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Objective To investigate the effects of different doses of ulinastatin on severe thermal injuryinduced myocardial damage in rats. Methods One hundred and eighty female SD rats weighing 180-220 g were usedin this study. Thirty percent of the total body surface (TBS) was shaved chemically with 20% sodium sulphate and then exposed to 92 ℃ water for 18 s. The animals with third degree thermal injury involving 30% of the TBS were randomly divided into 3 groups (n = 60 each): group Ⅰ thermal injury (group TI); group Ⅱ and Ⅲ received intraperitoneal ulinastatin 40 000 and 80 000 U/kg respectively immediately after thermal injury (group U1 , U2). The TI group received equal volume of normal saline IP instead of ulinastatin. Blood samples were taken from abdominal aorta before (baseline) and at 1, 3, 6, 12 h after thermal injury for determination of serum concentrations of cTnI, IL-1β, IL-6, IL-10 and TNF-α (10 samples at each time points). Ten animals were sacrificed at each time point after blood sampling. The myocardial specimens were obtained for microscopic examination and measurement of MDA content and SOD activity. Results Compared with group TI, the serum concentrations of cTnI, IL-1β, IL-6 and TNF-α and MDA content in myocardium were significantly decreased and the myocardial SOD activity was significantly increased in group U1 , while in group U2 the senum concentrations of cTnI, IL-Iβ, IL-6 and TNF-α and myocardial MDA content were significantly increased and the myocardial SOD activity was significantly decreased. There was no siginificant difference in the serum concentrations of IL-10 among the three groups. Microscopic examination showed that myocardial damage was accentuated in group U2 as compared with group U1. Conclusion Ulinastatin 40 000 U/kg can ameliorate severe thermal injury-induced myocardial injury through inhibition of inflammatory response and lipid peroxidation response, whereas ulinastatin 80 000 U/kg accentuates myocardial damage.
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Objective To study the protective role of pre-resolving mediator lipoxin A4(LXA4) in the NA+ -K+-ATPase in alveolar type Ⅱ (AT Ⅱ ) epithelial cells of rats exposed to lipopolysaccharide (LPS). Method The AT Ⅱ cells were isolated and purified, and divided randomly into control group (PBS), vehiculum (alcohol 0.7 μL/mL) group, LPS (1 μg/mL) group, LXA4(1/10 mol/mL) group and LPS (1 μg/mL LPS) + LXA4(1/10 mol/mL) group. After exposure to LPS and/or LXA4 for4 hours, NA+-K+ -ATPase and β1-subunits mRNA in AT Ⅱ epithelial cells were detected by using RT-PCR, and ATP, ADP, AMP, total adenine nucleotides (TAN) and energy charge (EC) were measured by using high performance liquid chromatography (HPLC), and then the activities of Na+-K+-ATPase were calculated accordingly. Results The NA+-K+-ATPase α-subunit and β-subunit mRNA were significantly decreased in LPS group ( P ＜ 0.05 vs. control group). However, the expressions of NA+ -K+-ATPase mRNA were significantly enhanced by application of LXA4 to AT Ⅱ epithelial cells exposed to LPS (P ＜0.05 vs. LPS group). The activities of NA+ -K+ -ATPase were enhanced in LPS group (P ＜0.05 vs. control group). Compared with control group and LPS group, the activities of NA+-K+-ATpase in LPS + LXA4 group were significantly increased (P ＜0.01 vs. control group; P ＜0.05 vs. LPS group). The EC of AT Ⅱ epithelial cells were higher in LPS group ( P ＜ 0.01 vs. control group). There were no significant differences in EC between control group and LPS + LXA4group(P ＞0.05). Conclusions The pro-resolving mediator LXA4 can enhance the expressions of NA + -K + -ATPase α-subunit and β-subunit mRNA, and the activities of NA + -K + -ATPase in AT Ⅱ epithelial cells or rats exposed to LPS, and ca also balance the metabolism of AT Ⅱ epithelial cells. These findings suggest that LXA4 plays an important role in lung edema clearance in lung injury induced by endotoxin, and the role is likely associated with the enhancement of the expressions of Na+ -K+ -AT-Pase α-subunit and β-subunit, and the activities of Na+ -K* -ATPase, maintaining the balance of metabolism of AT Ⅱ epithelial cells.
