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Objective To explore the role of MAPK and NF-?B in the increase of Toll-like receptor 4(TLR4) expression induced by lipopolysaccharide(LPS) in pulmonary micro-vascular endothelial cells(PMVECs) of mouse.Methods PMVECs were isolated,cultured and exposed to the different concentration LPS at(2 h),and(100 ?g/L) LPS in different time.The concentration of TNF-? in culture supernatant was detected using ELISA.(Using) PD98059 inhibitor for ERK,SB203580 inhibitor for P38 MAPK and PDTC inhibitor for NF-?B pretreated(PMVECs).RT-PCR was used to detect the levels of TLR4 and TNF-? mRNA.Western-blot was used to detect the protein expression of nuclear factor-kappa B(NF-?B) P65 subunit and ERK / P38 MAPK in nuclear extract after LPS exposure for 6 h respectively.Results With the treatment of 50～(500 ng /mL) LPS for(2 h) or(100 ?g /L)(LPS exposure) in different time,the concentration of TNF-?in culture supernatant of PMVECs was increased by a time and dosage dependent style(P
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@#ObjectiveTo investigate the effect of α-Zearalanol(α-ZAL) on lipometabolism and hemorheology in ovariectomized(OVX) hyperlipidemia rabbits.Methods44 adult virgin female rabbits were divided into 5 groups,group A: normal control;group B: sham+CHO;group C: OVX+CHO;group D: OVX+CHO+17βE_2;group E: OVX+CHO+α-ZAL.Cholesterol(CHO) was fed to rabbits for 12 weeks.Before and after feeding CHO,the serum lipid(TC,TG,LDL-C,HDL-C) were measured;Blood viscosity,plasma viscosity,aggregation index of RBC(AIRC) and fibrinogen were also assayed respectively.ResultsThe serum levels of TC,TG and LDL-C in group B and E were significantly decreased compared with those in group C(P<0.05);the level of blood viscosity,plasma viscosity and AIRC platelet aggregation rate in group D and E were also significantly decreased compared with those in group C(P<0.05).Conclusionα-ZAL can improve vascular function through the adjustment of lipometabolism and hemorheology.
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BACKGROUND:Lung injury can be induced after injection of lipopolysaccharide(LPS) into the vein of aging rats,and with the development of lung injury,the kidney function can be influenced.Ginkgo biloba extract(GBE) has some effects such as clearing the free radicals,improving hemorrheology,protecting the vascular endothelial cells and so on. OBJECTIVE:To investigate whether renal function damage is induced by acute lung injury(ALI) in aging rats so as to support the hypothesis that lung is the start-up organ in multiple organ dysfunction syndrome in the elderly and the protective effect of GBE on it. DESIGN:Randomized controlled experimental trial based on the experimental animals. SETTING:Laboratory of Department of Pathophysiology,Institute of Basic Medical Science,Peking Union Medical College. MATERIALS:The study has been completed from May 2001 to January 2003 in the Laboratory of Department of Pathophysiology,Institute of Basic Medical Science,Peking Union Medical College.Thirty-six Wistar male rats were involved. INTERVENTIONS:For reproducing the mimic aging rats models,the rats were injected intraperitoneally,D-gal 50 mg/kg,once a day,for 6weeks totally,and randomly divided into 3 groups, namely: ①control group(saline ,intravenous injection);② LPS group( LPS,5 mg/kg intravenous injection) ③ GBE+LPS group (GBE was used 7 days prior to LPS,31 mg/kg ,once a day).Samples(blood,lung and kidney tissue) were collected at 2 or 6 hours after LPS or saline administration. MAIN OUTCOME MEASURES:Contents of creatinine(Cr),blood urea nitrogen (BUN),lactic acid (LA),NO (its metabolite is NO2-/NO3-) and methane dicarboxylic aldehyde(MDA);and activities of glutathione peroxi-dase(GSH-Px) and Na+-K+-ATPase were measured. RESULTS:Compared with control group,there was obviously ALI at 2or 6 hours after LPS administration.Cr [(94.7±10.3) μmol/L]and BUN [(l1.4 1.9)mmol/L]contents in blood were increased significantly until 6 hours after LPS administration.In LPS group,MDA[(22.5 2.6) nmol/L]and NO2-/NO3-[(25.8 2.9) μmol/g] contents in blood and in lung tissue were all increased significantly at 2 hours.But GSH-PX [(355.145.0)μkat/g],Na+-K+-ATPase [(886.3 97.2) nkat/g] activities in lung tissue were decreased significantly at 2 hours after LPS administration.The above changes lasted for 6 hours.However,significant change did not occur until 6 hours after LPS administration in renal tissue.All the above changes were markedly attenuated by pretreatment of GBE. CONCLUSION:ALI was caused by LPS intravenous injection in aging rats.Renal function damage could be induced by ALI in aging rats.GBE showed a protective effect on ALI and renal function damage in this animal model.
