沉默纤维介素2基因表达对急性坏死性胰腺炎小鼠的治疗作用  

Therapeutic effects of silencing fibrinogen-like protein 2 gene on acute necrotizing pancreatitis mice

作　　者：邵利梅 叶晓华[1] 丁进[1] Shao Limei;Ye Xiaohua;Ding Jin(Department of Gastroenterology,Jinhua Hospital of Zhejiang University,Jinhua 321000,China)

机构地区：[1]浙江省金华市中心医院,浙江金华321000

出　　处：《中华胰腺病杂志》2018年第5期328-332,共5页Chinese Journal of Pancreatology

基　　金：浙江省科技厅公益项目（2015C37120）;金华市科技局社会发展一般项目（2014-3-053）

摘　　要：目的探讨沉默纤维介素2（FGL2）基因表达对急性坏死性胰腺炎（ANP）小鼠的治疗作用。方法18只C57/BL小鼠按数字表法随机分为对照组、ANP组、携带靶向FGL2的siRNA腺病毒（Ad-FGL2-siRNA）组，每组6只。应用胰胆管逆行注射牛磺胆酸钠法诱导ANP模型，Ad-FGL2-siRNA组在诱导ANP前尾静脉注射0.5 ml 4×108 pfu/ml的Ad-FGL2-siRNA。6 h后处死小鼠，收集胰腺组织及血液标本。胰腺组织行常规病理学检查，采用ELISA法检测血清TNF-α和IL-1β水平，RT-PCR、蛋白质印迹法及免疫组织化学染色法检测胰腺组织FGL2mRNA及蛋白表达，TUNEL法检测细胞凋亡。结果对照组、ANP组、Ad-FGL2-siRNA组小鼠胰腺组织的病理评分分别为（1.33±0.21）、（9.17±0.98）、（6.26±0.52）分；血清TNF-α水平分别为（63.8±4.2）、（240.4±18.6）、（123.0±10.3）ng/L；IL-1β水平分别为（43.6±4.4）、（186.6±18.7）、（92.0±10.9）ng/L；胰腺组织FGL2 mRNA相对表达量分别为1.20±0.22、4.40±1.21、2.15±0.56；FGL2蛋白相对表达量分别为0.33±0.08、1.23±0.24、0.68±0.09；FGL2阳性细胞率为（2.56±0.31）%、（15.10±3.23）%、（8.68±1.81）%；细胞凋亡数为（4.51±1.21）、（35.81±4.11）、（11.79±3.02）个/200倍视野。ANP组较对照组均显著升高，Ad-FGL2-siRNA组较ANP组均显著下降，除FGL2 mRNA表达量外Ad-FGL2-siRNA组均显著高于对照组，差异均有统计学意义（P值均〈0.05）。结论沉默FGL2基因表达可以减少炎症因子释放、抑制细胞凋亡从而减轻ANP小鼠胰腺组织损伤。Objective To investigate the therapeutic effect of silencing fibrinogen-like protein 2（FGL2） gene on acute necrotic pancreatitis （ANP） mice.Methods Eighteen C57/BL mice were randomly divided into SO （sham operation） group, ANP group and Ad-FGL2-siRNA group carrying FGL2 siRNA adenovirus, with 6 in each group. Sodium taurocholate was retrogradely injected into the biliopancreatic ducts of the mice to induce ANP mice model. The mice in Ad-FGL2-siRNA group were injected intravenously in the tail vein with Ad-FGL2-siRNA befor the model establishment. The mice were sacrificed 6 h later, and then the pancreatic tissue and blood were collected. TNF-α and IL-1β expression were measured with ELISA, pancreatic tissue was examined with routine pathological examination, FGL2 mRNA and protein expression were measured with reverse transcription-PCR, western blotting and immunohistochemical staining, and cell apoptosis was assessed by TUNEL method.Results The pathological score of the pancreatic tissue in SO group, ANP group and Ad-FGL2-miRNA group was（1.33±0.21）, （9.17±0.98） and （6.26±0.52）, respectively. The serum TNF-α level in SO group, ANP group and Ad-FGL2-miRNA group was（63.8±4.2）, （240.4±18.6） and（123.0±10.3）ng/L, respectively. The serum IL-1β level was （43.6±4.4）, （186.6±18.7）and （92±10.9）ng/L. The mRNA expressions of FGL2 was 1.20±0.22, 4.40±1.21 and 2.15±0.56. The protein expressions of FGL2 was 0.33±0.08, 1.23±0.24 and 0.68±0.09. The rate of FGL2 positive cells was（2.56±0.31）%, （15.10±3.23）% and（8.68±1.81）%. The number of apoptotic cells was（4.51±1.21）, （35.81±4.11） and（11.79±3.02）/×200HPF, which in the ANP group was higher than that in SO group, and in the Ad-FGL2-siRNA group was significantly lower than that in ANP group and higher than that in SO group. All the differences were statistically significant （all P values 〈0.05）, except that on the FGL2 mRNA expression between Ad-FGL2-siRNA group and SO group.Conclu

关 键 词：胰腺炎 急性坏死性 纤维蛋白原 纤维介素2 细胞凋亡 炎症因子 

分 类 号：R576[医药卫生—消化系统]