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Objective To study the effects of Lipoxins A4(LXA4)on the expressions of aquaporin(AQP)1,3,5 in type Ⅱ pneumonocytes(ATⅡ)of rat treated with lipopolysaccharide(LPS).Method One pathogenfree male Spree Dawley(SD)rat every time.weighing 200～250 g,were used for the study.The typeⅡpenumonocytes of rats were isolated and purified,and the changes of cellular ultrastructure were observed by electron microscope in order to get the purity quotien＞90%.The type Ⅱ pneumonocytes were divided randomly into five groups,namely,vebicukun group(alcohol 0.7μL/mL),control group,LXA4 group(1×10-7mol/mL),endotoxin group(LPS 1μg/mL)and LXA4+LPS group(LXA4 1×10-7mol/mL,LPS 1μg/mL).AQP-1,3,5 mRNA of in the typeⅡpenumonocytes were assayed by using reversal transcription poly chain reaction(RT-PCR),and the expressions of AQP-1,3,5 protein were detected by using.immunohistochemistry(IHC).One each specimen,these tests were repeated for six times.ANOVA was used for statistical analysis.Results RT-PCR and IHC showed that when AT Ⅱ treated with 1 μg/mL LPS for 4 hours,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were significantly decreased in LPS group compared with control group(P＜0.01).However,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein after application of LXA4 significandy increased in LPS+LXA4 group in comparison with LPS group(LPS+LXA4,AQP1:0.647±0.132,AQP3:0.900±0.856,AQP5:0.879±0.058;LPS,AQP1:0.297±0.133,AQP3:0.512±0.113,AQP5:0.647±0.110;P＜0.01).The AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were aignificandy increased in LXA4 group in comparison with control group(LXA4,AQP1:0.539±0.142,AQP3:0.818 4-0.176,AQP5:0.841±0.066;Blank Control,AQP1:0.518±0.139;AQP3:0.138±0.136,AQP5:0.766±0.066;P＜0.01).Conclusions AQP-1,3,5 exist in typeⅡpenumonoeyte of rata,and the LXA4 can up-regulate the mRNA and protein expressions of AQP-1,3,5 in Type Ⅱ penumonocytes of rats treated with LPS.
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Objective To assess whether propofol call induce stable psychic dependence in the rats by self-administration experiment. Methods Twenty-four male SD rats 14 weeks old weighing 240一270 mg were studied. Anesthesia was performed with intraperitoneal injection of 3%sodium pentoharbitsl 40 ms/kg and atropine 03 mg/kg.A catheter wag inserted into the right external jugular vein. Penicillin(100 000 U)0.2 ml wag injected through the external jugular vein for anti-infection and heparin sodium(50U/ml)0.1 ml for anticoagulation. The self-administration experiment of 14 days was started after the 7 days of recovery. All the rats were randomly divided into 4 groups(n=6 each):contontrol group(C),propofol 0.56 mg/kg/l group(P1),propofol 1.00 mg/kg group(P2)and pmpofol 1.70 ms/kg group(P3).The experimental events were controlled by a computer with 50 times of the maximum injection per day.The times ofactive and inactive nose-poke response and times of drug iniection were recorded per day.Results Compared with group C and P1,the times of active nosepoke response and injections were significantly increased in group P2 and P3(P<0.01).The times of active nosepoke response and injections per day were significantly increased in group P3 than in group P2(P(0.01).There was no significant difference in the times of active nose-poke response and injections between group C and P1.There was no significant difference in inactive nose-poke resporme between the 4 groups.And the total daily doses of propofol injected in the last 3 days were significantly increased in a dose-dependent manner.Conclusion Propefol can induce the development of psychological dependence in rata and it is related to the dosage.
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Objective To investigate the effects of lipoxin A4 on store-operated calcium channel (SOC) and production of reactive oxygen species in macrophages induced by hpopolysaccharide (LPS).Method Macrophages were randomly assigned Io one of the following six groups:control group,LPS group,Thapsigargin group,lipoxin A4+LPS group,lipoxin A4+Thapsigargin group,2-Aminoethoxydiphenylborate+Thapsigargin group.The intracellular[Ca2+]iwas analyzed by eonfoeal laser microscopy.The production of reactive oxygen specips(ROS) was assayed by flow cytometry.Results LPS increased intracellular[Ca2+]i and reactive oxygen species in a dose-dependent manner.Lipoxin A4 suppressed approximately 75% of the Ca2+ ertry signal induced by thapsigargin and suppressed approximately 93% of the Ca2+ entry signal induced by LPS.The increase in intracellular[Ca2+]i was associated with increased ROS production which was abolished in the presence of lipoxin A4.Conclusions These findings indicate that the LPS-indueed intracellular[Ca2*]i increase depends on the Ca2+entry through SOC channel,and lipoxin A4 inhibits Ca2+ influx and ROS production through SOC channel in ratine maerophages induced by LPS.
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To investigate the protective effect of curcumin on endotoxin-induced acute lung injury in rats, and explore the underlying mechanisms, 24 male Wistar rats were randomly divided into 4 experimental groups: sham-vehicle (S), sham-curcumin (C), lipopolysaccharide (LPS)-vehicle (L), and curcumin-lipopolysaccharide (C-L) groups. The wet/dry (W/D) weight ratio of the lung and bronchoalveolar lavage (BAL) fluid protein content were used as measures of lung injury. Neutrophil recruitment and activation were evaluated by BAL fluid cellularity and myeloperoxidase (MPO) activity in cell-free BAL and lung tissue. The levels of cytokine-induced neutrophil chemoattractant-1(CINC-1) in lung tissues were measured by ELISA. The histopathological changes of lung tissues were observed by using the HE staining. Our results showed that lung injury parameters, including the wet/dry weight ratio and protein content in BALF, were significantly higher in the L group than in the S group (P＜0.01). In the L group, higher numbers of neutrophils and greater MPO activity in cell-free BAL and lung homogenates were observed when compared with the S group (P＜0.01).There was a marked increase in CINC-1 levels in lung tissues in response to LPS challenge (P＜0.01,L group vs S group). Curcumin pretreatment significantly attenuated LPS-induced changes in these indices. LPS caused extensive morphological lung damage, which was also lessened after curcumin pretreatment. All the above-mentioned parameters in the C group were not significantly different from those of the S group. It is concluded that curcumin pretreatment attenuates LPS-induced lung injury in rats. This beneficial effect of curcumin may involves, in part, inhibition of neutrophilic recruitment and activity, possibly through inhibition of lung CINC-1 expression.
