白头翁汤及其主要成分对LPS诱导内皮细胞分泌NO、E-selectin和IL-8的影响

拟探讨白头翁汤及其主要成分对细菌内毒素(LPS)导致的机体炎症等病理损伤的治疗作用.培养内皮细胞至单层融合状态,加入内毒素刺激,3h后加入高、中、低3种浓度的8种药物,继续培养21 h后收集细胞上清液,用ELISA方法测定NO、E-selectin和IL-8的含量.结果显示,白头翁汤高剂量组、白头翁素高剂量组和小檗碱高剂量组的NO含量极显著低于LPS对照组(P＜0.01),白头翁汤中剂量组、白头翁素中剂量组、小檗碱中剂量组和秦皮乙素高剂量组显著低于LPS对照组(P＜0.05);白头翁汤高剂量组和药根碱高剂量组的E-selectin含量极显著低于LPS对照组(P＜0.01),白头翁皂苷B4高剂量组、小檗碱高剂量组和药根碱中剂量组显著低于LPS对照组(P＜0.05);秦皮乙素高剂量组的IL-8含量极显著低于LPS对照组(P＜0.01),白头翁汤高剂量组、秦皮甲素高剂量组和中剂量组、秦皮乙素中剂量组显著低于LPS对照组(P＜0.05).结果提示白头翁汤及其主要成分具有明显的抗细菌内毒素作用,可在由NO、E-selectin和IL-8介导的机体炎症等病理损伤过程中发挥治疗作用.     

秦公散对LPS活化的内皮细胞表达黏附分子和分泌一氧化氮的影响

为阐明秦公散的抗炎作用机制,本试验采用流式细胞术检测了LPS刺激的内皮细胞黏附分子E-选择素和ICAM-1的表达,用一氧化氮(NO)测定试剂盒检测LPS刺激的内皮细胞培养上清液中NO的含量。结果表明,LPS刺激的内皮细胞可显著提高E-选择素和ICAM-1的表达(P0.01),显著提高培养上清液中NO的含量(P0.05);用秦公散及其含药血清处理后则明显抑制LPS诱导的E-选择素、ICAM-1的表达和NO分泌(P0.05)。表明秦公散可通过抑制黏附分子的表达及NO分泌量发挥抗炎作用。     

Association of serum soluble E-selectin concentrations with insulin resistance in essential hypertension patients

AIM: To investigate the relationship between serum soluble E-selectin (sE-selectin) and insulin resistance, serum uric acid, serum lipid in essential hypertension patients. METHODS: Fasting serum sE-selectin concentration, plasma glucose, serum insulin, serum uric acid, total cholesterol, triglycerides, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol were determined in 186 patients with essential hypertension (75 males, 111 females). Homeostasis model assessment was applied to assess the status of insulin resistance (HOMA-IR). RESULTS: Based on the HOMA-IR, the essential hypertension patients were divided into insulin-sensitive individuals (IS) and insulin resistant subjects (IR). The serum sE-selectin concentration was significantly higher in male group [(50.1±17.8)g/L]than in female group [(40.6±16.6)g/L](P<0.01). No difference between IR group [(51.6±16.8)g/L]and IS group [(48.5±18.8)g/L] in male, and significantly higher in IR group [(45.1±18.0)g/L]than in IS group [(36.0±13.7)g/L](P<0.01) in female were observed. A stepwise multiple linear regression analysis showed that HOMA-IR was an independent predictor of serum sE-selectin concentrations in female group and not in male group, and both serum uric acid and serum lipid were not independent predictors of serum sE-selectin concentrations. CONCLUSION: Serum sE-selectin concentrations were directly related to insulin resistance in females with essential hypertension and not in males with essential hypertension. Both serum uric acid and serum lipid were not directly related to serum sE-selectin concentrations.     