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@#ObjectiveTo investigate the effect of Jiuqiang Naoliqing(JNQ)-containing serum on secretion of nitric oxide(NO) and endothelin-1(ET-1) in human umbilical vein endothelial cells(HUVECs), in order to explore the effect of JNQ on regulation of vascular active factors in HUVECs.MethodsThe HUVECs were explored to different volume fractions of JNQ containing serum after being isolated and cultured. The levels of cultured medium of NO and ET-1 were measured.ResultsThe cultured medium content of NO and ET-1 in different volume fractions of JNQ containing serum was significantly increased compared with normal serum control (P<0.05), while the ratio of NO to ET-1 were increased in comparison with normal control (P<0.05).Conclusion JNQ can be promoted secretion of NO and ET-1 in HUVECs, preserving endothelial function, and maintaining the balance of NO/E-1.
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Objective To investigate whether gingko biloba extract (GBE) have protective effect on acute lung injury(ALI)induced by Lipopolysaccharide ( LPS) in aging rats and renal function damage induced by ALI. Methods Male aging Wistar rats(36) were used in the study. LPS group (LPS, iv route, 5mg/kg body weight) and GBE+LPS group (GBE was given starting from 7 days before experiment, once a day oral administration). The blood, lung and kidney samples were collected at 2 or 6 hours after LPS or saline administration. Results Compared with the aging control, ALI was obviously observed in LPS group, blood creatinine and urea nitrogen were increased significantly from (68.7? 6.9) mol/L and (5.9?0.6) mmol/L to (94.7?10.3) mol/L and (11.4?1.9) mmol/L, respectively (P
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In this study,the relationship between the advanced glycation end products(AGEs) and the expressions of receptor for AGEs(RAGE),intercellular cell adhesion molecule-1(ICAM-1) was investigated.The diabetic rat model was reconstructed and the fluorescence method,RT-PCR and in-situ hybridization techniques were used to detect AGEs content and the expressions of RAGE and ICAM-1 gene in the aorta and cardiac tissues.The results showed that AGEs content in aortic and cardiac tissues increased(P＜0.01) in diabetic rats; The expressions of RAGE and ICAM-1 enhanced (P＜0.05～0.01) and were positively correlated with the quantity of AGEs accumulation(P＜0.01) in the aorta and cardiac tissue.These parameters change in the diabetic rats can be improved with aminoglumine(AG) treatment.Suggesting that AGEs might induce RAGE and ICAM-1 expression.It's postulated that AGEs binding to RAGE play an important role to result in diabetic endothelial cells dysfunction and lesion.
