内毒素休克大鼠脑损伤的分子机制及人参二醇组皂苷对其影响的实验研究
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摘要
通过复制大鼠内毒素休克模型,探讨内毒素引起脑损伤的分子机制,同时对比观察了人参二醇组皂苷(PDS)与地塞米松(Dex)对其的影响。主要研究发现,内毒素休克时,大鼠脑皮质CD14、TLR4/TLR2表达增加;采用电泳迁移率改变分析(EMSA)检测脑皮质转录因子NF-κB的DNA结合活性,结果显示休克后1h和4h NF-κB的DNA结合活性增高,且以1h增高明显。脑组织中特有的谷氨酸转运体表达减少(EAAT2和EAAT3),也参与了内毒素休克脑损伤。所以,可以认为脑组织细胞与其它细胞一样,NF-κB在炎症反应的早期被激活,随即诱导IL-18等细胞因子、NOS等的产生,引起脑组织损伤。谷氨酸转运体表达下降和自由基的生成、NO的大量释放存在着联系,亦可能与给予LPS后激活了NF-κB有关。
本研究发现PDS 具有改善内毒素休克大鼠血液动力学、流变学和抗氧化损伤作用;还可降低脑细胞CD14、TLR4/TLR2 的表达。我们采用EMSA实验首次证实PDS 可明显抑制NF-κB 的DNA 结合活性,从而减少细胞因子的产生。与Dex 相比,PDS 通过影响脑细胞内毒素信号转导途径的多个环节,具有多靶点细胞保护作用。
21st century is a “Brain’s Era”. For a long time , it is only a wonderful dream for human being to explain the brain clearly because of the complexity of the cerebral structures and functions. In 1996,the Japanese scientists presented the slogan that “understanding brain”“protecting brain”“developing brain”. The Noble Prize winner Watson Crick said:“There is no more important research than the study on the brain for our human being.”IBRO(International Brain Research Organization) proposed that the 21st century is a “Brain’s Era”. In general, brain research has become one of the most popular topics in this century.
Lipopolysaccharide(LPS) is the main component of endotoxin(ET) which is released from Gram negative bacteria after death or during the rapid growing course. As a pathogen, LPS exists extensively in the nature. After LPS entering the body, many pathophysiological reactions can take place, such as fever, dilation of blood vessels, vascular permeation increase, leukocytosis, complement activation,
blood pressure decrease and etc. In some serious cases, LPS can also lead to disseminated intravascular coagulation(DIC), fatal shock and multiple organ dysfunction syndrome(MODS). Above all, LPS plays a very important role on the pathogenesis of Gram negative bacterial infection. With the development of basic medicine and clinical research, people have some insight of the structure, function, and action mechanism of LPS. But now the most study of the diseases caused by LPS is focused and limited on peripheral organs such as lung, heart, liver, kidney, and et.al. The corresponding knowledge about brain is little known by now. There are many suspending problems and many different views in the nervous system diseases because of the existence of brain barrier and the complicated structures. Scientists always think that endotoxin is too large to pass through the brain barrier, but some scientists believe that big dose of LPS injection from peripheral blood can come into the brain by passing through the brain barrier.
At present , we still can not deny the possibility when the brain barrier is destroyed by diseases or big dose of LPS can enter hypothalamus and act on the neuronal cells. The recent research demonstrates that peripheral LPS injection can cause a series of intracerebral changes such as iNOS activity increase in the astrocytes which involved in the pathological process of CNS bacterial infection. At the same time, LPS can also stimulate the astrocytes to secrete IL-1、TNF-αand IL-6 which enhanced the inflammation injury of the brain. Furthermore there are still many
different even paradoxical results. Based on such backgrounds, this experiment has explored the oxidization injury of endotoxin shock rats cerebral cortex, endotoxin signal transduction pathway and the changes of NF-κB and glutamate transporter expression. Through the study of the LPS action mechanism on central nervous system, we aimed to look for the effective target medicine for treating endotoxin disease . On such background, we duplicated the animal models of endotoxic shock rats to reveal the molecular mechanism of cerebral injury by LPS and observe the Panaxadiol(PDS) effects as well.
The results of this experimental research are as follows:
1. Endotoxic shock model: One hundred Wistar rats were divided into 5 groups randomly the experimental control group (Control group); the LPS endotoxic shock model group (LPS group); Dexamethasone administered group (LPS+DEX group); PDS low-dose group (LPS+PDSL group, 22.5mg/Kg); PDS mid-dose group (LPS+PDSM group,45.0mg/kg); PDS high-dose group (LPS+PDSH group,90.0mg/kg), and each group was divided into 2 groups: killed at 60min and killed at 240min after shock. LPS (4mg/kg) was sublingually administered intravenously to build up the animal models of endotoxic shock. In addition, among the PDS-treated groups, PDS was administered before the use of LPS. When blood pressure fell to 2/3 of the baseline it was regarded as the start point of shock. The mean arterial blood pressure (MABP) was monitored dynamically by the RM6000 4-lead physiologic recording apparatus. Meanwhile, the blood viscosity was determined, and
histomorphological changes of brain tissue was observed as well.
(1) MABP dynamic observation: Injection of LPS, rats MABP fell to 2/3 of the baseline in all groups after 5 minutes and 10 minutes later, MABP fell progressively. MABP had an compensatory restoration at 20min of shock and at 30min reached the summit. From then on, the above index come down constantly again until the models got close to death at 240min.
Though the other groups treated with PDS took on the similar effects after injection of LPS , 30-60 minutes later they had steady and constant effect of increasing MABP which is not like that of the groups treated with only LPS. The statistical analysis demonstrated that MABP in the groups treated with PDS was significantly higher than that of LPS groups at the time of 60min, 120min, 180min and 240min after shock (p<0.05,p<0.01,p<0.001). At the same time, there was no significance of MABP between PDS groups and Dex groups(p>0.05).
The LPS endotoxic shock model duplicated successfully on the evidence that the decreasing percentage of MABP at the different time is accordance to what of general MABP. This gave off the individual variance interference and set up a good basis for further study.
(2) Observation on pathological alternation of cerebral tissue.
In LPS group brain tissue we observed that inflammatory cells infiltration, peri-capillary space increase, partial neuronal
vacuolar degeneration, karyopyknosis, obvious contraction of Nissl's body and decrease of neurone amount. The tendency is dependant on stimulation time. Otherwise in LPS+Dex group, LPS+PDSL group, LPS+PDSM group and LPS+PDSH group, the degree of this kind of injury alternation was less relatively.
(3) Microcirculation change is one of most important indicators reflecting shock state. Some people even thought shock is almost a hema-rheological phenomenon. : the blood viscosity in PDS groups are all significantly higher under various shear rates than that in other groups(p<0.05). It suggested that shock had been in out of compensatory and irreversible stage.
(4) Result of the determination of LPO and NO contents, activity of SOD and NOS:
The activity of SODs in Control group, LPS+Dex group, LPS+PDSLgroup and LPS+PDSM is significantly higher than that in LPS group(p<0.05) after 4 hours of shock. However LPO contents in former groups are significantly lower than that in LPS group. At the same time, the activity of NOSs and NO contents in Control group, LPS+Dex group, LPS+PDSL group and LPS+PDSM are significantly lower than those in LPS group(p<0.05) after 4 hours of shock.
2.The molecular mechanism of brain injury caused by LPS-induced endotoxic shock and the protective effects of PDS
The precondition to express LPS biological effect is LPS binding corresponding receptors on the membrane. CD14 and Toll-like receptors(TLRs) are two very important model-recognizing receptors which play a important role on resistance of various pathogen.
(1) The expression of CD14 mRNA and protein on cerebral cortex: In normal situation, the expression of CD14 mRNA and protein on cerebral cortex is not very strong. By the time of the LPS stimulation, results of CD14 mRNA expression and the expression of CD14 protein with western blot showed that in LPS group is significantly higher than that in control group after 4 hours of shock( p<0.05). While in DEX group and PDS groups, the expression of CD14 mRNA decreased significantly(p<0.05 vs LPS group).
