建立大鼠胆管良性狭窄模型的实验研究
摘要
目的：
探讨建立简捷、经济实用、稳定可靠的大鼠胆管良性狭窄动物模型的方法,利于对胆管良性狭窄的发生机制和有效治疗方法的研究。
方法：
49只雌性SD (Sprague Dawley)大鼠体重220-260克,平均240±10 g,随机将其分为实验组21只、实验对照组21只和空白对照组7只,所有实验动物均采用3%的戊巴比妥注射液进行腹腔麻醉。实验组21只动物进行腹腔麻醉后上腹剑突下正中切口开腹,显露胆总管后在距离十二指肠上缘2cm处分离出胆总管长约1cm,其下面放置一厚约0.2cm的棉垫后滴注95%的酒精约0.7ml于胆管上持续2-3分钟后关腹；实验对照组以同样的方法进行手术,在胆管上滴注0.9%的生理盐水后2-3分钟关腹；空白对照组只是找出胆管后就关腹,不进行其他处理。术后观察包括精神状态、尿液颜色、进食情况;实验模型组和实验对照组术后7、14、21天共分三次每次分别取7只动物称体重与及时抽血检查肝功能(ALT、GGT、T-BIL、D-BIL),并切取部分肝脏和实验处胆管行病理学检查,21天组还进行胆道造影了解胆管梗阻情况。统计学采用SPSS软件t检验及多元方差分析。
结果：
术后1hSD大鼠能翻身活动,3-4h后自主饮水,逐渐恢复饮食。①从术前到术后21天,实验造模组检测血清丙氨酸氨基转移酶(ALT)、γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)、血清总胆红素(T-BIL)、血清直接胆红素(D-BIL)等指标有明显变化(P<0.05)；而对照组和空白对照组无显著变化。②各时间点处死大鼠切取肝脏及胆管行病理学检查,实验造模组随着时间推移发现肝细胞有不同程度损伤,胆管组织部分纤维增生；而对照组和空白对照组术前与术后对比无明显变化。③术后21天胆道造影发现,实验造模组胆管有狭窄表现；而对照组和空白对照组无胆管狭窄出现。通过上述比较,实验组与对照组差异有统计学意义(P<0.05),对照组与空白对照组无统计学意义(P>0.05)。
结论：
通过肝外胆管外滴注95%的乙醇制造大鼠胆管良性狭窄模型是可行的,能够为胆管良性狭窄的研究提供另一种动物模型。
Objective
Explore a simple、economical、practical stable and reliable rat model of bile duct benign stricture, to research the pathogenesis and effective treatment methods of bile duct benign stricture.
Methods
49female SD rats, weight 220-260 g, average 240±10 g, randomly divided into the experimental group 21、the control group 21 and the blank control group7,all the experimental animals will be injected with the 3% of pentobarbital for celiac anesthesia. the experimental group was laparotomy under sword axon in epigastric abdominal next midline incision after anesthesia,exposue the common bile duct and separation it on 1 cm in the distance from Duodenum, placed a gauze thick about 0.2cm behind the common bile duct, dripped about 0.7ml of the 95% alcohol on it to continue 2-3minutes and then closed abdominal; The control group was surgeryed as the same method except for dripping the Physiological saline of 0.9%. The blank control group only search for the bile duct without doing other thing.We must observe the mental state、urine color、eating situation after the surgery and so on. Selected 5 tested animal to weigh and draw blood to test the liver function(include ALT、GGT、T-BIL、D-BIL),and cut a part liver and the tested bile duct for pathology examination after the postopration 7、14、21 day, in order to understand the situation of bile duct obstruction through biliary imaging after the postopration 21 day. Using SPSS software, t -test and multivariate analysis of variance.
Results
SD rats turn over in 1hour, water after 3-4h, gradual return food.①the experimental group test the ALT、GGT、T-BIL、D-BIL etc.There is a obvious change(P<0.05)from preoperation to the 21 day of postoperation;But the control group no obvious change.②Killed the rat in the right time to cut the liver and bile duct for pathology examination, to find liver cell vary degrees injury and the part of the bile duct organization fiber hyperplasia in the experiment group,however there is no variation on the control group from preopration to postopration.③There is a obvious bile duct obstruction through biliary imaging after the 21 day of postopration.but the control group no bile duct obstruction. Through the comparison, the experimental group and the control group was statistically significant (P<0.05), the control group and the blank control group was not statistically significant (P> 0.05).
Conclusions
It is viable to drip the 95% alcohol on the outside of liver bile duct to build the bile duct benign stricture on rat in this experiment,it can offer another good animal model for research the bile duct benign stricture.
