内毒素血症肝损伤幼年大鼠肝脏NF-κB的变化

胡泽华; 王琳琳; 唐清; 单庆文; 连淑君; 陈萍; 陈秀奇

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中国当代儿科杂志 ›› 2010, Vol. 12 ›› Issue (10) : 804-808.

论著·实验研究

内毒素血症肝损伤幼年大鼠肝脏NF-κB的变化

胡泽华，王琳琳，唐清，单庆文，连淑君，陈萍，陈秀奇

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NF-κB levels in the liver of young rats with endotoxemic liver injury

HU Ze-Hua, WANG Lin-Lin, TANG Qing, SHAN Qing-Wen, LIAN Shu-Jun, CHEN Ping, CHEN Xiu-Qi

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摘要

目的：研究内毒素血症肝损伤幼年大鼠肝脏NF-κB的变化及与TNF-α、IL-6的关系。方法：18日龄Wistar大鼠40只随机分为正常对照组和内毒素血症组，内毒素血症组根据制模后取标本时间不同分2 h、6 h、12 h、24 h 4个亚组（n=8）。内毒素血症组腹腔注射LPS 5 mg/kg制备内毒素血症动物模型。观察各组肝细胞的病理改变，ELISA测定肝组织匀浆TNF-α、IL-6含量，赖氏法测血清丙氨酸转氨酶（ALT）浓度，免疫组化法检测肝组织NF-κB活化水平。结果：光镜下内毒素血症组肝组织损伤以6 h最明显，损伤等级评分在各个时间点均较正常对照组高（P<0.05）；内毒素血症组ALT水平6 h、12 h及24 h均较正常对照组增高（P<0.05）；内毒素血症组各个时间点肝细胞NF-κB p65蛋白表达的阳性细胞百分比均较正常对照组高（P<0.05）；肝组织TNF-α、IL-6检测显示内毒素血症组6 h、12 h高于正常对照组（P<0.05）。结论：内毒素血症可引起肝损伤，导致肝细胞的破坏和肝功能的改变；NF-κB的活化参与了内毒素血症肝损伤的发生，可能是通过介导炎症因子TNF-α和IL-6的合成使肝细胞发生炎性损伤。［中国当代儿科杂志，2010，12（10）：804-808］

Abstract

OBJECTIVE: To study hepatic NF-κB level following endotoxemic liver injury, and its relationship with hepatic TNF-α and IL-6 levels in young rats. METHODS: Forty 18-day-old rats were randomly assigned to a normal control and an endotoxemia group. Endotoxemia was induced by lipopolysaccharide injection (LPS, 5 mg/kg). The endotoxemia group was subdivided into four groups sampled at 2, 6, 12 and 24 hrs after LPS injection (n=8 each). Pathological changes in liver cells were observed under a light microscope. TNF-α and IL-6 levels in liver tissue homogenates were measured using ELISA. Reitman-Frankel was used to measure serum ALT concentrations. NF-κB activation level in liver tissues was detected by immunohistochemistry. RESULTS: Liver tissue injury was the most obvious 6 hrs after LPS injection under the light microscope, and the damage rating of liver tissues was significantly higher in the endotoxemia group than that in the normal control group at all time points (P<0.05). ALT levels in the endotoxemia group were significantly higher than those in the normal control group 6, 12 and 24 hrs after LPS injection (P<0.05). NF-κB p65 protein expression in liver cells (percentage of nuclear positive cells) in the endotoxemia groups was significantly higher than that in the normal control group (P<0.05). TNF-α and IL-6 levels in liver tissue homogenates in the endotoxemia groups were significantly higher than those in the normal control group 6 and 12 hrs after LPS injection (P<0.05). CONCLUSIONS: Endotoxemia can cause liver injury, resulting in liver cell damage and changes in liver function. NF-κB activation is involved in endotoxemic liver injury which may be mediated by inflammatory cytokines TNF-α and IL-6 synthesis. ［Chin J Contemp Pediatr, 2010, 12 (10):804-808］

导出引用

胡泽华, 王琳琳, 唐清, 单庆文, 连淑君, 陈萍, 陈秀奇. 内毒素血症肝损伤幼年大鼠肝脏NF-κB的变化[J]. 中国当代儿科杂志. 2010, 12(10): 804-808

HU Ze-Hua, WANG Lin-Lin, TANG Qing, SHAN Qing-Wen, LIAN Shu-Jun, CHEN Ping, CHEN Xiu-Qi. NF-κB levels in the liver of young rats with endotoxemic liver injury[J]. Chinese Journal of Contemporary Pediatrics. 2010, 12(10): 804-808