(2) The TLRs family mRNA expression changes on cerebral cortex: The experiment showed that, with the LPS stimulation time, TLR4 and TLR2mRNA expression increased constantly and significantly. While in LPS group, TLR9mRNA expression increased but had no significance versus Control group( p<0.05). However PDS could inhibit TLR4/TLR2mRNA expression.
(3) IL-18 changes on cerebral cortex of LPS-induced endotoxic shock rats: The experimental results demonstrated that IL-18mRNA with RT-PCR and IL-18 protein expression with Western blot in LPS groups were significantly higher than those in Control group at the time of 1h and 4h after shock(p>0.05). But Dex and PDS could lessen the expression of them at some degree.
3.The expression of NF-κB/IκB on cerebral cortex
In this study, we utilized EMSA method to detect NF-κB binding activity with DNA, use Western Blot to determine the expression of NF-kB subunit protein and use immunochemical method to observe the nuclear translocation process of NF-κB. Most current data shows
that NF-κB will be activated by stimulation after 30min, and reach peak after 1h.Thereafter, we kill the rats at the time of 1h and 4h to observe the NF-κB change.
EMSA result showed that NF-κB binding activity with DNA significantly strengthened after LPS injection. In this experiment we extracted nuclear protein and cytosolic protein to detect the NF-κB subunit protein expression. Contrast to Control group, p65 of nuclear protein expression with Western blot was strengthened after 1h and 4h LPS injection(p<0.05)and the same result happened to p50 protein. In LPS+Dex group, LPS+PDSL group and LPS+PDSM group, p65 protein expression decreased significantly but p50 protein expression decreased not obviously. These results was accordance to that of EMSA. The research also showed that before and after LPS injection , the cytosol p50 and p65 protein expressed and not decreased significantly because of the p65/p50 nuclear translocation among different groups (p>0.05).
The immunochemistry results demonstrated that the protein of p65 and p50 for Control group expressed mainly in cytoplasm of neuron. However they translocated into nucleus after LPS stimulation, and the immunochemistry showed the process clearly. In this study, we proved that the NF-kB was mediated by binding with I-kB and in LPS group I-kB mRNA expression was less than Control group obviously(p<0.05).
4.EAAT expression on cerebral cortex:
In contrast to Control group after LPS injection, EAAT2mRNA expression decreased at 1h and 4h(p<0.05); But EAAT3mRNA only
decreased a little without significance at 1h (p>0.05) whereas decreased obviously at 4h (p<0.05). The further Western blot results showed the same changing tendency of EAAT2 and EAAT3 protein expression. There were no obvious changes of EAAT1和EAAT4mRNA expression after LPS stimulation (p>0.05). in addition, EAAT3 protein expression increased in LPS+Dex group, LPS+PDSL group and LPS+PDSM group at 4h. The immunochemistry results demonstrated EAAT2 located on astrocytic membrane while EAAT3 expressed on neuronal membrane.
In conclusion, during endotoxic shock, LPS provoked LPS/CD14 complex expression, and then activated TLR4/TLR2,in turn affected a series of subsequent protein. After phosphorylation of IkB which induced the translocation of NF-kB to the nucleus, NF-kB associated with specific DNA binding sites initiating gene transcription. It induced the expression of mRNA of a variety of pro-inflammatory mediators including iNOS/NO 、and IL-18. This process finally becomes the major cause to the brain tissue injury. The brain as an integrating nervous regulating organ, its functional alternation and structural injury will affect other organ’s response to cytokines which canconfer that LPS may regulate the LPS/CD14/TLR4/NF-κB expression and some inflammatory cytokines release to cause cerebral injury and shock condition. In addition, other study shows that oxygen free radicals also can be some kinds of signal molecules to mediate NF-kB signaling pathway which involves the cell damage. Our results proved the brain EAATmRNA expression decrease after LPS injection associated with the free
本研究发现PDS 具有改善内毒素休克大鼠血液动力学、流变学和抗氧化损伤作用;还可降低脑细胞CD14、TLR4/TLR2 的表达。我们采用EMSA实验首次证实PDS 可明显抑制NF-κB 的DNA 结合活性,从而减少细胞因子的产生。与Dex 相比,PDS 通过影响脑细胞内毒素信号转导途径的多个环节,具有多靶点细胞保护作用。
21st century is a “Brain’s Era”. For a long time , it is only a wonderful dream for human being to explain the brain clearly because of the complexity of the cerebral structures and functions. In 1996,the Japanese scientists presented the slogan that “understanding brain”“protecting brain”“developing brain”. The Noble Prize winner Watson Crick said:“There is no more important research than the study on the brain for our human being.”IBRO(International Brain Research Organization) proposed that the 21st century is a “Brain’s Era”. In general, brain research has become one of the most popular topics in this century.
Lipopolysaccharide(LPS) is the main component of endotoxin(ET) which is released from Gram negative bacteria after death or during the rapid growing course. As a pathogen, LPS exists extensively in the nature. After LPS entering the body, many pathophysiological reactions can take place, such as fever, dilation of blood vessels, vascular permeation increase, leukocytosis, complement activation,
blood pressure decrease and etc. In some serious cases, LPS can also lead to disseminated intravascular coagulation(DIC), fatal shock and multiple organ dysfunction syndrome(MODS). Above all, LPS plays a very important role on the pathogenesis of Gram negative bacterial infection. With the development of basic medicine and clinical research, people have some insight of the structure, function, and action mechanism of LPS. But now the most study of the diseases caused by LPS is focused and limited on peripheral organs such as lung, heart, liver, kidney, and et.al. The corresponding knowledge about brain is little known by now. There are many suspending problems and many different views in the nervous system diseases because of the existence of brain barrier and the complicated structures. Scientists always think that endotoxin is too large to pass through the brain barrier, but some scientists believe that big dose of LPS injection from peripheral blood can come into the brain by passing through the brain barrier.
At present , we still can not deny the possibility when the brain barrier is destroyed by diseases or big dose of LPS can enter hypothalamus and act on the neuronal cells. The recent research demonstrates that peripheral LPS injection can cause a series of intracerebral changes such as iNOS activity increase in the astrocytes which involved in the pathological process of CNS bacterial infection. At the same time, LPS can also stimulate the astrocytes to secrete IL-1、TNF-αand IL-6 which enhanced the inflammation injury of the brain. Furthermore there are still many
different even paradoxical results. Based on such backgrounds, this experiment has explored the oxidization injury of endotoxin shock rats cerebral cortex, endotoxin signal transduction pathway and the changes of NF-κB and glutamate transporter expression. Through the study of the LPS action mechanism on central nervous system, we aimed to look for the effective target medicine for treating endotoxin disease . On such background, we duplicated the animal models of endotoxic shock rats to reveal the molecular mechanism of cerebral injury by LPS and observe the Panaxadiol(PDS) effects as well.
The results of this experimental research are as follows:
1. Endotoxic shock model: One hundred Wistar rats were divided into 5 groups randomly the experimental control group (Control group); the LPS endotoxic shock model group (LPS group); Dexamethasone administered group (LPS+DEX group); PDS low-dose group (LPS+PDSL group, 22.5mg/Kg); PDS mid-dose group (LPS+PDSM group,45.0mg/kg); PDS high-dose group (LPS+PDSH group,90.0mg/kg), and each group was divided into 2 groups: killed at 60min and killed at 240min after shock. LPS (4mg/kg) was sublingually administered intravenously to build up the animal models of endotoxic shock. In addition, among the PDS-treated groups, PDS was administered before the use of LPS. When blood pressure fell to 2/3 of the baseline it was regarded as the start point of shock. The mean arterial blood pressure (MABP) was monitored dynamically by the RM6000 4-lead physiologic recording apparatus. Meanwhile, the blood viscosity was determined, and
histomorphological changes of brain tissue was observed as well.
(1) MABP dynamic observation: Injection of LPS, rats MABP fell to 2/3 of the baseline in all groups after 5 minutes and 10 minutes later, MABP fell progressively. MABP had an compensatory restoration at 20min of shock and at 30min reached the summit. From then on, the above index come down constantly again until the models got close to death at 240min.