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To investigate the protective effect of curcumin on endotoxin-induced acute lung injury in rats, and explore the underlying mechanisms, 24 male Wistar rats were randomly divided into 4 experimental groups: sham-vehicle (S), sham-curcumin (C), lipopolysaccharide (LPS)-vehicle (L), and curcumin-lipopolysaccharide (C-L) groups. The wet/dry (W/D) weight ratio of the lung and bronchoalveolar lavage (BAL) fluid protein content were used as measures of lung injury. Neutrophil recruitment and activation were evaluated by BAL fluid cellularity and myeloperoxidase (MPO) activity in cell-free BAL and lung tissue. The levels of cytokine-induced neutrophil chemoattractant-I (CINC-1) in lung tissues were measured by ELISA. The histopathological changes of lung tissues were observed by using the HE staining. Our results showed that lung injury parameters, including the wet/dry weight ratio and protein content in BALF, were significantly higher in the L group than in the S group (P<0.01). In the L group, higher numbers of neutrophils and greater MPO activity in cell-free BAL and lung homogenates were observed when compared with the S group (P<0.01). There was a marked increase in CINC-1 levels in lung tissues in response to LPS challenge (P<0.01, L group vs S group). Curcumin pretreatment significantly attenuated LPS-induced changes in these indices. LPS caused extensive morphological lung damage, which was also lessened after curcumin pretreatment. All the above-mentioned parameters in the C group were not significantly different from those of the S group. It is concluded that curcumin pretreatment attenuates LPS-induced lung injury in rats. This beneficial effect of curcumin may involves, in part, inhibition of neutrophilic recruitment and activity, possibly through inhibition of lung CINC-1 expression.
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OBJECTIVE： A RP－ HPLC method was established for determination of serum concentration of propofol in man METHODS： The mobile phase consisted of methanol－ water（ 68∶ 32v/v） The detection was carried out at wave length 258nm and flow rate 1ml/min with carbamazepine as internal standard RESULTS： The retention times of propofol and carbamazepine were 9 20 and 5 16 min， respectively The mean recovery of propofol was 99％ The within－ day and inter－ day variations were all less than 10％ Propofol and carbamazepine were seperated well The assay linearity was obtained in the range of 1～ 16μ g/ml in serum（ r=0 9 994） CONCLUSION： The method is sensitive， simple and reliable for the determination of propofol concentration
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OBJECTIVE:A RP-HPLC method was established for determination of serum concentration of propofol in man METHODS:The mobile phase consisted of methanol-water(68∶32v/v)The detection was carried out at wave length 258nm and flow rate 1ml/min with carbamazepine as internal standard RESULTS:The retention times of propofol and carbamazepine were 9 20 and 5 16 min,respectively The mean recovery of propofol was 99% The within-day and inter-day variations were all less than 10% Propofol and carbamazepine were seperated well The assay linearity was obtained in the range of 1～16?g/ml in serum(r=0 9 994) CONCLUSION:The method is sensitive,simple and reliable for the determination of propofol concentration
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AIM: To investigate the effect of epinephrine on LPS-induced pro-inflammatory mediators (TNF-?, NO and COX-2) and anti-inflammatory mediators (HO-1 and IL-10) production in murine macrophage RAW264.7 cells, and to determine whether these effect is due to the influence of epinephrine on NF-?B activation. METHODS: RAW264.7 cells were cultured in vitro with 10 ?g/L LPS in the absence or presence of epinephrine at variant concentrations (1, 5, 10, 50 ?mol/L) for 24 hours, then the supernatants was collected for measuring TNF-? and IL-10 by ELISA and Griess reagent was used to measure NO (NO_2-/NO_3-) concentration. At the same time point, cells were harvested and COX-2, HO-1 and I?B-? was detected by Western blotting. RESULTS: 10 ?g/L LPS significantly induced the production of TNF-?, NO (NO_2-/NO_3-), COX-2, HO-1 and IL-10. When epinephrine was added into the medium together with LPS, the pro-inflammatory mediators production was decreased in a dose-dependent manner, however, anti-inflammatory mediators HO-1 and IL-10 expression was enhanced by epinephrine. Epinephrine has no significant effect on I?B-? degradation in LPS-activated RAW264.7 cells. CONCLUSION: Epinephrine down-regulates LPS-induced pro-inflammatory mediator expression while promotes anti-inflammatory mediator production in murine macrophages. These effect seems to be independent of NF-?B activation.