表皮生长因子受体拮抗剂对慢性增生性胆管炎的实验疗效探讨

Experimental Study of Epidermal Growth Factor Receptor Antagonist in Treating Chronic Proliferative Cholangitis

目的探讨表皮生长因子受体(EGFR)拮抗剂能否抑制慢性增生性胆管炎(CPC)的过度增殖行为和成石潜力,以期为肝内胆管结石的防治探索新的治疗途径。方法将46只健康SD大鼠随机分为5组:CPC模型组(10只),仅行造模处理;3种剂量EGFR拮抗剂AG-1478治疗组(分为3mg/kg、6mg/kg和12mg/kg组,各10只),在行造模术的同时经胆总管内注入相应剂量的AG-1478,术后7d内腹腔内注射AG-1478 1.5mg/(kg.d);假手术组(SO组,6只),仅行开腹术后即关腹。于术后7d剖腹取胆管标本组织分别行病理组织学观察、免疫组化染色、RT-PCR和Western blot检测,比较各组EGFR、5-溴脱氧尿核苷(BrdU)、Ki-67、黏蛋白5AC和Ⅰ型胶原蛋白的mRNA或蛋白表达情况,以评估AG-1478对病变胆管黏膜过度增殖行为(EGFR、Ki-67、BrdU、Ⅰ型胶原蛋白)和成石潜力(黏蛋白5AC)的影响。结果 与CPC模型组比较,治疗组的EGFR、Ki-67及BrdU表达明显降低,组织学观察所见的胆管黏膜上皮和胶原纤维的过度增殖行为也得到了有效的抑制;此外,治疗组的黏蛋白5AC mRNA和Ⅰ型胶原蛋白表达水平也较CPC模型组明显降低(P<0.05)。结论 EGFR拮抗剂可有效抑制CPC病变胆管黏膜的过度增殖和成石潜力,有望成为CPC新的治疗手段。

Objective To investigate the effectiveness of epidermal growth factor receptor antagonist（AG-1478） on chronic proliferative cholangitis（CPC）,so as to investigate new treatment approach for hepatolithiasis associated with CPC.Methods Forty-six SD rats were divided into 5 groups： CPC model group（n=10）,only made models.AG-1478 treatment group（divided into 3 mg/kg,6 mg/kg,and 12 mg/kg groups,n=10 per group）,the common bile ducts in CPC animal model received an intralumenal administration of AG-1478 at the meantime of modeling,followed by intraperitoneal AG-1478 injection of 1.5 mg/（kg·d） for 7 days.Sham operation group（SO group,n=6）.Subsequently,histopathological observation,immunohistochemistry,real time PCR,and Western blot were used to evaluate the mRNA expression and influence of AG-1478 on the hyperplasia（EGFR,ki-67,BrdU,collagen Ⅰ protein） and lithogenic potential（Mucin 5AC） of CPC.Results Compared with CPC model group,the expressions of EGFR,ki-67,and BrdU were obviously decreased in the AG-1478 treatment group.Also,the inhibition of hyperplasia of biliary epithelium and collagen fibers were confirmed by histopathological observation.Additionally,the expressions of Mucin 5AC mRNA and collagen Ⅰ protein remarkable decreased in the AG-1478 treatment group（P0.05）.Conclusions EGFR inhibitor（AG-1478） could shows inhibitory effectivenss on the CPC-mediated hyperplasia and lithogenic potential,and therefore holds promise as the new treatment approach for CPC.