Though the other groups treated with PDS took on the similar effects after injection of LPS , 30-60 minutes later they had steady and constant effect of increasing MABP which is not like that of the groups treated with only LPS. The statistical analysis demonstrated that MABP in the groups treated with PDS was significantly higher than that of LPS groups at the time of 60min, 120min, 180min and 240min after shock (p<0.05,p<0.01,p<0.001). At the same time, there was no significance of MABP between PDS groups and Dex groups(p>0.05).
The LPS endotoxic shock model duplicated successfully on the evidence that the decreasing percentage of MABP at the different time is accordance to what of general MABP. This gave off the individual variance interference and set up a good basis for further study.
(2) Observation on pathological alternation of cerebral tissue.
In LPS group brain tissue we observed that inflammatory cells infiltration, peri-capillary space increase, partial neuronal
vacuolar degeneration, karyopyknosis, obvious contraction of Nissl's body and decrease of neurone amount. The tendency is dependant on stimulation time. Otherwise in LPS+Dex group, LPS+PDSL group, LPS+PDSM group and LPS+PDSH group, the degree of this kind of injury alternation was less relatively.
(3) Microcirculation change is one of most important indicators reflecting shock state. Some people even thought shock is almost a hema-rheological phenomenon. : the blood viscosity in PDS groups are all significantly higher under various shear rates than that in other groups(p<0.05). It suggested that shock had been in out of compensatory and irreversible stage.
(4) Result of the determination of LPO and NO contents, activity of SOD and NOS:
The activity of SODs in Control group, LPS+Dex group, LPS+PDSLgroup and LPS+PDSM is significantly higher than that in LPS group(p<0.05) after 4 hours of shock. However LPO contents in former groups are significantly lower than that in LPS group. At the same time, the activity of NOSs and NO contents in Control group, LPS+Dex group, LPS+PDSL group and LPS+PDSM are significantly lower than those in LPS group(p<0.05) after 4 hours of shock.
2.The molecular mechanism of brain injury caused by LPS-induced endotoxic shock and the protective effects of PDS
The precondition to express LPS biological effect is LPS binding corresponding receptors on the membrane. CD14 and Toll-like receptors(TLRs) are two very important model-recognizing receptors which play a important role on resistance of various pathogen.
(1) The expression of CD14 mRNA and protein on cerebral cortex: In normal situation, the expression of CD14 mRNA and protein on cerebral cortex is not very strong. By the time of the LPS stimulation, results of CD14 mRNA expression and the expression of CD14 protein with western blot showed that in LPS group is significantly higher than that in control group after 4 hours of shock( p<0.05). While in DEX group and PDS groups, the expression of CD14 mRNA decreased significantly(p<0.05 vs LPS group).
(2) The TLRs family mRNA expression changes on cerebral cortex: The experiment showed that, with the LPS stimulation time, TLR4 and TLR2mRNA expression increased constantly and significantly. While in LPS group, TLR9mRNA expression increased but had no significance versus Control group( p<0.05). However PDS could inhibit TLR4/TLR2mRNA expression.
(3) IL-18 changes on cerebral cortex of LPS-induced endotoxic shock rats: The experimental results demonstrated that IL-18mRNA with RT-PCR and IL-18 protein expression with Western blot in LPS groups were significantly higher than those in Control group at the time of 1h and 4h after shock(p>0.05). But Dex and PDS could lessen the expression of them at some degree.
3.The expression of NF-κB/IκB on cerebral cortex
In this study, we utilized EMSA method to detect NF-κB binding activity with DNA, use Western Blot to determine the expression of NF-kB subunit protein and use immunochemical method to observe the nuclear translocation process of NF-κB. Most current data shows
that NF-κB will be activated by stimulation after 30min, and reach peak after 1h.Thereafter, we kill the rats at the time of 1h and 4h to observe the NF-κB change.
EMSA result showed that NF-κB binding activity with DNA significantly strengthened after LPS injection. In this experiment we extracted nuclear protein and cytosolic protein to detect the NF-κB subunit protein expression. Contrast to Control group, p65 of nuclear protein expression with Western blot was strengthened after 1h and 4h LPS injection(p<0.05)and the same result happened to p50 protein. In LPS+Dex group, LPS+PDSL group and LPS+PDSM group, p65 protein expression decreased significantly but p50 protein expression decreased not obviously. These results was accordance to that of EMSA. The research also showed that before and after LPS injection , the cytosol p50 and p65 protein expressed and not decreased significantly because of the p65/p50 nuclear translocation among different groups (p>0.05).
The immunochemistry results demonstrated that the protein of p65 and p50 for Control group expressed mainly in cytoplasm of neuron. However they translocated into nucleus after LPS stimulation, and the immunochemistry showed the process clearly. In this study, we proved that the NF-kB was mediated by binding with I-kB and in LPS group I-kB mRNA expression was less than Control group obviously(p<0.05).
4.EAAT expression on cerebral cortex:
In contrast to Control group after LPS injection, EAAT2mRNA expression decreased at 1h and 4h(p<0.05); But EAAT3mRNA only
decreased a little without significance at 1h (p>0.05) whereas decreased obviously at 4h (p<0.05). The further Western blot results showed the same changing tendency of EAAT2 and EAAT3 protein expression. There were no obvious changes of EAAT1和EAAT4mRNA expression after LPS stimulation (p>0.05). in addition, EAAT3 protein expression increased in LPS+Dex group, LPS+PDSL group and LPS+PDSM group at 4h. The immunochemistry results demonstrated EAAT2 located on astrocytic membrane while EAAT3 expressed on neuronal membrane.
In conclusion, during endotoxic shock, LPS provoked LPS/CD14 complex expression, and then activated TLR4/TLR2,in turn affected a series of subsequent protein. After phosphorylation of IkB which induced the translocation of NF-kB to the nucleus, NF-kB associated with specific DNA binding sites initiating gene transcription. It induced the expression of mRNA of a variety of pro-inflammatory mediators including iNOS/NO 、and IL-18. This process finally becomes the major cause to the brain tissue injury. The brain as an integrating nervous regulating organ, its functional alternation and structural injury will affect other organ’s response to cytokines which canconfer that LPS may regulate the LPS/CD14/TLR4/NF-κB expression and some inflammatory cytokines release to cause cerebral injury and shock condition. In addition, other study shows that oxygen free radicals also can be some kinds of signal molecules to mediate NF-kB signaling pathway which involves the cell damage. Our results proved the brain EAATmRNA expression decrease after LPS injection associated with the free
引文
[1]张顺财主编. 内毒素基础与临床[M]. 北京,科学出版社,2003:1-436.
[2]罗海波,鲍行豪. 细菌内毒素研究进展(第一版)[M]. 北京,人民卫生出版社,1983:1-373.
[3]Bayston KF, Cohen J. Bacterial endotoxin and current concepts in the diagnosis and treatment of endotoxamia[J]. J Med Microbiol,1990,31(4):73-83.
[4]Mitov IG, Terziiski DG. Immunoprophyiaxis and immunotherapy of gram-negative sepsis and shock with antibodies to core glycolipids and lipid A of bacterial lipopolysaccha rides[J]. Infection,1991,19(10):383-390.
[5] 邓瑞麟译. 医学微生物学( 第一版)[M]. 北京, 人民卫生出版社,1983:270-272.
[6]邱世翠,曲云,胡涛英,等. 脂多糖致肺血管内巨噬细胞形态和功能的改变[J]. 滨州医学院学报,1992,15(3):211-212.
[7]Birgitt Stein, Petra Frank, Wilhelm S, et al. Endotoxin and cytokines induce direct cardiodepressive effects in mammalian cardiomyocytes via induction of nitric oxide synthase[J]. J Mol Cell Cardiol,1996,28(7):1631–1639.
[8]Torre-Amione G, Kapadia S, Lee J, et al. Tumor necrosis factor-alpha and tumor necrosis factor receptors in the failing human heart[J]. Circulation,1996,93(12):704-711.
