病性胃輕癱（ DGP）與平滑肌細胞（ SMC）病變有關，糖基化終末產物（ AGEs）在此病變中的作用尚未完全明確。目的：探討糖尿病（ DM）患者胃SMC超微結構的變化，分析收縮蛋白錶達與AGEs的關繫。方法：選取2012年7月至2012年12月南京醫科大學第一附屬醫院因胃腫瘤行胃切除術的30例患者的全層胃組織，DM和非DM患者各15例，分彆作為DM組和對照組。以透射電子顯微鏡觀察SMC超微結構，以蛋白質印跡法檢測胃肌層收縮蛋白肌毬蛋白重鏈（ MHC）、α-肌動蛋白（α-actin）、鈣調節蛋白（ calponin）錶達和AGEs重要組分Nε-羧甲基賴氨痠（ CML）錶達，以熒光定量PCR 檢測MHC、α-actin、calponin mRNA錶達。以Pearson相關繫數分析各收縮蛋白mRNA錶達與CML蛋白錶達的相關性。結果：DM患者胃肌層SMC超微結構明顯異常，錶現為縫隙連接破壞、胞質內線粒體腫脹、齣現脂褐素、細胞膜小凹數量增加和細胞膜外基層增厚。DM組胃肌層MHC、α-actin、calponin蛋白和mRNA錶達水平均較對照組顯著降低（P均﹤0.01），CML蛋白錶達水平則顯著高于對照組（P﹤0.01）。DM組胃肌層MHC、calponin mRNA錶達與CML蛋白錶達呈負相關（r=-0.59，P=0.02；r=-0.63，P=0.01），α-actin mRNA錶達則與CML蛋白錶達無明顯相關性（r=-0.49，P=0.06）。結論：DM患者存在胃肌層SMC超微結構破壞，肌層收縮蛋白減少、AGEs增多且兩者呈負相關。上述變化可能影響SMC收縮功能，進而參與DM胃動力障礙的髮生。
배경：당뇨병성위경탄（ DGP）여평활기세포（ SMC）병변유관，당기화종말산물（ AGEs）재차병변중적작용상미완전명학。목적：탐토당뇨병（ DM）환자위SMC초미결구적변화，분석수축단백표체여AGEs적관계。방법：선취2012년7월지2012년12월남경의과대학제일부속의원인위종류행위절제술적30례환자적전층위조직，DM화비DM환자각15례，분별작위DM조화대조조。이투사전자현미경관찰SMC초미결구，이단백질인적법검측위기층수축단백기구단백중련（ MHC）、α-기동단백（α-actin）、개조절단백（ calponin）표체화AGEs중요조분Nε-최갑기뢰안산（ CML）표체，이형광정량PCR 검측MHC、α-actin、calponin mRNA표체。이Pearson상관계수분석각수축단백mRNA표체여CML단백표체적상관성。결과：DM환자위기층SMC초미결구명현이상，표현위봉극련접파배、포질내선립체종창、출현지갈소、세포막소요수량증가화세포막외기층증후。DM조위기층MHC、α-actin、calponin단백화mRNA표체수평균교대조조현저강저（P균﹤0.01），CML단백표체수평칙현저고우대조조（P﹤0.01）。DM조위기층MHC、calponin mRNA표체여CML단백표체정부상관（r=-0.59，P=0.02；r=-0.63，P=0.01），α-actin mRNA표체칙여CML단백표체무명현상관성（r=-0.49，P=0.06）。결론：DM환자존재위기층SMC초미결구파배，기층수축단백감소、AGEs증다차량자정부상관。상술변화가능영향SMC수축공능，진이삼여DM위동력장애적발생。
Background:Myopathy due to smooth muscle cells( SMC)abnormalities is involved in the pathogenesis of diabetic gastroparesis(DGP),however,the relationship between myopathy and advanced glycation end products(AGEs)is not fully clarified. Aims:To investigate the ultrastructural changes of gastric SMC in patients with diabetes mellitus( DM)and the relationship between expressions of contractile proteins and AGEs. Methods:Full-thickness gastric specimens from 30 gastric neoplasm patients undergoing gastrectomy from July 2012 to December 2012 at the First Affiliated Hospital with Nanjing Medical University were collected. Of them 15 patients had DM( DM group)and the other 15 patients without ( control group). Ultrastructure of SMC was observed by transmission electron microscopy. Expressions of three contractile proteins[ myosin heavy chains( MHC),α-actin and calponin]and Nε-carboxymethyllysine( CML),the key component of AGEs in gastric muscular layer were determined by Western blotting,and expressions of MHC,α-actin and calponin mRNA were determined by real-time PCR. Correlations of mRNA expressions of three contractile proteins with protein expression of CML were analyzed by Pearson correlation coefficient. Results:In DM group,significant ultrastructural changes were found in gastric SMC,including disruption of gap junction,swelling of mitochondria,occurring of lipofuscin in cytoplasm,increase in cell membrane alveolae,and thickening of basal lamina. Protein and mRNA expressions of muscular MHC,α-actin and calponin were significantly lower in DM group than in control group(P all ﹤0. 01),while protein expression of CML was significantly higher in DM group(P ﹤0. 01). In gastric muscular layer of DM group, negative correlations were found between expressions of MHC,calponin mRNA and CML protein(r= -0. 59,P=0. 02;r= -0. 63,P=0. 01),but no correlation was seen between α-actin mRNA and CML protein(r= -0. 49,P=0. 06). Conclusions:Disruption of SMC ultrastructure,decrease in contractile proteins and increase in AGEs are existed in gastric muscular layer of DM patients,and there is a negative correlation between contractile proteins and AGEs. These changes may cause impaired contractility of SMC,and subsequently lead to gastric motility disorders in diabetic patients.
