应用c-myc shRNA治疗慢性增生性胆管炎在肝内胆管结石治疗中的价值  

Value of Treatment for Hepatolithiasis by Using c-myc shRNA in Chronic Proliferative Cholangitis

作　　者：周勇[1] 李富宇[1] 王晓东[2] 李宁[1] 程南生[1] 蒋力生[1] 何生[1] 

机构地区：[1]四川大学华西医院肝胆外科,成都610041 [2]佳木斯市中心医院普外科,黑龙江佳木斯154001

出　　处：《中国普外基础与临床杂志》2010年第10期1039-1045,共7页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金(项目编号:30801111);教育部博士点新教师基金(项目编号:200806101065);四川省科技支撑计划(项目编号:10ZC1705)~~

摘　　要：目的探讨能否通过局部应用c-myc shRNA抑制慢性增生性胆管炎(CPC)增殖相关基因的表达而抑制CPC的过度增殖行为和成石潜力。方法经十二指肠乳头向胆总管逆行插入5-0尼龙缝合线建立CPC动物模型(CPC组)。c-myc shRNA治疗组在CPC组基础上向胆总管内分别注入3种c-myc shRNA即分别为c-myc shRNA-1、c-myc shRNA-2及c-myc shRNA-3,另设阴性对照组和假手术组。应用HE、Massion和PAS/AB染色观察胆管组织病理学变化,免疫组化法检测c-myc蛋白表达,免疫荧光法检测5-溴脱氧尿核苷(BrdU)蛋白表达,实时(RT)-PCR检测c-myc、Mucin 3和ProcollagenⅠmRNA的表达,Western blot法检测Ki-67蛋白的表达,改良Fisherman法检测β-葡萄糖醛酸酶(β-G)的活性。结果①与CPC组及阴性对照组比较,c-myc shRNA治疗组的胆管黏膜上皮(HE染色)、黏膜下酸性黏液腺体(PAS/AB染色中蓝色)和胆管壁胶原纤维(Massion染色蓝染)的过度增生程度明显减弱,BrdU蛋白表达也明显减弱。②c-myc、Mucin 3和ProcollagenⅠmRNA,c-myc蛋白,Ki-67蛋白,以及β-G活性在c-myc shRNA治疗组均明显低于CPC组及阴性对照组(P<0.05),但均高于假手术组(P<0.05)。结论 c-myc shRNA治疗能够有效地抑制CPC的过度增殖行为和成石潜力,可能会有助于预防胆管再狭窄和肝内胆管结石的术后复发。Objective To determine whether local delivery of c-myc shRNA could inhibit hyperplasia and lithogenic potentiality in a rat model of chronic proliferative cholangitis（CPC） via specific blockade of the c-myc expression.Methods The CPC animal model（CPC group） was established via retrograde insertion of a 5-0 nylon thread into the common bile duct through Vater＇s papilla.Three kinds of c-myc shRNAs were then respectively injected in c-myc shRNA group,which were included shRNA-1,shRNA-2,and shRNA-3,respectively.Negative control group and sham operation group were established for comparison.Subsequently,histopathological changes of bile duct wall were observed by HE,Massion,and PAS/AB staining;c-myc protein was detected by immunohistochemistry method;5-bromodeoxyuridine（BrdU） protein was tested by immumofluorescence method;c-myc,Mucin 3,and Procollagen Ⅰ mRNAs were detected by real time PCR;Ki-67 protein was determined by Western blot;Activity of β-glucuronidase was measured by modified Fisherman method.Results ①Compared with the CPC and negative control groups,biliary tract mucosa epithelium（HE staining）,submucosal acid mucinous gland（mid-blue staining,PAS/AB staining）,and degree of over-hyperplasia of collagen fiber in bile duct wall（blue staining,Massion staining） were weaker in the c-myc shRNA group.②The expressions of c-myc mRNA,Mucin 3 mRNA,Procollagen Ⅰ mRNA,Ki-67 protein,and β-G activity in the c-myc shRNA group were lower than those of the CPC and negative control groups（P0.05）,but higher than those of the sham operation group（P0.05）.Conclusion c-myc shRNA treatment could effectively inhibit the hyperplastic behavior and lithogenic potential of CPC,which might help to prevent the biliary restenosis and stone recurrence.

关 键 词：肝内胆管结石 慢性增生性胆管炎 复发 胆管狭窄 c-mycshRNA 

分 类 号：R657.4[医药卫生—外科学]