[9]Roig E, Orus J, Pare C,et al. Serum interleukin-6 in congestive heart failure secondary to idiopathic dilated cardiomyopathy [J]. Am J Cardiol,1998,82(10):688-690.
[10]Hotchkiss, RS, Swanson PE, Freeman BD, et al. Apoptotic cell death in patients with sepsis, shock and multiple organ dysfunction[J]. Crit Care Med,1999,27(9):1230-1251.
[11]Ayala A, Xu YX, Ayala CA, et al. Increased mucosal B-lymphocyte apoptosis during polymicrobial sepsis is a Fas ligand but not an endotoxin mediated process[J]. Blood,1998, 91(10): 1362-1372.
[12]Alfred Ayala, Joanne L, Patricia S,et al. Pathological aspects of apoptosis in severe sepsis and shock[J]? The International Journal of Biochemistry & Cell Biology,2003,35(6):7-15.
[13]Francoise MA, Luc, DL, Adjuvants[J]. Immunology Today,1993, 14(6):281-293.
[14]李桂杰,朱瑞良,郭宝聚. 细菌内毒素的研究进展[J]. 山东农业大学学报,1998,30(2):193-198.
[15]陈越,金久善. 自由基的生物学效应[J]. 中国兽医杂志,1996, 22(2):48-51.
[16]Remick D. Applied molecular biology of sepsis[J]. J Crit Care,1995,10(4):198-212.
[17]Cowan DB, Poutias DN, Del PJ, et al. CD14-independent activation of cardiomyocyte signal transduction by bacterial endotoxin[J]. Am J Physiol Heart Circ Physiol, 2000, 279(2): 619-629.
[18]Martin TR. Recognition of bacterial endotoxin in the lungs[J]. Am J Respir Cell Mol Biol,2000,23(5):128-132.
[19]Tobias PS, Soldau K, Ulevitch RJ. Isolation of a lipopolysaccharide binding acute phase reactant from rabbit serum[J]. J Exp Med,1986,164(10):777-793.
[20]Heumann D, Roger T. Initial responses to endotoxins and Gram-negative bacteria[J]. Clin Chimica Acta,2002,323(8): 59-72.
[21]李永旺,麻莉,毛宝龄,等. 内毒素诱导的TLR4-MD2信号传导通路[J]. 中国药理学通报,2002,18(5):121-124.
[22]Derek S,J ong S, Edward A. Sepsis:current concepts in intracellular signaling[J]. Int J Biochem,2002,1302(3):1-7.
[23]Leanne P, Siamon C. The function of scavenger receptors expressed by macrophages and their role in the regulation of inflammation[J]. Micro infect,2001,3(1):149-159.
[24]Aderem A, Ulevitch RJ, Kawai T, et al. Toll-like receptors in the induction of the innate immune response[J]. Nature,2000, 406:782-787.
[25]Hemmi H, Takeuchi O, Kawai T, et al. A Toll-like receptor recognizes bacterial DNA[J]. Nature,2000,408:740.
[26]Medzhitov R, Preston P, Janeway CA, et al. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature,1997,388:394-397.
[27]Poltorak A, Ricciardi P, Citterio S, et al. Physical contact between lipopolysaccharide and Toll-like receptor 4 revealed by genetic complementation [J]. Proc Natl Acad Sci,2000,97: 2163.
[28]Ohashi K, Burkart V, Floh S, et al. Heat shock protein 60 is a putative endogenous ligand of the Toll-like receptor 4 complex[J]. J Immunol,2000,164:558-561.
[29]Lien E, Seiiati TJ, Yoshimura A, et al. Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products[J].Chem,1999,274(47):33419-33425.
[30]Yang RB, Mark MR, Gumey AL, et al. Signaling events induced by lipopolysaccharide-activated toll-like receptor 2[J]. J Immumol,1999,163(2):639-643.
[31]Bauer S, Kirschning CJ, Hacker H, et al. Human TLR9 corrfers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci,2001,98(16):9237-9242.
[32]Hemmi H, Takeuchi O, Kawai T, et al. A Toll-like receptor recognizes bacterial DNA[J]. Nature,2000,408:740-745.
[33]Takeuchi O, Kaufmann A,Grote K, et al. Preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-activates immune cells through a Toll-like receptor 2 and MyD88-dependent signaling pathway[J]. J Immunol,2000,164:554-557.
[34]Siebenlist U, Franzoso G, Brown K, et al. Structure, regulation and function of NF-κB[J]. Annu Rev Cell Biol,1994,10:405-455.
[35]Baeuerie PA, Baltimore D. NF-κB: ten years after[J]. Cell,1996,87(1):13-20.
[36] Karin M, Ben-Netiah Y. Phosphorylation meets ubiquitination: the control of NF-κB activity[J]. Annu Rev Immunol,2000, 18:621-663.
[37]Latimer M, Ernst MK, Dunn LL, et al. The N-Terminal domain of IκB masks the nuclear localization signals of p50 and c-Rel homodimers[J]. Mol Cell Bio,1998,18:2640-2649.
[38]Korner M, Rattner A, Mauxion F, et al. A brain-specific transcription activator. Neuron,1989,3:563-572.
[39]Mattson MP, Camandola S. NF-κB in neuronal plasticity and neurodegenerative disorders[J]. J Clin invest,2001, 107(3): 247-254.
[40]Ferret I, Marti E, Lopez E, et al. NF-κB immuno reactivity is observed in association with beta A4 diffuse plaques in patients with Alzheimer's disease.Neuropathol Appl Neurobiol, 1998, 24 (4): 271 -277.
[41]eck S, Lezoualc'h F, Engert S, et al. Insulin-like growth factor-l-mediated neuroprotection against oxidative stress is associated with activation of nuclear factor kappa B. J Biol Chem, 1999, 274(14):9828-9835.
[42]Ginn-Pease ME, Whisler RL. Redox signals and NF-κB activation in T cells . Free Radic Biol Med, 1998, 25(3):346-361.
[43]Ginn-Pease ME, Whisler RL. Redox signals and NF-κB activation in T cells . Free Radic Biol Med, 1998, 25(3):346-361.
[44]Yamamoto Y, Gaynor RB. Therapeutic potential of inhibition of the NF-κB pathway in the treatment of inflammation and cancer[J]. J Clin Invest,2001,107(2):135-142.
[45]Demling RH, Lalonde C. Relationship between lung injury and lung lipid-peroxidation caused by recurrent endotoxin[J]. Am J RevRespir D,1989,139(9):1118-1124.
[46]Sakaguchi S, Kanda N, Hsu CC, et al. Lipid peroxide for-mation and membrane damage in endotoxin-poisoned mice[J]. Microbiol Immunol,1981,25(3):229-244.
[47]Chambers DE, Parks DA, Patterson G, et al.Xanthineoxidaseas a sourse of free radical damage in myocardial ischemia[J]. J Mol Cell Cardiol,1985,17(8):145-154.
[48]Beckman JK. The role of phospholipase: A activity in rat liver microsomal lipidperoxidation[J]. J Biol Chem,1987, 262(4): 1479-1484.
[49]邱海波,陈德昌. 一氧化氮生物作用的两面性与多器官功能障碍综合征[J]. 基础医学与临床,1998,18(1):11-14.
[50]Thiemermann C, Vane JR. Inhibition of nitric oxide synthesis reduces the hypotension induced by bacterial lipopolysacch-aride in the rat[J]. Eur J Pharmacol,1993,182(11):591-595.
[51]Julou-Schaeffer G, Gray G, Fleming, et al.Loss of vascular responsiveness induced by endotoxin involves the L-arginine pathway[J]. Am. J. Physiol.,1991,259(9):H1038-H1043.
[52]Rees DD, Cellek S, Pahner RM, et al. Dexamethasone prevents the induction of nitric oxide synthase and the associated effects on the vascular tone: an insight into endotoxic shock[J]. Biochem Biophys Res Commun,1990,173(11):541-547.
[53]Kilbourn RG, Juburan A, Gross SS, et al. Reversal of endotoxin-mediated shock by N-monomethyl -L-arginine, an
inhibitor of nitric oxide synthesis[J]. Biochem Biophys Res
Commun,1990,172(2):1132-1138.