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AIM: To investigate the effects of a Chinese herb compound prescription, Wen-Jin-Tong(WJT), on the deposition of advanced glycation end products (AGEs) ,the expression of receptor for AGEs(RAGE)and ICAM-1 in cardiovascular tissues of diabetic rats. METHODS: The diabetic rat model was induced by injection of streptozotocin. The quantity of AGEs accumulation, the expressions of RAGE and ICAM-1 in aorta and cardiac tissues were detected using fluorescence method, RT-PCR and in situ hybridization techniques. RESULTS: AGEs accumulation in aorta and cardiac tissues of rats treated with Wen-Jin-Tong was lower than that of diabetic model rats(P
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AIM:The objective of this study was to observe the effects of AP-1 element on endothelin-1(EDN1)gene transcription in homocysteine(Hcy)-induced human umbilical vein endothelial cells(HUVECs).METHODS:The redox level in Hcy-induced HUVECs was determined by using the fluorescent product DCF.The influences of Hcy on expressions of EDN1 mRNA and protein of c-jun/AP-1 in HUVECs were measured by the methods of RT-PCR and Western blotting,respectively.The EDN1 secretion level of HUVECs induced by Hcy was determined by enzymatic immunoassay.The transient transfection assay was performed and luciferase activity of transcriptional factor AP-1 reporter gene induced by Hcy was detected.RESULTS:The Hcy significantly increased EDN1 secretion,EDN1 mRNA expression and oxidative stress in HUVECs.Hcy remarkably induced the expression of pGL3-EDN1-AP-1(115/+135)luciferase reporter gene plasmid.However,basic expression of mutation of pGL3-EDN1-AP-1(-115/+135)luciferase reporter gene plasmid was downregulated markedly in HUVECs induced by Hcy.CONCLUSION:Redox-active and release of ROS are changed by Hcy.Activated AP-1 element plays an important role in EDN1gene transcription in HUVECs induced by Hcy.
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Objective To provide laboratory data and direct experience of inhaling nitric oxide (NO) to treat persistent pulmonary hypertension of neonate.Methods Examined the effect of No on pulmonary arterial pressure of inhaled NO while rabbits were inhaled 44ppm NO,5% O_2 and air respectively in this model and those inspired air.Results Pulmonary hypertension was induced by reducing the fraction of inspired oxygen to 5% for 3 minutes.While 44ppm NO was added,a rapid decrease in pulmonary arterial pressure was observed (P
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AIM: To reveal whether H2O2, a kind of platelet - derived growth factor (PDGF) signaling molecule, involved in the proliferation of phypoxic pulmonary arlmonary artery smooth muscle cells (PASMCs) and expression of transcription factor NF -?B. METHODS: bar confocal scanning microscopy,immunocytochemistry and Western blot analysis were used. RESULTS: PDGF (30 ng/mL) elevated the intracellular H2O2 in normoxic PASMCs, but decreased that of hypoxic PASMCs significantly. When 100?mol/L H2O2 was added, NF - ?B expression were inhib- ited markedly in hypokic PASMCs, while the same concentration H2O2 resulted in increase of NF - ?B expression in control cells. CONCLUSION: H2O2, a mediator molecule of PDGF signal transduction, act as an inhibitor in hy- poxic PASMCs which is qUiet different from that in normoxic PASMCs. It may effect on the expression of transcription factor NF -?B. NF - ?B induce various gene expression that involving in the proliferation of vascular smooth muscle cells. One mechanism by which PDGF - induced hypoxic PASMC proliferation might be through PDGF alleviating H2O2 inhibition.
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AIM:To investigate whether renal function damage could be induced by acute lung injury(ALI)in aging rats. METHODS: 40 Wistar male rats were used for reproducing aging animal model. Aging rats were randomly divided into aging control; ALI group (Lipopolysaccharide, LPS, 5 mg/kg body weight, iv) and LPS group (same dosage LPS, intraventricle of left heart injection). The samples (blood, lung and kidney ) were collected at 2 or 6 hours after LPS or normal saline administration. RESULTS:Compared with aging control, in ALI group, creatinine (Cr) and urea nitrogen (BUN) contents in blood were increased significantly (all P