摘要
目的观察核因子(NF)-κB、细胞间黏附分子(ICAM)-1和炎症细胞因子在急性胰腺炎(AP)肝损伤中的作用。方法36只SD大鼠随机分为AP组、假手术组(SO组),通过胰胆管逆行注射3.5%牛磺胆酸钠溶液制作大鼠AP模型。术后3、6、12 h胰腺组织光镜检查;肝组织光镜、电镜观察;检测血清丙氨酸氨基转移酶(ALT);放射免疫法(RIA)测定血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6;酶联免疫吸附法(ELISA)检测IL-10;实时定量聚合酶链反应(Real-time PCR)检测肝组织TNF-αmRNA、IL-6 mRNA;肝组织Envision法免疫组织化学测定NF-κB、ICAM-1。结果病理学证实建立AP组,出现肝损伤的病理形态变化;与SO组比较,AP组的血清ALT以及TNF-α、IL-6、IL-1β、IL-10水平显著增高(P<0.05),其中血TNF-α水平在3 h升高、6 h达高水平,血IL-6在AP 3 h明显升高,下降缓慢。AP 6、12 h组肝NF-κB活化,AP各组间肝NF-κB活性表达差异均有统计学意义(x^2=15.048,P<0.05),ICAM-1无表达。AP 3～12 h内,AP组肝TNF-αmRNA在3 h明显升高、6 h高表达;AP组的肝IL-6 mRNA则在3 h已增加、3 h后缓慢下降,均显著高于各自时段的SO组(P<0.05)。结论AP发生后,肝脏NF-κB活化,可能介导肝TNF-αmRNA、IL-6 mRNA的表达来参与肝损伤,ICAM-1对AP早期肝损伤元明显作用。
Objective To investigate the role of nuclear-κB （NF-κB）, intercellular adhesion molecule-1 （ ICAM-1 ） and inflammatory cytokines in acute pancreatitis （AP） with liver injury in rats. Methods Thirty-six SD rats were randomly allocated into acute pancreatitis group （AP group） and sham operation group （ SO group）. AP model was induced by retrograde injection of 3.5% sodium taurocholate into the pancreatic duct in rats. The histopathological changes of the pancreas were observed and studied by light microscopy at 3,6 and 12 h after operation. The pathological changs of liver were examined under light microscopy and electron microscopy. The serum levels of alanine aminotransferase （ALT） were determined,the changes in blood levels of TNF-α, IL-1β and IL-6 were tested by radioimmunoassay （RIA） method,and the levels of serum IL-10 were measured by enzyme linked immunosorbent assay （ELISA） at different time points. The expression of TNF-α and IL-6 mRNA in the liver was detected by real time polymerase chain reaction （ RT-PCR）. NF-κB and ICAM-1 activity in the liver was examined by immunohistochemical methods. Results Histopathologic examination revealed the occurrence of pancreatic inflammation in AP group. Pathological changes appeared in liver of AP group. With the development of AP, the levels of serum ALT, TNF-α, IL-1β, IL-6 and IL-10 in AP group were increased significantly as compared with SO group （ P 〈 0.05 ） ： the serum levels of TNF-α were increased significantly at 3-6 h, and reached peak at 6 h;the serum levels of IL-6 were increased significantly at 3 h and then decreased sligntly. NF-κB activity was strengthened at 6 and 12 h,but there was no change in ICAM-1. There was significant difference in the NF-κB expression among the three AP groups （χ^2 = 15. 048, P 〈 0.05）. The expression of TNF-α and IL-6 mRNA in liver tissue of the AP group was also stronger than that in SO group in 3-12 h （ P 〈 0.05 ） , and the expression of TNF-α mRNA was increased at 3 h and reached the peak at 6 h. The expression of IL-6 mRNA reached the peak at 3 h and was only decreased sligntly after 3 h. Conclusion The activation of hepatic NF-κB is associated with the liver injury by regulating the expression of TNF-α and IL-6 mRNA in AP. ICAM-1 has no obvious effect on the liver injury in the early stage of AP.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第9期1083-1086,共4页
Chinese Journal of Experimental Surgery