探讨建立简捷、经济实用、稳定可靠的大鼠胆管良性狭窄动物模型的方法,利于对胆管良性狭窄的发生机制和有效治疗方法的研究。
方法：
49只雌性SD (Sprague Dawley)大鼠体重220-260克,平均240±10 g,随机将其分为实验组21只、实验对照组21只和空白对照组7只,所有实验动物均采用3%的戊巴比妥注射液进行腹腔麻醉。实验组21只动物进行腹腔麻醉后上腹剑突下正中切口开腹,显露胆总管后在距离十二指肠上缘2cm处分离出胆总管长约1cm,其下面放置一厚约0.2cm的棉垫后滴注95%的酒精约0.7ml于胆管上持续2-3分钟后关腹；实验对照组以同样的方法进行手术,在胆管上滴注0.9%的生理盐水后2-3分钟关腹；空白对照组只是找出胆管后就关腹,不进行其他处理。术后观察包括精神状态、尿液颜色、进食情况;实验模型组和实验对照组术后7、14、21天共分三次每次分别取7只动物称体重与及时抽血检查肝功能(ALT、GGT、T-BIL、D-BIL),并切取部分肝脏和实验处胆管行病理学检查,21天组还进行胆道造影了解胆管梗阻情况。统计学采用SPSS软件t检验及多元方差分析。
结果：
术后1hSD大鼠能翻身活动,3-4h后自主饮水,逐渐恢复饮食。①从术前到术后21天,实验造模组检测血清丙氨酸氨基转移酶(ALT)、γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)、血清总胆红素(T-BIL)、血清直接胆红素(D-BIL)等指标有明显变化(P<0.05)；而对照组和空白对照组无显著变化。②各时间点处死大鼠切取肝脏及胆管行病理学检查,实验造模组随着时间推移发现肝细胞有不同程度损伤,胆管组织部分纤维增生；而对照组和空白对照组术前与术后对比无明显变化。③术后21天胆道造影发现,实验造模组胆管有狭窄表现；而对照组和空白对照组无胆管狭窄出现。通过上述比较,实验组与对照组差异有统计学意义(P<0.05),对照组与空白对照组无统计学意义(P>0.05)。
结论：
通过肝外胆管外滴注95%的乙醇制造大鼠胆管良性狭窄模型是可行的,能够为胆管良性狭窄的研究提供另一种动物模型。
Objective
Explore a simple、economical、practical stable and reliable rat model of bile duct benign stricture, to research the pathogenesis and effective treatment methods of bile duct benign stricture.
Methods
49female SD rats, weight 220-260 g, average 240±10 g, randomly divided into the experimental group 21、the control group 21 and the blank control group7,all the experimental animals will be injected with the 3% of pentobarbital for celiac anesthesia. the experimental group was laparotomy under sword axon in epigastric abdominal next midline incision after anesthesia,exposue the common bile duct and separation it on 1 cm in the distance from Duodenum, placed a gauze thick about 0.2cm behind the common bile duct, dripped about 0.7ml of the 95% alcohol on it to continue 2-3minutes and then closed abdominal; The control group was surgeryed as the same method except for dripping the Physiological saline of 0.9%. The blank control group only search for the bile duct without doing other thing.We must observe the mental state、urine color、eating situation after the surgery and so on. Selected 5 tested animal to weigh and draw blood to test the liver function(include ALT、GGT、T-BIL、D-BIL),and cut a part liver and the tested bile duct for pathology examination after the postopration 7、14、21 day, in order to understand the situation of bile duct obstruction through biliary imaging after the postopration 21 day. Using SPSS software, t -test and multivariate analysis of variance.
Results
SD rats turn over in 1hour, water after 3-4h, gradual return food.①the experimental group test the ALT、GGT、T-BIL、D-BIL etc.There is a obvious change(P<0.05)from preoperation to the 21 day of postoperation;But the control group no obvious change.②Killed the rat in the right time to cut the liver and bile duct for pathology examination, to find liver cell vary degrees injury and the part of the bile duct organization fiber hyperplasia in the experiment group,however there is no variation on the control group from preopration to postopration.③There is a obvious bile duct obstruction through biliary imaging after the 21 day of postopration.but the control group no bile duct obstruction. Through the comparison, the experimental group and the control group was statistically significant (P<0.05), the control group and the blank control group was not statistically significant (P> 0.05).
Conclusions
It is viable to drip the 95% alcohol on the outside of liver bile duct to build the bile duct benign stricture on rat in this experiment,it can offer another good animal model for research the bile duct benign stricture.
引文
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[1]Xu J, Geng zM, Ma QY.Microstructural and ultrastructural chang in the healing Process of bile duct trauma. Hepatobiliary Pancreat Dis Int,2003;2:295-299.
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