Inhibitory effects of Angelicae sinensis and sodium ferulate on acute inflammatory liver injury and expression of ICAM-1 and E-selectin in mice

AIM: To explore the effects of Angelicae sinensis preparation and sodium ferulate on inflammatory liver injury induced by lipopolysaccharide (LPS) and their molecule mechanism. METHODS: ICR mice were divided into five groups: Angelicae sinensis group, sodium ferulate group, dexamethasone group, inflammation control group and normal group. The model of inflammatory injury in mice was set up by tail vein injection with the bacillus calmette-guerin (BCG) and LPS respectively prior and posterior to administration of those tested drugs. The tested drugs (the preparations of Angelica sinensis, sodium ferulate and dexamethasone) and normal saline were given respectively to the corresponding group. The pathological observation of the liver tissues in mice was made for the intensity of inflammatory liver injuries. The immunohistochemical detection and comparison were performed for the expression of ICAM-1 and E-selectin proteins in the liver tissues in those mice. RESULTS: The intensities of liver inflammatory injuries in mice from drug-treated groups were obviously lighter than that from the inflammation control group (P<0.01). The expressions of ICAM-1 and E-selectin proteins in the liver tissues of mice from the inflammation control group were not only significantly higher than that in normal control, but also obviously higher than that from drug-treated groups (P<0.01). CONCLUSIONS: Angelica sinensis and sodium ferulate alleviate inflammatory liver injury induced by injection of LPS. Their suppressive effects on inflammatory liver injury may be associated with the decrease in the expression of ICAM-1 and E-selectin proteins.     

六种中药生物碱对LPS作用下猪血管内皮细胞分泌细胞因子的影响

通过研究6种中药生物碱对细菌内毒素（LPS）作用下猪血管内皮细胞分泌细胞因子血栓素B2（TXB2）,内皮素-1（ET-1）和E-选择素（E-selectin）的影响,拟探讨生物碱抗LPS的药理作用机制。培养猪血管内皮细胞至单层融合状态,加入LPS刺激,3 h后加入高、中、低3种浓度的6种中药生物碱青藤碱、防己诺林碱、水苏碱、川芎嗪、苦参碱和吴茱萸碱,继续培养21 h后收集细胞上清液,用ELISA测定TXB2,ET-1和E-selectin的含量。结果,水苏碱高剂量组和川芎嗪高剂量组的TXB2含量极显著低于LPS对照组,川芎嗪中剂量组和吴茱萸碱高剂量组显著低于LPS对照组;苦参碱高剂量组的ET-1含量极显著低于LPS对照组,青藤碱高剂量组显著低于LPS对照组;防己诺林碱高剂量组和吴茱萸碱中剂量组的E-selectin含量显著低于LPS对照组。结果提示六种中药生物碱具有明显的抗细菌内毒素作用,可在由TXB2,ET-1和E-selectin介导的机体凝血、休克和炎症等病理损伤过程中发挥治疗作用。     

Effects of ambroxol combined with low-dose heparin on production of ICAM-1 and E,P-selectin in acute lung injury

AIM: To study the production of intercellular adhesion molecule-1(ICAM-1), E-selectin and P-selectin in serum, lung tissues and bronchoalveolar lavage fluid(BALF)of acute lung injury(ALI) model and to observe the effects of ambroxol combined with low-dose heparin on the changes of the 3 factors above.METHODS: Twenty-four healthy rabbits were randomly divided into 3 groups: normal saline control group (NC), oleic acid injury group (OA), ambroxol+ heparin treatment group (AH). The rabbit ALI model was induced by oleic acid injection through auricular vein. Partial pressure of O₂ in artery(PaO₂) was analyzed.The concentrations of ICAM-1 and E-selectin were detected by ELISA.The apoptosis index(AI) was measured by TUNEL method.The expression of P-selectin was determined by immunohistochemical method.The ultrastructural changes of the lung tissues were observed under electron microscope, and the lung wet/dry ratio(W/D) was calculated.RESULTS: PaO₂ in AH group and OA group was significantly lower (P<0.01) than that in NC group, and PaO₂ in AH group was significantly higher than that in OA group (P<0.01). The concentrations of ICAM-1 and E-selectin in serum, lung tissues and BALF, and AI and W/D in lung tissues in AH group were higher (P<0.05 or P<0.01) than those in NC group, and was lower than those in OA group (P<0.05 or P<0.01). In NC group, no significant change of the above parameters at all time points was observed (P>0.05). In OA group, PaO₂ was significantly decreased (P<0.01) with the pathological process developed, and the concentrations of ICAM-1 and E-selectin were significantly increased. In AH group, PaO₂ was decreased (P<0.05),and the concentrations of ICAM-1 and E-selectin were increased with the process of ALI developed. The P-selectin expression in lung tissues of OA group was distributed mainly in inflammatory cells, capillary endothelial cells and plasma. From low to high levels, the order was NC group < AH group < OA group in the expression of P-selectin. The most obvious apoptosis was observed in OA group. No apoptosis or occasional positive cells were found in NC group. The apoptotic rate in AH group was significantly reduced compared with that in OA group.CONCLUSION: In ALI induced by OA, ICAM-1, E-selectin and P-selectin are significantly increased and are involved in the occurrence and development of ALI. Ambroxol combined with low-dose heparin reduces the levels of ICAM-1, E-selectin and P-selectin, the pulmonary edema and the lung injury, improves pulmonary functions, and plays an important role in the prevention and treatment of acute lung injury.     

