目的 观察大鼠腹膜内注射超氧歧化酶(SOD)抑制剂二乙基二硫代氨基甲酸盐(DETC)后胰腺的病理改变,并与大鼠胰管内注射三硝基苯磺酸(TNBS)所制备的慢性胰腺炎(CP)模型相比较.方法 将大鼠按随机表法分为DETC组、DETC对照组、TNBS组、TNBS对照组、正常对照组.DETC组大鼠腹膜内注射DETC 750 mg/kg体重,每周2次,DETC对照组在腹腔内注射等容积生理盐水.TNBS组大鼠胰管内注入含2％TNBS的乙醇磷酸盐缓冲液,TNBS对照组注入等容积乙醇磷酸盐缓冲液.正常对照组不做任何处理.术后2、4、6、8周分批处死大鼠,取血检测淀粉酶活性,取胰腺组织行病理及超微结构检查,检测组织内SOD、谷胱甘肽过氧化物酶(GSH-PX)活性和丙二醛(MDA)含量,免疫组化法检测组织α-平滑肌肌动蛋白(α-SMA),结蛋白(Desmin),胶原Ⅰ、Ⅲ,TGF-β1,纤维连接蛋白(FN)的表达,RT-PCR法检测组织TGF-β1 mRNA表达.结果 DETC组大鼠无死亡,TNBS组大鼠死亡率为15％.2组大鼠血淀粉酶活性差异无统计学意义.4周时DETC组大鼠胰腺纤维化评分为(3.4±1.1)分,显著高于TNBS组的(3.0±1.3)分(t=3.462,P＜0.05)；6周时胰腺组织腺体破坏评分为(9.1±1.8)分,显著高于TNBS组的(8.4±1.8)分(t=2.943,P＜0.05)；细胞空泡样变、脂肪浸润评分较TNBS组高,但差异均无统计学意义.DETC组和TNBS组在制模2周后即可见胰腺超微结构改变,4周后可见大量新生或已趋成熟的胶原纤维.2周时DETC组SOD活性较TNBS组显著下降(t=5.468,P＜0.05),GSH-PX活性在2、6周时较TNBS组显著下降,(t值分别为6.497,10.125,P＜0.05),而MDA活性在6、8周时较TNBS组均显著升高(t值分别为3.350,5.407,P值＜0.05).DETC组和TNBS组大鼠胰腺组织α-SMA,Desmin,胶原Ⅰ、Ⅲ,TGF-β1和FN表达及TGF-β1 mRNA表达水平差异均无统计学意义.结论 应用DETC持续抑制SOD活性可成功诱导CP.DETC组大鼠的胰腺脂肪浸润和纤维化程度较TNBS组出现得更早、更严重.采用该法制模操作简单,大鼠死亡率低,是一种较理想的制备CP模型的方法.
目的 觀察大鼠腹膜內註射超氧歧化酶(SOD)抑製劑二乙基二硫代氨基甲痠鹽(DETC)後胰腺的病理改變,併與大鼠胰管內註射三硝基苯磺痠(TNBS)所製備的慢性胰腺炎(CP)模型相比較.方法 將大鼠按隨機錶法分為DETC組、DETC對照組、TNBS組、TNBS對照組、正常對照組.DETC組大鼠腹膜內註射DETC 750 mg/kg體重,每週2次,DETC對照組在腹腔內註射等容積生理鹽水.TNBS組大鼠胰管內註入含2％TNBS的乙醇燐痠鹽緩遲液,TNBS對照組註入等容積乙醇燐痠鹽緩遲液.正常對照組不做任何處理.術後2、4、6、8週分批處死大鼠,取血檢測澱粉酶活性,取胰腺組織行病理及超微結構檢查,檢測組織內SOD、穀胱甘肽過氧化物酶(GSH-PX)活性和丙二醛(MDA)含量,免疫組化法檢測組織α-平滑肌肌動蛋白(α-SMA),結蛋白(Desmin),膠原Ⅰ、Ⅲ,TGF-β1,纖維連接蛋白(FN)的錶達,RT-PCR法檢測組織TGF-β1 mRNA錶達.結果 DETC組大鼠無死亡,TNBS組大鼠死亡率為15％.2組大鼠血澱粉酶活性差異無統計學意義.4週時DETC組大鼠胰腺纖維化評分為(3.4±1.1)分,顯著高于TNBS組的(3.0±1.3)分(t=3.462,P＜0.05)；6週時胰腺組織腺體破壞評分為(9.1±1.8)分,顯著高于TNBS組的(8.4±1.8)分(t=2.943,P＜0.05)；細胞空泡樣變、脂肪浸潤評分較TNBS組高,但差異均無統計學意義.DETC組和TNBS組在製模2週後即可見胰腺超微結構改變,4週後可見大量新生或已趨成熟的膠原纖維.2週時DETC組SOD活性較TNBS組顯著下降(t=5.468,P＜0.05),GSH-PX活性在2、6週時較TNBS組顯著下降,(t值分彆為6.497,10.125,P＜0.05),而MDA活性在6、8週時較TNBS組均顯著升高(t值分彆為3.350,5.407,P值＜0.05).DETC組和TNBS組大鼠胰腺組織α-SMA,Desmin,膠原Ⅰ、Ⅲ,TGF-β1和FN錶達及TGF-β1 mRNA錶達水平差異均無統計學意義.結論 應用DETC持續抑製SOD活性可成功誘導CP.DETC組大鼠的胰腺脂肪浸潤和纖維化程度較TNBS組齣現得更早、更嚴重.採用該法製模操作簡單,大鼠死亡率低,是一種較理想的製備CP模型的方法.
