中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2013年
10期
759-763
,共5页
黄绍芳%吴飞%刘卫%贺永文
黃紹芳%吳飛%劉衛%賀永文
황소방%오비%류위%하영문
肝功能衰竭,急性%高迁移族蛋白B1%血栓调节蛋白N端结构域%可溶性晚期糖化终末产物受体
肝功能衰竭,急性%高遷移族蛋白B1%血栓調節蛋白N耑結構域%可溶性晚期糖化終末產物受體
간공능쇠갈,급성%고천이족단백B1%혈전조절단백N단결구역%가용성만기당화종말산물수체
Liver failure,acute%High mobility group box 1%The N-terminal lectin-like domain of thrombomodulin%Soluble Receptor for advanced glycation end products
目的 探索重组血栓调节蛋白N端结构域(rTM-N)和可溶性晚期糖化终末产物受体(rsRAGE)对急性肝衰竭模型小鼠的保护作用. 方法 原核表达、纯化得到rTM-N和rsRAGE,腹腔注射rTM-N或rsRAGE至急性肝衰竭模型小鼠,用不同方法检测肝组织高迁移族蛋白(HMG) B1及核因子-κ B的表达水平、血清中肿瘤坏死因子α和白细胞介素1 β的含量.对数据进行单因素方差分析. 结果 成功表达并纯化rTM-N和rsRAGE,相对分子质量约为1600和31600.各实验组小鼠肝组织坏死情况明显好转,免疫组织化学检测显示,HMGB1表达水平在实验1组和实验2组明显低于对照组；免疫荧光检测核因子-κB的表达水平,在实验1组和实验2组也明显低于对照组.RT-PCR检测HMGB1 mRNA显示,实验1组峰值(114.99±6.53)和实验2组峰值(98.44±13.69)明显低于对照组峰值(199.30±11.51),差异有统计学意义(P＜0.05).血清肿瘤坏死因子α在实验1组12h的峰值为(355.42±53.55)pg/ml、实验2组12h的峰值为(451.15±79.66)pg/ml,也显著低于对照组6h的峰值(634.23±35.18)pg/ml,差异有统计学意义(F=109.31,P＜0.05).血清白细胞介素1 β在实验l组的峰值(457.32±34.99) pg/ml和实验2组的峰值(530.66±38.83)pg/ml显著低于对照组在6h达到的峰值(688.91±48.27) pg/ml,差异有统计学意义(F=65.48,P＜0.05).结论 rsRAGE和rTM-N均可显著减少急性肝衰竭模型小鼠肝组织HMGB1的表达水平,显示对急性肝衰竭鼠较强的保护作用.
目的 探索重組血栓調節蛋白N耑結構域(rTM-N)和可溶性晚期糖化終末產物受體(rsRAGE)對急性肝衰竭模型小鼠的保護作用. 方法 原覈錶達、純化得到rTM-N和rsRAGE,腹腔註射rTM-N或rsRAGE至急性肝衰竭模型小鼠,用不同方法檢測肝組織高遷移族蛋白(HMG) B1及覈因子-κ B的錶達水平、血清中腫瘤壞死因子α和白細胞介素1 β的含量.對數據進行單因素方差分析. 結果 成功錶達併純化rTM-N和rsRAGE,相對分子質量約為1600和31600.各實驗組小鼠肝組織壞死情況明顯好轉,免疫組織化學檢測顯示,HMGB1錶達水平在實驗1組和實驗2組明顯低于對照組；免疫熒光檢測覈因子-κB的錶達水平,在實驗1組和實驗2組也明顯低于對照組.RT-PCR檢測HMGB1 mRNA顯示,實驗1組峰值(114.99±6.53)和實驗2組峰值(98.44±13.69)明顯低于對照組峰值(199.30±11.51),差異有統計學意義(P＜0.05).血清腫瘤壞死因子α在實驗1組12h的峰值為(355.42±53.55)pg/ml、實驗2組12h的峰值為(451.15±79.66)pg/ml,也顯著低于對照組6h的峰值(634.23±35.18)pg/ml,差異有統計學意義(F=109.31,P＜0.05).血清白細胞介素1 β在實驗l組的峰值(457.32±34.99) pg/ml和實驗2組的峰值(530.66±38.83)pg/ml顯著低于對照組在6h達到的峰值(688.91±48.27) pg/ml,差異有統計學意義(F=65.48,P＜0.05).結論 rsRAGE和rTM-N均可顯著減少急性肝衰竭模型小鼠肝組織HMGB1的錶達水平,顯示對急性肝衰竭鼠較彊的保護作用.
목적 탐색중조혈전조절단백N단결구역(rTM-N)화가용성만기당화종말산물수체(rsRAGE)대급성간쇠갈모형소서적보호작용. 방법 원핵표체、순화득도rTM-N화rsRAGE,복강주사rTM-N혹rsRAGE지급성간쇠갈모형소서,용불동방법검측간조직고천이족단백(HMG) B1급핵인자-κ B적표체수평、혈청중종류배사인자α화백세포개소1 β적함량.대수거진행단인소방차분석. 결과 성공표체병순화rTM-N화rsRAGE,상대분자질량약위1600화31600.각실험조소서간조직배사정황명현호전,면역조직화학검측현시,HMGB1표체수평재실험1조화실험2조명현저우대조조；면역형광검측핵인자-κB적표체수평,재실험1조화실험2조야명현저우대조조.RT-PCR검측HMGB1 mRNA현시,실험1조봉치(114.99±6.53)화실험2조봉치(98.44±13.69)명현저우대조조봉치(199.30±11.51),차이유통계학의의(P＜0.05).혈청종류배사인자α재실험1조12h적봉치위(355.42±53.55)pg/ml、실험2조12h적봉치위(451.15±79.66)pg/ml,야현저저우대조조6h적봉치(634.23±35.18)pg/ml,차이유통계학의의(F=109.31,P＜0.05).혈청백세포개소1 β재실험l조적봉치(457.32±34.99) pg/ml화실험2조적봉치(530.66±38.83)pg/ml현저저우대조조재6h체도적봉치(688.91±48.27) pg/ml,차이유통계학의의(F=65.48,P＜0.05).결론 rsRAGE화rTM-N균가현저감소급성간쇠갈모형소서간조직HMGB1적표체수평,현시대급성간쇠갈서교강적보호작용.
Objective To evaluate the roles of N-terminal lectin-like domain of thrombomodulin (TM-N) and receptor for advanced glycation end products (RAGE) in acute hepatic failure using a mouse model system.Methods Acute hepatic failure was induced in Kunming mice by intraperitonenl injection of D-galactosamine (D-Galn at 600 mg/kg) and lipopolysaccharide (LPS at 5 μg/kg) and mice were divided into groups for injection with saline,recombinant (r)TM-N protein,or recombinant soluble (rs)RAGE protein.Unmanipulated model mice served as the negative controls.Effects on liver expression of high mobility group box-1 (HMGB1) were detected by immunohistochemistry and real time RT-PCR.Effects on serum levels of tumor necrosis factor-alpha (TNFα) and interleukin-1 beta (IL)-1b were quantified by ELISA.Results Treatment with rTM-N and rsRAGE both alleviated the acute liver damage induced by D-Galn/LPS exposure,and decreased the hepatic expression of HMGB1 as well as the serum levels of TNFα and IL-1b.Conclusion Intraperitoneal delivery of rTM-N and rsRAGE can alleviate acute liver damage by modulating the expression of necrosis-and inflammatión-related factors.
