CAJ | 학술논문

目的通过动物实验研究重组纤维连接蛋白N端肝素结合域多肽(rhFNHN-29多肽)对内毒素血症肝衰竭的治疗作用.方法用内毒素脂多糖(LPS,Sigma)和半乳糖胺(Galactosamine GalN,Sigma)注射小鼠腹腔,建立内毒素血症肝衰竭小鼠模型.实验小鼠随机分两组,1组为rhFNHN-29多肽治疗组,1组为生理盐水对照组,另设正常对照组.治疗组分别于注射(LPS和GalN)前半小时,后1、2、3 h尾静脉注射rhFNHN-29多肽10 mg/kg,生理盐水对照组注射等体积生理盐水,正常对照组注射等体积生理盐水.腹腔注射药物6 h,眶静脉采血检测小鼠血浆TNFα,72 h后计算小鼠死亡率,处死活鼠,取其肝、肾、脾、肺、心、脑组织进行病理组织学观察,肝组织做电镜超微结构观察,取肝组织对其TNFα、IL-1β、IL-6的表达进行PCR分析.结果生理盐水对照组小鼠72 h死亡率为70%,多肽治疗组死亡率仅为15%.病理组织学观察显示生理盐水对照组肝组织呈广泛变性与严重坏死,多肽治疗组肝组织呈小面积变性和轻微坏死.电镜超微结构显示生理盐水埘照组肝细胞呈严重的变性和坏死,多肽治疗组轻微变性.肝细胞因子表达分析显示多肽治疗组小鼠的TNFα、IL-1β、IL-6 mRNA表达水平及血浆TNFα水平(0.86±0.43,0.86±0.31,0.27±0.13和23.6±5.8,P<0.01)显著低于生理盐水对照组小鼠的水平(1.26±0.37,0.98±0.21,0.43±0.17和87.43±16.7,P<0.01).结论 rhFNHN-29多肽对内毒素血症肝衰竭具有明显的预防和治疗作用,其作用机制可能与多肽的抗炎等作用有关.
목적 통과동물실험연구중조섬유련접단백N단간소결합역다태(rhFNHN-29다태)대내독소혈증간쇠갈적치료작용.방법 용내독소지다당(LPS,Sigma)화반유당알(Galactosamine GalN,Sigma)주사소서복강,건립내독소혈증간쇠갈소서모형.실험소서수궤분량조,1조위rhFNHN-29다태치료조,1조위생리염수대조조,령설정상대조조.치료조분별우주사(LPS화GalN)전반소시,후1、2、3 h미정맥주사rhFNHN-29다태10 mg/kg,생리염수대조조주사등체적생리염수,정상대조조주사등체적생리염수.복강주사약물6 h,광정맥채혈검측소서혈장TNFα,72 h후계산소서사망솔,처사활서,취기간、신、비、폐、심、뇌조직진행병리조직학관찰,간조직주전경초미결구관찰,취간조직대기TNFα、IL-1β、IL-6적표체진행PCR분석.결과 생리염수대조조소서72 h사망솔위70%,다태치료조사망솔부위15%.병리조직학관찰현시생리염수대조조간조직정엄범변성여엄중배사,다태치료조간조직정소면적변성화경미배사.전경초미결구현시생리염수시조조간세포정엄중적변성화배사,다태치료조경미변성.간세포인자표체분석현시다태치료조소서적TNFα、IL-1β、IL-6 mRNA표체수평급혈장TNFα수평(0.86±0.43,0.86±0.31,0.27±0.13화23.6±5.8,P<0.01)현저저우생리염수대조조소서적수평(1.26±0.37,0.98±0.21,0.43±0.17화87.43±16.7,P<0.01).결론 rhFNHN-29다태대내독소혈증간쇠갈구유명현적예방화치료작용,기작용궤제가능여다태적항염등작용유관.
Objective To study the preventive effect of recombinant polypeptide of N-terminal heparin-binding domain of fibronectin on hepatic failure induced by endotoxin in mice. Methods The 40 hepatic failure Balb/C mice were established by intraperitoneal injection of lipopolysaccharide ( LPS) and d-galactosamine(GalN). The mice were randomly divided into two groups,one for polypeptide treatment, the othe for saline treatment. Another 20 mice were used as normal control. Half hour prior to , 1,2 ,and 3 hours after injection of LPS and GalN, the rhFNHN-29 polypeptide (10 mg/kg) was injected through the tail vein of mice. The same volume of saline was given to the saline treated group and the normal control group. Six hours after the injection of LPS and GalN, 250 μl blood was taken from the eye vein of each mouse for plasma TNFα testing, and 72 hours after the injection, mortality rates of the mice of different groups were observated. The liver, lung, heart, kidney, and brain tissues of the survival mice were examined for histopathology after 72 hours . The Liver tissue was also examined for electron micrograph and for mRNA expression of TNFα, IL-1 β, IL-6 by RT-PCR. Results The 72 hours mortality rates in saline-treated and polypeptide treated-mice were 70% and 15% respectively (P < 0. 01 ) . The histopathology showed that necrosis occurred less on the hepatocytes of polypeptide treated mice than on the saline treated ones. The ultrastructure of hepatocyte under the electron microscope showed that cell apparatus of saline treated mice were destroyed and cytoplasm become loose. The expression level of TNFα,IL-l β,IL-6 mRNA on hepatocytes in polypeptide treated mice was significantly lower( 1. 26 ± 0. 37,0. 98 ± 0. 21,0. 43 ± 0. 17,87. 43 ± 16. 7 respectively) than that in the saline treated ones(1.98 ±0. 56,1. 24 ± 0. 35,0. 64 ± 0. 25 and 236. 11 ± 32.7, respectively) (P<0. 01). Similarly, the plasm TNFα level(87.43 ±16.7 ) in polypeptide treated group was significantly lower than that(236. 11 ±32. 7 ) in the saline treated group(P <0. 01). Conclusion The rhFNHN-29 polypeptide can prevent and treat hepatic failure induced by endotoxin . The mechanism by which the polypeptide takes the effect may involve its ability to down-regulate expression of those inflammative factors such as TNFα, IL-1β, IL-6.