[54]Kilbourn RG, Gross SS, Jubran A, et al. N-methyl-L-arginine inhibits tumour necrosis factor-induced hypotension: implications for the involvement of nitric oxide[J]. Proc Natl Acad Sci USA,1990,87(7):3629-3632.
[55]Billiar TR, Curran RD, Harbrecht BG, et al. Modulation of nitrogen oxide synthesis in vivo: NG-monomethyl-L-arginine inhibits endotoxin-induced nitrite/nitrate biosynthesis while promoting hepatic damage[J]. J Leukoc Biol,1990,48(12): 565-569.
[56]高洪,陈利平,唐兆新. 内毒素诱生自由基致膜损伤机理的研究进展[J]. 动物医学进展,1999,20(3):1-5.
[57]Lowenstein CJ, Glatt CS, Bredt DS, et al. Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme[J]. Proc Natl Acad sci USA,1992,89(8):6711-6715.
[58]Rockey DC, Chung JJ. Regulation of inducible nitric oxide synthase in hepatic sinusoidal endothelial cells[J]. Am J physiol,1996,271(10):260-267.
[59]Curran RD, Billiar TR, Stuehr DJ, et al. Multiple cytokines are required to induce hepatocyle nitric oxide production and inhibit total protein synthesis[J]. Ann Surg,1990,212(5): 462-469.
[60]Helyar L, Bundschuh DS, Lskin JD. Induction of hepatic Ito cell nitric oxide production after acute endoxoxemia[J]. Hepatology,
1994,20(2):1509-1515.
[61]谢平初,陆家齐,孙大铭,等. 猪内毒素休克外周与内脏微循环灌注的动态变化[J]. 中国危重病急救医学,1999,11(12):718-720.
[62]Palmer RM, Ferrige A, Mocada,S et al. Nitric oxide release accounts for the biological activity of endothelium relaxing factor[J]. Nature,1987,327(12):524-530.
[63]Matute C, Alberdi E, Ibarretxe G, et al. Excitotoxicity in glial cells[J]. Eur J Pharmacol,2002,447:239-246.
[64]Schousboe A. Transport and metabolism of glutamate and GABA in neurons are glial cells[J]. Int Rev Neurobiol,1981,22:1-45.
[65]Matsushita Y, Shima, Nawashiro H, et al. Real-time monitoring of glutamate following fluid percussion brain injury wigh hypoxia in the rat[J]. J Neurotrauma,2000,17(2):143-153.
[66]Fairman WA, Vandenberg RJ, Arriza,JL, et al. Gluamate uptake[J]. Nature,1995,375:599-603.
[67]Ye Z, Sontheimer H. Modulation of glial glutamate transport through cell interactions with the extracellular matrix[J]. Int J Dev Neurosci,2002,20:209-214.
[68]Maragakis NJ, Rothstein JD. Glutamate transporters in neurologic disease. Arch Neurol,2001,58:365-370.
[69]Seal RP, Amara SG. Excitatory amino acid transperters: a family in flux[J]. Annu Rev Pharmacol Toxicol,1999,39:431-456.
[70]Niels C, Danbolt. Glutamate uptake[J]. Prog Neurobiol,2001,65: 100-105.
[71]Schluter K, Figiel M, Rozyczka J, et al. CNS region –specific regulation of glial glutamate transporter expression[J]. Eur J Neurosci,2002,16:836-842.
[72]OShea R. Roles and regulation of glutamate transporters in the central nervous system[J]. Clin Exp Pharmacol Physiol, 2002,29:1018-1023.
[73]Auger C, Attwell D. Fast removal of synaptic glutamate by postsynaptic transporters[J]. Neuron,2000,28:547-558.
[74]Bridges RJ, Kavanaugh MP, Chamberlin AR. A pharmacological review of competitive inhibitors and subatrates of high-affinity, sodium-dependent glutamate transport in the central nervous system[J]. Curr. Pharm Des,1999,5:363-379.
[75]Christopher M, Anderson, Raymond A, et al. Astrocyte glutamate transport: Review of properties, regulation, and physiological functions[J ]. Glia,2000,32:1-14.
[76]Had-Aissouni L, Re DB, Nieoullon A. Importance of astrocytic inactivation of synaptically released glutamate for cell survival in the central nervous system-are astrocytes vulnerable to low intracellular glutamate concentrations[J]? J Physiol Paris,2002,96:317-322.
[77]Kanai Y, Smith CP, Hediger MA. A new family of neurotransmitter transporters: the high-affinity glutamate transporters [J].FASEB J,1993,7(15):1450-1459.
[78]Denis F, Daignan B. Synthesis of glutamine, glycine and 10-formyl tetrahydrofolate is coregulated with purine biosynthesis in Saccharomyces cerevisiae[J].Mol Gen
Genet,1998,259(3):246-255.
[79]Nicholas J, Maragakis, Jeffrey D, et al. Glutamate transporters in neurologic disease[J]. Basic Sci Semin Neurol,2001, 58:365-370.
[80]Volterra A, Trotti D, Tromba C, et al. Glutamate uptake inhibition by oxygen free radicals in rat cortical astrocytes[J]. J Neurosci,1994,14:2924-2932.
[81]Fine SM, Angel RA, Perry SW, et al. Tumor necrosis factor alpha inhibits glutamate uptake by primary human astrocytes[J]. J Biol Chem,1996,271:15303-15306.
[82]Liao SL, Chen CJ. Differential effects of cytokines and redox potential on glutamae uptake in rat cortical glial cultures[J]. Neurosci Lett,2001,299:113-116.
[83]Wang Z, Pekarskaya O, Bencheikh M, et al. Reduced expression of glutamate transporter EAAT2 and impaired glutamate transport in human primary astrocyes exposed to HIV-1 gp 123[J]. Virology,2003,312:60-73.
[84]Pogun S, Dawson V, Kuhar MJ. Nitric oxide inhibits 3H-glutamate transport in synaptosomes[J]. Synapse,1994,18(1):21-26.
[85]Lonart G, Johnson KM. Inhibitory effects of nitric oxide on the uptake of [3H]dopamine and [3H]glutamate by striatal synaptosomes[J]. J Neurochem,1994,63(6):2108-2117.
[86]Trotti D, Aoki M, Pasinelli P. Amyotrophic lateral sclerosis-linked glutamate transporter mutant has impaired glutamate clearance capacity[J]. J Biol Chem,2001,276:576-
582.
[87]Hu SX, Sheng WS, Ehrlich LC, et al. Cytokine effects on glutamate uptake by human astrocytes[J]. Neuroimmunomod-ulation,2000,7:153-159.
[88]Piani D, Frei K, Pfister HW, et al. Glutamate uptake by astrocytes is inhibited by reactive oxygen intermediates but not by other macrophage-derived molecules including cytokines, leukotrienes or platelet-activating factor[J].J Neuroimmunol,1993,48(1):99-104.
[89]全竹富. 抗内毒素核心糖脂抗体:防治革兰氏阴性菌感染的研究现状[J]. 国外医学·外科学分册,1995,22(5):284-288.
[90]Elsbach P, Weiss J, Doerfler M, et al. The bactericidal/ permeability increasing protein of neutrophils is a potent antibacterial and anti-endotoxin agent in vitro and in vivo[J]. Prog Clin Biol Res,1994,388(2):41-54.
[91]Natanson C, Hoffmea WD, Suffredini AF, et al. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis[J]. Ann Intern Med,1994,120(9): 771-783.
[92] 赵永亮. 特异抗体治疗内毒素血症[J]. 国外医学外科学分册,2002,29(3):132-135.
[93]Moncada S, Higgs A. The L-arginine-nitric oxide pathway[J]. N Eng J Med,1993,329(6):2202-2212.
[94]冉瑞琼,吴裕灯,付华,等. 糖皮质激素对人胃癌细胞肿瘤坏死因子受 体的调节[J]. 中国免疫学杂志,1997,13(2):81-85.