목적 관찰대서복막내주사초양기화매(SOD)억제제이을기이류대안기갑산염(DETC)후이선적병리개변,병여대서이관내주사삼초기분광산(TNBS)소제비적만성이선염(CP)모형상비교.방법 장대서안수궤표법분위DETC조、DETC대조조、TNBS조、TNBS대조조、정상대조조.DETC조대서복막내주사DETC 750 mg/kg체중,매주2차,DETC대조조재복강내주사등용적생리염수.TNBS조대서이관내주입함2％TNBS적을순린산염완충액,TNBS대조조주입등용적을순린산염완충액.정상대조조불주임하처리.술후2、4、6、8주분비처사대서,취혈검측정분매활성,취이선조직행병리급초미결구검사,검측조직내SOD、곡광감태과양화물매(GSH-PX)활성화병이철(MDA)함량,면역조화법검측조직α-평활기기동단백(α-SMA),결단백(Desmin),효원Ⅰ、Ⅲ,TGF-β1,섬유련접단백(FN)적표체,RT-PCR법검측조직TGF-β1 mRNA표체.결과 DETC조대서무사망,TNBS조대서사망솔위15％.2조대서혈정분매활성차이무통계학의의.4주시DETC조대서이선섬유화평분위(3.4±1.1)분,현저고우TNBS조적(3.0±1.3)분(t=3.462,P＜0.05)；6주시이선조직선체파배평분위(9.1±1.8)분,현저고우TNBS조적(8.4±1.8)분(t=2.943,P＜0.05)；세포공포양변、지방침윤평분교TNBS조고,단차이균무통계학의의.DETC조화TNBS조재제모2주후즉가견이선초미결구개변,4주후가견대량신생혹이추성숙적효원섬유.2주시DETC조SOD활성교TNBS조현저하강(t=5.468,P＜0.05),GSH-PX활성재2、6주시교TNBS조현저하강,(t치분별위6.497,10.125,P＜0.05),이MDA활성재6、8주시교TNBS조균현저승고(t치분별위3.350,5.407,P치＜0.05).DETC조화TNBS조대서이선조직α-SMA,Desmin,효원Ⅰ、Ⅲ,TGF-β1화FN표체급TGF-β1 mRNA표체수평차이균무통계학의의.결론 응용DETC지속억제SOD활성가성공유도CP.DETC조대서적이선지방침윤화섬유화정도교TNBS조출현득경조、경엄중.채용해법제모조작간단,대서사망솔저,시일충교이상적제비CP모형적방법.
Objective To investigate the pathologic changes in the pancreas of rats after intraperitoneal injection of DETC,a kind of superoxide dismutase (SOD) inhibitor,and to compared that with another model of chronic pancreatitis by pancreatic duct injection of TNBS.Methods The rats were randomly divided into DETC group,DETC control group,TNBS group,TNBS control group,normal control group.Rats in DETC group received an intra-peritoneal injection of DETC twice a week,and rats in DETC control group received an intra-peritoneal injection of same amount of normal saline.Rats in TNBS group was injected with 2％ TNBS ethanol phosphate buffer into the pancreatic duct,while rats in TNBS control group was treated with injection of same amount of ethanol phosphate buffer,and rats in normal control group received no treatment.The rats were sacrificed after 2 w,4 w,6 w and 8 w.The serum levels of amylase were determined,and pathological and ultrastructure changes of the pancreas were measured.The levels of SOD,GSH-PX activity and MDA content were detected.The expressions of α-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN in tissue were detected by immunohistochemical assay.The TGF-β1 mRNA expression was detected by RTPCR.Results No rat died in DETC group.The mortality rate of TNBS group was 15％.The serum levels of amylase were not statistically different between the 2 groups.The fibrosis scores of rat in DETC group at 4 w was 3.4 ± 1.l,which was significantly higher than that in TNBS group (3.0 ± 1.3,t =3.462,P ＜ 0.05).At 6 w,the damage scores of rat in DETC group was 9.1 ± 1.8,which was significantly higher than that in TNBS group (8.4 ± 1.8,t =2.943,P ＜ 0.05).Scores of vacuolar degeneration and fatty infiltration of rat in DETC group were higher than those in TNBS group,but the difference between the two groups was not statistically significant.Two weeks later,ultrastructure changes of pancreas could be observed,and large amounts of regenerative or mature collagen could be seen at 4 w.The SOD activity of DETC group was significantly decreased when compared with those in TNBS group (t =5.468,P ＜ 0.01).The GSH-PX activity of DETC group at 2 w,6w was significantly decreased when compared with those in TNBS group (t =6.497,10.125,P＜0.01).While the activity of MDA at 6 w,8 w was significantly increased when compared with those in TNBS group (t =3.350,5.407,P ＜0.05).The differences at other time points were not statistically significant.The expressions of (a)-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN,and TGF-β1 mRNA were not statistically significant between the 2 groups.Conclusions Sustained suppression of SOD activity can successfully induce chronic pancreatitis.Fatty infiltration and fibrosis in pancreas in DETC group occurs earlier with more severe presentation than that in TNBS group.Intraperitoneal injection of DETC is easy with low mortality rate,which is an ideal method for chronic pancreatitis model induction.
