转录因子κB和早期生长反应因子1在急性肝内胆汁淤积性肝损伤大鼠模型中的表达

Expression of nuclear factor-κB and early growth response factor-1 in rat model with acute intrahepatic cholestatic hepatic injury

目的研究转录因子核因子κB(nuclear factor-κB,NF-κB)和早期生长反应因子1(early growthresponse factor-1,Egr-1)在急性肝内胆汁淤积性肝损伤大鼠模型中的表达及意义。方法将α-异硫氰酸萘酯(ANIT)按50 mg/kg 一次灌胃大鼠,建立急性肝内胆汁淤积性肝损伤的动物模型,免疫组织化学法和实时荧光定量 PCR 分别检测灌胃后24、48、72 h 肝组织中 NF-κB p65蛋白表达和 Egr-1 mRNA 表达量,全自动生化分析仪检测血清总胆红素(TB)、直接胆红素(DB)、丙氨酸氨基转移酶(ALT)、γ-谷氨酰转肽酶(GGT)。结果模型组24、48、72 h 肝组织 Egr-1 mRNA 表达量分别为(12.73±3.02)×10^(-2)、(8.19±2.29)×10^(-2)、(5.79±0.78)×10^(-2),24、48 h 均高于对照组(P 均<0.05),72 h 与对照组差异无统计学意义(P>0.05);模型组24、48、72 h肝组织 NF-κB p65核表达阳性细胞率分别为48.3%、26.5%、2.7%,其中24、48 h 均高于对照组(P 均<0.05),72 h 与对照组差异无统计学意义(P>0.05)。模型组24、48 h NF-κB p65核表达阳性细胞分级分别与同一时间点的血清 TB、DB、ALT 呈正相关(P 均<0.05);模型组24、48 hEgr-1 mRNA 表达量分别与同一时间点的血清TB、DB、AIT、GGT 呈正相关(P 均<0.05);模型组72 h 肝组织 NF-κB p65、Egr-1 mRNA 表达量与72 h 时血清 TB、DB、ALT、GGT 无相关关系(P 均>0.05);模型组24、48 h NF-κB p65核表达阳性细胞分级与同一时间点的血清 GGT 亦无相关关系(P 均>0.05);模型组24、48 h 肝组织 Egr-1 mRNA 表达量及 NF-κB p65核表达阳性细胞分级均与同一时间点肝组织病理损伤程度分级呈正相关(P 均<0.05);模型组72 h 肝组织 Egr-1 mR-NA 表达量及 NF-κB p65核表达阳性细胞分级均与同一时间点肝组织病理损伤程度分级无相关关系(P 均>0.05)。结论 NF-κB和 Egr-1在肝内胆汁淤积性肝损伤肝组织中表达增高,与肝损伤程度相关,参予了肝内胆汁淤积性肝损伤的病理过程。[临床儿科杂志,2008,26(10):879-884]

Objectives To explore the expression and significance of nuclear factor-κB （NF-κB） and early growth response factor-1 （Egr-1） in rat model with acute intrahepatic cholestatic hepatic injury. Methods A single dose （50 mg/kg） of α-naphthylisothiocyanate （ANIT） was administered by gavage to each experimental rat to induce intrahepatie cholestasis. The specimens of hepatic tissue and serum were collected from rats at 24 h, 48 h and 72 h after intoxication. The protein expression of NF-κB p65, the value of Egr-1 mRNA and serum TB, DB, ALT, GGT were assayed by immunohistochemistry, real-time PCR and fully-auto chemistry analyzer, respectively. Results In model group, at 24 h, 48 h and 72 h after intoxication, the positive rate of NF-κB p65 nuclear expression was48.3%, 26.5%, and 2.7%, respectively, which was higher than that of control at 24 h, 48 h （P 〈 0.05）, but was not significantly different at 72 h （P 〉 0.05）. In model group, at 24 h, 48 h and 72 h after intoxication, the value of Egr-1 rnRNA was （12.73 ±3.02） × 10^-2, （8.19 ± 2.29） ×10^-2 and （5.79 ± 0.78） × 10^-2, respectively, which was higher than that of control at 24 h and 48 h （P 〈 0.05）, but was not significantly different at 72 h （P 〈 0.05）. In model group, at 24 h and 48 h after intoxication, the grades of NF-κB p65 nuclear positive expression-cells was positively correlated to that of serum TB, DB, ALT （P 〈 0.05）, and the value of Egr-1 mRNA was positively correlated to that of serum TB, DB, ALT, GGT （P 〈 0.05）. In model group, at 72 h after intoxication, the grades of NF-KB p65 nuclear positive expression-cells and the value of Egr-1 mRNA were not correlated to that of serum TB, DB, ALT, GGT （P 〉 0.05）. In model group, at 24 h, 48 h after intoxication, the grades of NF-κB p65 nuclear positive expression-cells was not correlated to that of serum GGT （P 〉 0.05）. In model group, at 24 h, 48 h after intoxication, the value of Egr-1 mRNA and the grades of NF-KB p65 nuclear positive expression-cells were positively correlated to the degree of pathological injury in hepatic tissue （P 〈 0.05）. In model group at 72 h after intoxication, the value of Egr-1 mRNA and the grades of NF-KB p65 nuclear positive expression-cells were not correlated to the degree of pathological injury in hepatic tissue （P 〉 0.05）. Conclusions Acute intrahepatic cholestatic injury induced by ANIT resulted in the alteration of NF-κB protein expression and Egr-1 gene expression in hepatic tissue. The expression of NF-KB and Egr-1 was correlated to the degree of hepatic injury, which involves in pathological progress of intrahepatiecholestatic hepatic injury.