[95]王钧,孙滨,刘林英,等. 地塞米松对内毒素肺损伤绵羊血浆、肺淋巴液神经肽和皮质醇含量的影响[J]. 中国病理生理杂志,1999,15(11):1006-1009.
[96]Haddad EB, Selmon M, Koto H, et al. Ozone induction of cytokine induced neutrophil chemoattractant and nuclear factor-kappa B in rat lung: inhibition by corticosteroids[J]. Febs Lett,1996,379(4):265-268.
[97]常青. 山莨菪碱对内毒素血症大鼠肠系膜血管内皮细胞与白细胞粘附力的影响[J]. 中国病理生理杂志,1995,11(5):471-475.
[98]刘庆增,王金兰. 人参抗内毒素作用研究[J]. 天津医药,1998, 10(4):11-15.
[99]刘媛媛,许昶,吕美德,等. 人参二醇皂甙对内毒素性休克保护作用机制的实验研究[J]. 中国药理学通报,1992,8(1):60-62.
[100]刘媛媛. 人参二醇皂甙对内毒素休克鼠组织过氧化脂质的影响[J]. 白求恩医科大学学报,1990,16(4):342-345.
[101]林桦,李扬,赵雪俭,等. 人参二醇皂甙对休克细胞的保护作用及其机理的实验研究[J]. 白求恩医科大学学报,1992,18(2):86-88.
[102] 赵克森. 休克微循环与细胞流变学变化[J]. 病理生理学报,1980,1(2):47-50.
[103]Kamata H, Manabe T, Kakuta J, et al. Multiple redox regulation of the cellular signaling system linked to AP-1 and NF-kappaB: effects of N-acetylcysteine and H2O2 on the receptor tyrosine kinases, the MAP kinase cascade, and IkappaB kinases[J]. Ann N Y Acad Sci,2002,973:419-422.
[104]Holzheimer RG. The significance of endotoxin release in experimental and clinical sepsis in surgical patients-evidene for antibiotic-induced endotoxin reliease[J]? Infection, 1998,26(8):77-84.
[105]Laflamme N, Rivest S. Toll-like receptor 4: the missing link of the cerebral innate immune response triggered by circulating gram-negative bacterial cell wall components[J]. FESEB,2001, 15(1):155-163.
[106]Ushio S, Namba M, Okura T, et al. Cloning of the cDNA for human IFN-gamma inducing factor, expression in Escherichia coli, and studies on the biological activities of the protein[J]. J Immunol,1996,156(8):4274-4279.
[107]Abraham E. NF-κB activation[J]. Crit Care Med,2000, 28:100-104.
[108]Kim H, Seo JY, Roh KH, et al. Suppression of NF-κB activation and cytokine production by N-acetylcysteine in pancreatic acinar cells[J]. Free Radical Bio & Med,2000,29(7):674-683.
[109]Satoshi K, Masato N, Shunichiro K, et al. Stretch-induced IL-6 secretion from endothelial cells requires NF-κB activation[J]. Bio and Bio Res,2003,308,306-312.
[110]Han SJ, Ko HM, Choi JH, et al. Molecular mechanisms for lipopolysaccharide-induced biphasic activation of nuclear factor-κB[J]. J Bio Chem,2002,277(47):44715-44721.
[111]Bacci A, Verderio C, Pravettoni E, et al. The role of glial cells in synaptic function[J]. Philos Trans Res,1999,
[2]罗海波,鲍行豪. 细菌内毒素研究进展(第一版)[M]. 北京,人民卫生出版社,1983:1-373.
[3]Bayston KF, Cohen J. Bacterial endotoxin and current concepts in the diagnosis and treatment of endotoxamia[J]. J Med Microbiol,1990,31(4):73-83.
[4]Mitov IG, Terziiski DG. Immunoprophyiaxis and immunotherapy of gram-negative sepsis and shock with antibodies to core glycolipids and lipid A of bacterial lipopolysaccha rides[J]. Infection,1991,19(10):383-390.
[5] 邓瑞麟译. 医学微生物学( 第一版)[M]. 北京, 人民卫生出版社,1983:270-272.
[6]邱世翠,曲云,胡涛英,等. 脂多糖致肺血管内巨噬细胞形态和功能的改变[J]. 滨州医学院学报,1992,15(3):211-212.
[7]Birgitt Stein, Petra Frank, Wilhelm S, et al. Endotoxin and cytokines induce direct cardiodepressive effects in mammalian cardiomyocytes via induction of nitric oxide synthase[J]. J Mol Cell Cardiol,1996,28(7):1631–1639.
[8]Torre-Amione G, Kapadia S, Lee J, et al. Tumor necrosis factor-alpha and tumor necrosis factor receptors in the failing human heart[J]. Circulation,1996,93(12):704-711.
[9]Roig E, Orus J, Pare C,et al. Serum interleukin-6 in congestive heart failure secondary to idiopathic dilated cardiomyopathy [J]. Am J Cardiol,1998,82(10):688-690.
[10]Hotchkiss, RS, Swanson PE, Freeman BD, et al. Apoptotic cell death in patients with sepsis, shock and multiple organ dysfunction[J]. Crit Care Med,1999,27(9):1230-1251.
[11]Ayala A, Xu YX, Ayala CA, et al. Increased mucosal B-lymphocyte apoptosis during polymicrobial sepsis is a Fas ligand but not an endotoxin mediated process[J]. Blood,1998, 91(10): 1362-1372.
[12]Alfred Ayala, Joanne L, Patricia S,et al. Pathological aspects of apoptosis in severe sepsis and shock[J]? The International Journal of Biochemistry & Cell Biology,2003,35(6):7-15.
[13]Francoise MA, Luc, DL, Adjuvants[J]. Immunology Today,1993, 14(6):281-293.
[14]李桂杰,朱瑞良,郭宝聚. 细菌内毒素的研究进展[J]. 山东农业大学学报,1998,30(2):193-198.
[15]陈越,金久善. 自由基的生物学效应[J]. 中国兽医杂志,1996, 22(2):48-51.
[16]Remick D. Applied molecular biology of sepsis[J]. J Crit Care,1995,10(4):198-212.
[17]Cowan DB, Poutias DN, Del PJ, et al. CD14-independent activation of cardiomyocyte signal transduction by bacterial endotoxin[J]. Am J Physiol Heart Circ Physiol, 2000, 279(2): 619-629.
[18]Martin TR. Recognition of bacterial endotoxin in the lungs[J]. Am J Respir Cell Mol Biol,2000,23(5):128-132.
[19]Tobias PS, Soldau K, Ulevitch RJ. Isolation of a lipopolysaccharide binding acute phase reactant from rabbit serum[J]. J Exp Med,1986,164(10):777-793.
[20]Heumann D, Roger T. Initial responses to endotoxins and Gram-negative bacteria[J]. Clin Chimica Acta,2002,323(8): 59-72.
[21]李永旺,麻莉,毛宝龄,等. 内毒素诱导的TLR4-MD2信号传导通路[J]. 中国药理学通报,2002,18(5):121-124.
[22]Derek S,J ong S, Edward A. Sepsis:current concepts in intracellular signaling[J]. Int J Biochem,2002,1302(3):1-7.
[23]Leanne P, Siamon C. The function of scavenger receptors expressed by macrophages and their role in the regulation of inflammation[J]. Micro infect,2001,3(1):149-159.
[24]Aderem A, Ulevitch RJ, Kawai T, et al. Toll-like receptors in the induction of the innate immune response[J]. Nature,2000, 406:782-787.
[25]Hemmi H, Takeuchi O, Kawai T, et al. A Toll-like receptor recognizes bacterial DNA[J]. Nature,2000,408:740.
[26]Medzhitov R, Preston P, Janeway CA, et al. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature,1997,388:394-397.
[27]Poltorak A, Ricciardi P, Citterio S, et al. Physical contact between lipopolysaccharide and Toll-like receptor 4 revealed by genetic complementation [J]. Proc Natl Acad Sci,2000,97: 2163.