Paeonol reduces expression of adhesion molecules in HUVECs induced by hyperlipidemic serum via inhibiting the pathway of NF-κB signaling

AIM: To observe the anti-atherosclerosis effect of paeonal (Pae) on the activation of NF-κB and the expression of cell adhesion molecules in human umbilical vein endothelial cells (HUVECs) induced by hyperlipidemic serum. METHODS: Cultured HUVECs were used as target cells. Hyperlipidemic serum was added to the culture medium to establish the injury mode of HUVECs. Methyl thiazolyl tetrazolium (MTT) method was used to examine the cell viability. The mRNA expression of NF-κB p65 was determined by RT-PCR. The protein levels of IκB-α, intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin were detected by Western blotting. RESULTS: After treated with Pae, the cell viability was increased and the morphological changes of HUVECs injured by hyperlipidemic serum trended to normal. The expression of IκB-α in HUVECs injured by hyperlipidemic serum increased, while the expression of NF-κB p65 mRNA, ICAM-1 and E-selectin protein was decreased. CONCLUSION: The anti-atherosclerosis mechanism of paeonal may be related to the inhibitory effect of the natural compound on the pathway of NF-κB/IκB, thereby reducing the expression of ICAM-1 and E-selectin and attenuating the inflammatory reaction in vascellum.     

Chlorella 11-Peptide Inhibits the Production of Macrophage-Induced Adhesion Molecules and Reduces Endothelin-1 Expression and Endothelial Permeability

The inflammation process in large vessels involves the up-regulation of vascular adhesion molecules such as endothelial cell selectin (E-selectin), intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) which are also known as the markers of atherosclerosis. We have reported that Chlorella 11-peptide exhibited effective anti-inflammatory effects. This peptide with an amino sequence Val-Glu-Cys-Tyr-Gly-Pro-Asn-Arg-Pro-Gln-Phe was further examined for its potential in preventing atherosclerosis in this study. In particular, the roles of Chlorella 11-peptide in lowering the production of vascular adhesion molecules, monocyte chemoattractant protein (MCP-1) and expression of endothelin-1 (ET-1) from endothelia (SVEC4-10 cells) were studied. The production of E-selectin, ICAM-1, VCAM-1 and MCP-1 in SVEC4-10 cells was measured with ELISA. The mRNA expression of ET-1 was analyzed by RT-PCR and agarose gel. Results showed that Chlorella 11-peptide significantly suppressed the levels of E-selectin, ICAM, VCAM, MCP-1 as well as ET-1 gene expression. The inhibition of ICAM-1 and VCAM-1 production by Chlorella 11-peptide was reversed in the presence of protein kinase A inhibitor (H89) which suggests that the cAMP pathway was involved in the inhibitory cause of the peptide. In addition, this peptide was shown to reduce the extent of increased intercellular permeability induced by combination of 50% of lipopolysaccharide (LPS)-activated RAW 264.7 cells medium and 50% normal SEVC cell culture medium (referred to as 50% RAW-conditioned medium). These data demonstrate that Chlorella 11-peptide is a promising biomolecule in preventing chronic inflammatory-related vascular diseases.