中图分类号： R-33

参考文献

［1］Xu FL, You HB, Li XH, Chen XF, Liu ZJ, Gong JP. Glycine attenuates endotoxin-induced liver injury by downregulating TLR4 signaling in Kupffer cells［J］. Am J Surg, 2008, 196(1): 139-148.
［2］Zhang D, Jiang J, Jiang S, Ma J, Su T, Qiu L, et al. Molecular characterization and expression analysis of a putative LPS-induced TNF-alpha factor (LITAF) from pearl oyster Pinctada fucata［J］. Fish Shellfish Immunol, 2009, 27(3): 391-396.
［3］谢国旗,蒋建新,陈永华,刘大威,朱佩芳,王正国.内毒素致急性肝损伤机制的实验研究［J］.中华急诊医学杂志,2002,11(1):6-9.
［4］Camargo CA, Madden JF, Gao W, Selvan RS, Clavien PA. Interleukin-6 protects liver against warm ischemia/reperfusion injury and promotes hepatocyte proliferation in the rodent［J］. Hepatology, 1997, 26(6): 1513-1520.
［5］Wang YY, Dahle MK, Steffensen KR, Reinholt FP, Collins JL, Thiemermann C, et al. Liver X receptor agonist GW3965 dose-dependently regulates lps-mediated liver injury and modulates posttranscriptional TNF-alpha production and p38 mitogen-activated protein kinase activation in liver macrophages［J］. Shock, 2009, 32(5): 548-553.
［6］Thomson RK, Arthur MJ. Mechanisms of liver cell damage and repair［J］. Eur J Gastroenterol Hepatol, 1999, 11(9): 949-955.
［7］Shibayama Y, Asaka S, Nishijima A, Nakata K. A study of endotoxin-associated hepatotoxicity on proliferating hepatocytes［J］. Exp Mol Pathol, 1992, 56(1): 70-75.
［8］Ishii K, Ito Y, Katagiri H, Matsumoto Y, Kakita A, Majima M. Neutrophil elastase inhibitor attenuates lipopolysaccharide-induced hepatic microvascular dysfunction in mice［J］. Shock, 2002, 18(2): 163-168.
［9］Qiu Z, Kwon AH, Tsuji K, Kamiyama Y, Okumura T, Hirao Y. Fibronectin prevents D-galactosamine /lipopolysaccharide-induced lethal hepatic failure in mice［J］. Shock, 2006, 25(1): 80-87.
［10］Dhainaut JF, Marin N, Mignon A, Vinsonneau C. Hepatic response to sepsis: interaction between coagulation and inflammatory［J］. Crit Care Med, 2001, 29(7 Suppl): S42-47.
［11］Amacher DE. A toxicologist′s guide to biomarkers of hepatic response［J］. Hum Exp Toxicol, 2002, 21(5): 253-262.
［12］Ozer J, Ratner M, Shaw M, Bailey W, Schomaker S. The current state of serum biomarkers of hepatotoxicity［J］. Toxicology, 2008, 245(3): 194-205.
［13］Monsalve E, Ruiz-García A, Baladrón V, Ruiz-Hidalgo MJ, Sánchez-Solana B, Rivero S, et al. Notch1 upregulates LPS-induced macrophage activation by increasing NF-kappaB activity［J］. Eur J Immunol, 2009, 39(9): 2556-2570.
［14］Maeda S, Hikiba Y, Sakamoto K, Nakagawa H, Hirata Y, Hayakawa Y, et al. Ikappa B kinasebeta/nuclear factor-kappaB activation controls the development of liver metastasis by way of interleukin-6 expression［J］. Hepatology, 2009, 50(6): 1851-1860.
［15］Blackwell TS, Christman JW. The role of nuclear factor-kappa B in cytokine gene regulation［J］. Am J Respir Cell Mol Biol, 1997, 17(1): 3-9.
［16］Sethi G, Ahn KS, Aggarwal BB. Targeting nuclear factor-kappa B activation pathway by thymoquinone: role in suppression of antiapoptotic gene products and enhancement of apoptosis［J］. Mol Cancer Res, 2008, 6(6): 1059-1070.
［17］Ciucci A, Gianferretti P, Piva R, Guyot T, Snape TJ, Roberts SM, et al. Induction of apoptosis in estrogen receptor-negative breast cancer cells by natural and synthetic cyclopentenones: role of the IkappaB kinase/nuclear factor-kappaB pathway［J］. Mol Pharmacol, 2006, 70(5): 1812-1821.
［18］王永堂,鲁秀敏,李关荣. 内毒素肝损伤过程中NF-κB和AP-1活性变化及其对IL-6表达的调控［J］. 世界华人消化杂志, 2002, 10(7): 787-791.
［19］Lin ST, Wang Y, Xue Y, Feng DC, Xu Y, Xu LY. Tetrandrine suppresses LPS-induced astrocyte activation via modulating IKKs-IkappaBalpha-NF-kappaB signaling pathway［J］. Mol Cell Biochem, 2008, 315(1-2): 41-94.
［20］Jeschke MG, Barrow RE, Suzuki F, Rai J, Benjamin D, Herndon DN. IGF-I/IGFBP-3 equilibrates ratios of pro-to anti-inflammatory cytokines, which are predictors for organ function in severely burned pediatric patients［J］. Mol Med, 2002, 8(5): 238-246.