[28]Ohashi K, Burkart V, Floh S, et al. Heat shock protein 60 is a putative endogenous ligand of the Toll-like receptor 4 complex[J]. J Immunol,2000,164:558-561.
[29]Lien E, Seiiati TJ, Yoshimura A, et al. Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products[J].Chem,1999,274(47):33419-33425.
[30]Yang RB, Mark MR, Gumey AL, et al. Signaling events induced by lipopolysaccharide-activated toll-like receptor 2[J]. J Immumol,1999,163(2):639-643.
[31]Bauer S, Kirschning CJ, Hacker H, et al. Human TLR9 corrfers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci,2001,98(16):9237-9242.
[32]Hemmi H, Takeuchi O, Kawai T, et al. A Toll-like receptor recognizes bacterial DNA[J]. Nature,2000,408:740-745.
[33]Takeuchi O, Kaufmann A,Grote K, et al. Preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-activates immune cells through a Toll-like receptor 2 and MyD88-dependent signaling pathway[J]. J Immunol,2000,164:554-557.
[34]Siebenlist U, Franzoso G, Brown K, et al. Structure, regulation and function of NF-κB[J]. Annu Rev Cell Biol,1994,10:405-455.
[35]Baeuerie PA, Baltimore D. NF-κB: ten years after[J]. Cell,1996,87(1):13-20.
[36] Karin M, Ben-Netiah Y. Phosphorylation meets ubiquitination: the control of NF-κB activity[J]. Annu Rev Immunol,2000, 18:621-663.
[37]Latimer M, Ernst MK, Dunn LL, et al. The N-Terminal domain of IκB masks the nuclear localization signals of p50 and c-Rel homodimers[J]. Mol Cell Bio,1998,18:2640-2649.
[38]Korner M, Rattner A, Mauxion F, et al. A brain-specific transcription activator. Neuron,1989,3:563-572.
[39]Mattson MP, Camandola S. NF-κB in neuronal plasticity and neurodegenerative disorders[J]. J Clin invest,2001, 107(3): 247-254.
[40]Ferret I, Marti E, Lopez E, et al. NF-κB immuno reactivity is observed in association with beta A4 diffuse plaques in patients with Alzheimer's disease.Neuropathol Appl Neurobiol, 1998, 24 (4): 271 -277.
[41]eck S, Lezoualc'h F, Engert S, et al. Insulin-like growth factor-l-mediated neuroprotection against oxidative stress is associated with activation of nuclear factor kappa B. J Biol Chem, 1999, 274(14):9828-9835.
[42]Ginn-Pease ME, Whisler RL. Redox signals and NF-κB activation in T cells . Free Radic Biol Med, 1998, 25(3):346-361.
[43]Ginn-Pease ME, Whisler RL. Redox signals and NF-κB activation in T cells . Free Radic Biol Med, 1998, 25(3):346-361.
[44]Yamamoto Y, Gaynor RB. Therapeutic potential of inhibition of the NF-κB pathway in the treatment of inflammation and cancer[J]. J Clin Invest,2001,107(2):135-142.
[45]Demling RH, Lalonde C. Relationship between lung injury and lung lipid-peroxidation caused by recurrent endotoxin[J]. Am J RevRespir D,1989,139(9):1118-1124.
[46]Sakaguchi S, Kanda N, Hsu CC, et al. Lipid peroxide for-mation and membrane damage in endotoxin-poisoned mice[J]. Microbiol Immunol,1981,25(3):229-244.
[47]Chambers DE, Parks DA, Patterson G, et al.Xanthineoxidaseas a sourse of free radical damage in myocardial ischemia[J]. J Mol Cell Cardiol,1985,17(8):145-154.
[48]Beckman JK. The role of phospholipase: A activity in rat liver microsomal lipidperoxidation[J]. J Biol Chem,1987, 262(4): 1479-1484.
[49]邱海波,陈德昌. 一氧化氮生物作用的两面性与多器官功能障碍综合征[J]. 基础医学与临床,1998,18(1):11-14.
[50]Thiemermann C, Vane JR. Inhibition of nitric oxide synthesis reduces the hypotension induced by bacterial lipopolysacch-aride in the rat[J]. Eur J Pharmacol,1993,182(11):591-595.
[51]Julou-Schaeffer G, Gray G, Fleming, et al.Loss of vascular responsiveness induced by endotoxin involves the L-arginine pathway[J]. Am. J. Physiol.,1991,259(9):H1038-H1043.
[52]Rees DD, Cellek S, Pahner RM, et al. Dexamethasone prevents the induction of nitric oxide synthase and the associated effects on the vascular tone: an insight into endotoxic shock[J]. Biochem Biophys Res Commun,1990,173(11):541-547.
[53]Kilbourn RG, Juburan A, Gross SS, et al. Reversal of endotoxin-mediated shock by N-monomethyl -L-arginine, an
inhibitor of nitric oxide synthesis[J]. Biochem Biophys Res
Commun,1990,172(2):1132-1138.
[54]Kilbourn RG, Gross SS, Jubran A, et al. N-methyl-L-arginine inhibits tumour necrosis factor-induced hypotension: implications for the involvement of nitric oxide[J]. Proc Natl Acad Sci USA,1990,87(7):3629-3632.
[55]Billiar TR, Curran RD, Harbrecht BG, et al. Modulation of nitrogen oxide synthesis in vivo: NG-monomethyl-L-arginine inhibits endotoxin-induced nitrite/nitrate biosynthesis while promoting hepatic damage[J]. J Leukoc Biol,1990,48(12): 565-569.
[56]高洪,陈利平,唐兆新. 内毒素诱生自由基致膜损伤机理的研究进展[J]. 动物医学进展,1999,20(3):1-5.
[57]Lowenstein CJ, Glatt CS, Bredt DS, et al. Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme[J]. Proc Natl Acad sci USA,1992,89(8):6711-6715.
[58]Rockey DC, Chung JJ. Regulation of inducible nitric oxide synthase in hepatic sinusoidal endothelial cells[J]. Am J physiol,1996,271(10):260-267.
[59]Curran RD, Billiar TR, Stuehr DJ, et al. Multiple cytokines are required to induce hepatocyle nitric oxide production and inhibit total protein synthesis[J]. Ann Surg,1990,212(5): 462-469.
[60]Helyar L, Bundschuh DS, Lskin JD. Induction of hepatic Ito cell nitric oxide production after acute endoxoxemia[J]. Hepatology,
1994,20(2):1509-1515.
[61]谢平初,陆家齐,孙大铭,等. 猪内毒素休克外周与内脏微循环灌注的动态变化[J]. 中国危重病急救医学,1999,11(12):718-720.
[62]Palmer RM, Ferrige A, Mocada,S et al. Nitric oxide release accounts for the biological activity of endothelium relaxing factor[J]. Nature,1987,327(12):524-530.
[63]Matute C, Alberdi E, Ibarretxe G, et al. Excitotoxicity in glial cells[J]. Eur J Pharmacol,2002,447:239-246.
[64]Schousboe A. Transport and metabolism of glutamate and GABA in neurons are glial cells[J]. Int Rev Neurobiol,1981,22:1-45.
[65]Matsushita Y, Shima, Nawashiro H, et al. Real-time monitoring of glutamate following fluid percussion brain injury wigh hypoxia in the rat[J]. J Neurotrauma,2000,17(2):143-153.
[66]Fairman WA, Vandenberg RJ, Arriza,JL, et al. Gluamate uptake[J]. Nature,1995,375:599-603.
[67]Ye Z, Sontheimer H. Modulation of glial glutamate transport through cell interactions with the extracellular matrix[J]. Int J Dev Neurosci,2002,20:209-214.
[68]Maragakis NJ, Rothstein JD. Glutamate transporters in neurologic disease. Arch Neurol,2001,58:365-370.
[69]Seal RP, Amara SG. Excitatory amino acid transperters: a family in flux[J]. Annu Rev Pharmacol Toxicol,1999,39:431-456.
[70]Niels C, Danbolt. Glutamate uptake[J]. Prog Neurobiol,2001,65: 100-105.
