Effect of Morphine and Naloxone on Release of the Excitatory Amino Acids of Spinal Astrocytes Induced by TNF-α

The effect of morphine and naloxone on release of the excitatory amino acids (EAAs) of spinal astrocytes induced by TNF-α was studied. Astrocytes were purified from 2- to 3-day old SD rats and divided into 8 groups: group 1 (without any stimulatants); group 2 (10 ng/ml TNF-α); group3 (10 ng/ml TNF-α+0.5 μmol/L morphine); group 4 (10 ng/ml TNF-α+1.0 μmol/L morphine); group 5 (10 ng/ml TNF-α+2.0 μmol/L morphine); group 6 (10 ng/ml TNF-α+0.5 μmol/L naloxone); group 7 (10 ng/ml TNF-α+1.0 μmol/L naloxone); group 8 (10 ng/ml TNF-α +2.0 μmol/L naloxone). In group 2, 3, 4 and 5, 0, 0.5, 1.0 or 2.0 μmol/L morphine was added to the cells cultured with serum-free Neurobasal/B27 medium containing 10 ng/ml TNF-α respectively, while in group 6, 7 and 8, 0.5, 1.0 or 2.0 μmol/L naloxone was added respectively to the cells cultured with serum-free Neurobasal/B27 medium containing 10 ng/ml TNF-α. After 30 min incubation, high-pressure liquid chromatography (HPLC) was used to measure the levels of EAAs in all cultured cells. The results showed the level of EAAs in group 2 was significant higher than in group 1 (P<0.01). As compared with group 2, the levels of EAAs in group 3, 4 and 5 were decreased with the difference being significant between group 5 and group 2 (P<0.01) or between group 4 and group 2 (P<0.05). The levels of EAAs in group 6, 7 and group 8 was significantly lower than in group 2 (P<0.05 orP<0.01). It was concluded that TNF-α could promote the release of glutamate and aspartate from astrocytes, and morphine and naloxone might reduce the release of EAAs in cultured spinal astrocytes induced by TNF-α.     

Expression of macrophage inflammatory protein 1α in the endothelial cells exposed to diamide

Summary In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1α (MIP-1α), the expression of MIP-1α protein in the cells was detected by cell enzyme-linked immunosorbent assay (ELISA) and that of MIP-1α mRNA was determined by cell in situ hybridization and nuclease S1 protection assay after the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activity of MIP-1α was tested by micropore filter method using modified Boyden chambers. Cell ELISA showed that the expression of MIP-1α protein in endothelial cells exposed to 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the control cells, which was statistically significant by analysis of variance. In situ hybridization revealed that the mRNA expression of ECs treated with 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.3-fold, 3.0-fold and 1.7-fold as much as that in the control group, which had statistical significance (F=188. 93,P<0.01). The mRNA expression in 5 μmol/L dimide treated ECs, measured by nuclease S1 protection assay, was 3.4-fold as much as that in the control group (t=8.70,P<0.05). Chemotactic response (99.50±4.31 μm) to the culture medium conditioned by 5 μmol/L diamide treated ECs, which was stronger than that (66.47±3.25 μm) conditioned by the ECs (F=404.31,P<0.05), was significantly decreased (F=192.25,P<0.05) after adding MIP-1α antibody. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce high level of MIP-1α, and might play an important role in atherogenesis by promoting the migration of peripheral blood monocytes into arterial intima. YANG Limin, female, born in 1973, M. D., Ph.D. This project was supported by a grant from National Natural Sciences Foundation of China (No. 39730220).     

Effects of propofol on the mRNA expression of toll-like receptor 4 in BV-2 cells during mimic ischemia-reperfusion injury <Emphasis Type="Italic">in vitro</Emphasis>

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 （TLR4） in BV-2 cells during mimic ischemia-reperfusion （I/R） injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random： control group （group C）, ischemia/reperfusion group （group I/R）, low-dose propofol （25 μmol/L） intervention group （group PF25） and high-dose propofol （100 μmol/L） intervention group （group PF100）. The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C （P〈0.01）. And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C （P〈0.01）. After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R （P〈0.01）. And the decrease in those indicators was more significant in group PF100 than in group PF25 （P〈0.01）. It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Effect of IL-Iβ and TNF-α on the expression of monocyte chemotactic protein-1 in endometriotic cells

Summary To investigate the clinical significance of monocyte chemotactic protein-1 (MCP-1) produced by endometriotic tissues, the endometriotic tissues were taken from 15 patients with endometriosis. MCP-1 mRNA and MCP-1 protein were determined by dot blot analysis and enzyme linked immunosorbent assay (ELISA) in endometriotic cells cultured with or without interleukin-1β (IL-lβ, 2 μg/L), tumor necrosis factor-α (TNF-α, 20 g/L). After exposure to IL-1β or TNF-α, the expression of MCP-1 mRNA in the endometriotic cells (8.635 ±0.826, 7.031 ±0.970, respectively) were significantly higher than that in the control group (4.482±0.435, P<0.05); The expression of MCP-1 protein in IL-lβ and TNF-α group was 4.52±0.09 μg/L,2.87±0.27 μg/L, respectively, which were significantly higher than 1.74±0.16 μg/L in control (P<0.01). The results suggested that IL-l\ and TNF-α could up-regulate the expression of MCP-1 in endometriotic cells, which might be related to the development of endometriosis.     

Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide

Objective:To evaluate the effect of Jianpi Huoxue decoction(健脾活血方,JHD)-containing serum on tumor necrosis factor-α(TNF-α) secretion and endotoxin receptor gene expression in RAW264.7 cells induced by lipopolysaccharide(LPS).Methods:The cytotoxicity of blank-control serum and JHD-containing serum at different concentrations were evaluated through the lactate dehydrogenase(LDH) assay in RAW264.7 cells.RAW264.7 cells were divided into six groups:5%blank-control serum group(C1,n=3),5%blank-control serum plus...     

Effect of Shengmai injection (生脉注射液) on vascular endothelial and heart functions in patients with coronary heart disease complicated with diabetes mellitus

Objective： To study the effect of Shengmai injection （生脉注射液, SMI） on vascular endothelial and heart functions in coronary heart disease patients complicated with diabetes mellitus （CHD-DM）. Methods： One hundred and twenty patients with CHD-DM, their diagnosis confirmed by coronary arteriography, were equally randomized into a control group treated with conventional treatment and a treated group treated with conventional treatment plus SMI. The changes in blood levels of nitric oxide （NO）, endothelin-1 （ET-1） and angiotensin Ⅱ （Ang Ⅱ）, as well as endothelium-dependent vascular dilating function and heart function in the patients were observed before treatment and after the 3-week treatment. Results： After being treated with SMI for 3 weeks, in the treated group, blood level of NO was raised significantly from 69.8±33.1 μmol/L to 120.1±50.8μmol/L, and ET-1 was lowered from 70.1±32.1 ng/L to 46.2±21.3 ng/L, respectively （P〈0.01）; that of Ang Ⅱ was lowered from 81.3±24.3 ng/L to 50.2±27.3 ng/L （P〈0.01）; brachial arterial post-congestion blood flow increasing rate was raised from 389.4±26.3% to 459.3±27.8% （P〈0.01）; and the improvement in heart function as seen through the ejection fraction （EF） was increased from 44±5% to 68±6% （P〈0.01）, all the changes being more significant than those in the control group （all P〈0.01）. Conclusion： SMI can improve not only the endothelial function in CHD-DM patients, but also heart contraction significantly.     

Effect of Naotaifang extract on changes of coagulation in human umbilical cord vein endothelial cell and fibrinolysis function induced by recombinant human Tumor Necrosis Factor-α

Objective: To observe the changes of coagulation and fibrinolysis function in human umbilical cord vein endothelial cells (HUVEC) induced by recombinant human tumor necrosis factor-α (rhTNF-α) and the effect of Naotaifang extract (NTE) on it.Methods: Cultured HUVEC is randomly divided into six groups: Control group, NTE control group (only 2 g/L NTE), rhTNF-α group (100 μg/ L rhTNF-α), and low-dosage, middle-dosage and high-dosage NTE group (100 μg/L rhTNF-α and 0. 67 g/L, 2 g/L, 6 g/L NTE). The coagulation activity of frozen-dissolved HUVEC, von Willebrand factor (vWF) content in the conditioned medium and tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAD activity were to be detected after 24 hrs.Results: Compared with the control group, PAI activity were enhanced, vWF release markedly increased in conditioned medium of TNF-α group (P < 0.01) and the frozen-dissolved HUVEC markedly shortens the rabbit plasma prothrombin time, and the above changes could be significantly inhibited by the 3 dosages of NTE (P < 0.05,P < 0. 01).Conclusion: NTE is effective in inhibiting the coagulation activity of the HUVEC non-stimulated or stimulated by rhTNF-α to enhance the vWF release, and to adjust fibrinolytic function, and mainly to inhibit the PAI activity. The item is supported by Natural Science Foundation of Hunan Province (No. 98JJY 2027) and Young-middle Aged Science and Technological Fund of Hunan Province (NO. 00JZYZ145)     

Effects of Propyl Gallate on adhesion of polymorphonuclear leukocytes to human endothelial cells induced by tumor necrosis factor alpha

Objective:To investigate the effects of Propyl Gallate(PrG) on cellular adhesion between human umbilical vein endothelial cells(HUVEC) and polymorphonuclear leukocytes(PMN) as well as the expression of intercellular adhesion molecule-1(ICAM-1,CD54) and E-selectin(CD62E) on the VEC surface.Methods: A human VEC inflammation model was induced by tumor necrosis factor alpha(TNF-α).VECs were preincubated with varying concentrations of PrG(0.001-5 mmol/L) or 1‰DMSO(v:v) or 10 mmol/L acetylsalicylic acid(ASA) f...     

Effect of Early Hemofiltration on Pro- and Anti-inflammatory Responses and Multiple Organ Failure in Severe Acute Pancreatitis

Severeacute pancreatitis (SAP )progressesrapidlywithacomplicatedoutcomeandhighmortali ty .Themultipleorgandysfunctionsyndrome(MODS)inducedbytheoverreleaseofcytokineswhichisthemaincauseofpatients’deathsintheear lystage[1] .Hemofiltration (HF)isoneoftheblo…