[71]Schluter K, Figiel M, Rozyczka J, et al. CNS region –specific regulation of glial glutamate transporter expression[J]. Eur J Neurosci,2002,16:836-842.
[72]OShea R. Roles and regulation of glutamate transporters in the central nervous system[J]. Clin Exp Pharmacol Physiol, 2002,29:1018-1023.
[73]Auger C, Attwell D. Fast removal of synaptic glutamate by postsynaptic transporters[J]. Neuron,2000,28:547-558.
[74]Bridges RJ, Kavanaugh MP, Chamberlin AR. A pharmacological review of competitive inhibitors and subatrates of high-affinity, sodium-dependent glutamate transport in the central nervous system[J]. Curr. Pharm Des,1999,5:363-379.
[75]Christopher M, Anderson, Raymond A, et al. Astrocyte glutamate transport: Review of properties, regulation, and physiological functions[J ]. Glia,2000,32:1-14.
[76]Had-Aissouni L, Re DB, Nieoullon A. Importance of astrocytic inactivation of synaptically released glutamate for cell survival in the central nervous system-are astrocytes vulnerable to low intracellular glutamate concentrations[J]? J Physiol Paris,2002,96:317-322.
[77]Kanai Y, Smith CP, Hediger MA. A new family of neurotransmitter transporters: the high-affinity glutamate transporters [J].FASEB J,1993,7(15):1450-1459.
[78]Denis F, Daignan B. Synthesis of glutamine, glycine and 10-formyl tetrahydrofolate is coregulated with purine biosynthesis in Saccharomyces cerevisiae[J].Mol Gen
Genet,1998,259(3):246-255.
[79]Nicholas J, Maragakis, Jeffrey D, et al. Glutamate transporters in neurologic disease[J]. Basic Sci Semin Neurol,2001, 58:365-370.
[80]Volterra A, Trotti D, Tromba C, et al. Glutamate uptake inhibition by oxygen free radicals in rat cortical astrocytes[J]. J Neurosci,1994,14:2924-2932.
[81]Fine SM, Angel RA, Perry SW, et al. Tumor necrosis factor alpha inhibits glutamate uptake by primary human astrocytes[J]. J Biol Chem,1996,271:15303-15306.
[82]Liao SL, Chen CJ. Differential effects of cytokines and redox potential on glutamae uptake in rat cortical glial cultures[J]. Neurosci Lett,2001,299:113-116.
[83]Wang Z, Pekarskaya O, Bencheikh M, et al. Reduced expression of glutamate transporter EAAT2 and impaired glutamate transport in human primary astrocyes exposed to HIV-1 gp 123[J]. Virology,2003,312:60-73.
[84]Pogun S, Dawson V, Kuhar MJ. Nitric oxide inhibits 3H-glutamate transport in synaptosomes[J]. Synapse,1994,18(1):21-26.
[85]Lonart G, Johnson KM. Inhibitory effects of nitric oxide on the uptake of [3H]dopamine and [3H]glutamate by striatal synaptosomes[J]. J Neurochem,1994,63(6):2108-2117.
[86]Trotti D, Aoki M, Pasinelli P. Amyotrophic lateral sclerosis-linked glutamate transporter mutant has impaired glutamate clearance capacity[J]. J Biol Chem,2001,276:576-
582.
[87]Hu SX, Sheng WS, Ehrlich LC, et al. Cytokine effects on glutamate uptake by human astrocytes[J]. Neuroimmunomod-ulation,2000,7:153-159.
[88]Piani D, Frei K, Pfister HW, et al. Glutamate uptake by astrocytes is inhibited by reactive oxygen intermediates but not by other macrophage-derived molecules including cytokines, leukotrienes or platelet-activating factor[J].J Neuroimmunol,1993,48(1):99-104.
[89]全竹富. 抗内毒素核心糖脂抗体:防治革兰氏阴性菌感染的研究现状[J]. 国外医学·外科学分册,1995,22(5):284-288.
[90]Elsbach P, Weiss J, Doerfler M, et al. The bactericidal/ permeability increasing protein of neutrophils is a potent antibacterial and anti-endotoxin agent in vitro and in vivo[J]. Prog Clin Biol Res,1994,388(2):41-54.
[91]Natanson C, Hoffmea WD, Suffredini AF, et al. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis[J]. Ann Intern Med,1994,120(9): 771-783.
[92] 赵永亮. 特异抗体治疗内毒素血症[J]. 国外医学外科学分册,2002,29(3):132-135.
[93]Moncada S, Higgs A. The L-arginine-nitric oxide pathway[J]. N Eng J Med,1993,329(6):2202-2212.
[94]冉瑞琼,吴裕灯,付华,等. 糖皮质激素对人胃癌细胞肿瘤坏死因子受 体的调节[J]. 中国免疫学杂志,1997,13(2):81-85.
[95]王钧,孙滨,刘林英,等. 地塞米松对内毒素肺损伤绵羊血浆、肺淋巴液神经肽和皮质醇含量的影响[J]. 中国病理生理杂志,1999,15(11):1006-1009.
[96]Haddad EB, Selmon M, Koto H, et al. Ozone induction of cytokine induced neutrophil chemoattractant and nuclear factor-kappa B in rat lung: inhibition by corticosteroids[J]. Febs Lett,1996,379(4):265-268.
[97]常青. 山莨菪碱对内毒素血症大鼠肠系膜血管内皮细胞与白细胞粘附力的影响[J]. 中国病理生理杂志,1995,11(5):471-475.
[98]刘庆增,王金兰. 人参抗内毒素作用研究[J]. 天津医药,1998, 10(4):11-15.
[99]刘媛媛,许昶,吕美德,等. 人参二醇皂甙对内毒素性休克保护作用机制的实验研究[J]. 中国药理学通报,1992,8(1):60-62.
[100]刘媛媛. 人参二醇皂甙对内毒素休克鼠组织过氧化脂质的影响[J]. 白求恩医科大学学报,1990,16(4):342-345.
[101]林桦,李扬,赵雪俭,等. 人参二醇皂甙对休克细胞的保护作用及其机理的实验研究[J]. 白求恩医科大学学报,1992,18(2):86-88.
[102] 赵克森. 休克微循环与细胞流变学变化[J]. 病理生理学报,1980,1(2):47-50.
[103]Kamata H, Manabe T, Kakuta J, et al. Multiple redox regulation of the cellular signaling system linked to AP-1 and NF-kappaB: effects of N-acetylcysteine and H2O2 on the receptor tyrosine kinases, the MAP kinase cascade, and IkappaB kinases[J]. Ann N Y Acad Sci,2002,973:419-422.
[104]Holzheimer RG. The significance of endotoxin release in experimental and clinical sepsis in surgical patients-evidene for antibiotic-induced endotoxin reliease[J]? Infection, 1998,26(8):77-84.
[105]Laflamme N, Rivest S. Toll-like receptor 4: the missing link of the cerebral innate immune response triggered by circulating gram-negative bacterial cell wall components[J]. FESEB,2001, 15(1):155-163.
[106]Ushio S, Namba M, Okura T, et al. Cloning of the cDNA for human IFN-gamma inducing factor, expression in Escherichia coli, and studies on the biological activities of the protein[J]. J Immunol,1996,156(8):4274-4279.
[107]Abraham E. NF-κB activation[J]. Crit Care Med,2000, 28:100-104.
[108]Kim H, Seo JY, Roh KH, et al. Suppression of NF-κB activation and cytokine production by N-acetylcysteine in pancreatic acinar cells[J]. Free Radical Bio & Med,2000,29(7):674-683.
[109]Satoshi K, Masato N, Shunichiro K, et al. Stretch-induced IL-6 secretion from endothelial cells requires NF-κB activation[J]. Bio and Bio Res,2003,308,306-312.
[110]Han SJ, Ko HM, Choi JH, et al. Molecular mechanisms for lipopolysaccharide-induced biphasic activation of nuclear factor-κB[J]. J Bio Chem,2002,277(47):44715-44721.
[111]Bacci A, Verderio C, Pravettoni E, et al. The role of glial cells in synaptic function[J]. Philos Trans Res,1999,