Objective To explore the effect of panaxadiols saponin (PDS) on the oxidative damage in lung tissue in hemorrhage-lipopolysaccharides (LPS) two hits rats. Methods Forty Wistar rats were divided randomly into 4 groups: sham operational group(S group); two hits groups with hemorrhage-LPS(HL group);dexamethasone preventive therapy group (HLD group) ;panaxadiol saponins preventive therapy group(HLP group). The rats were made first-hit by hemorrhagic shock,LPS were administered intraperitoneally to make endotoxic shock and induce two hits in rats,then the model of acute lung injury (ALI) induced by two hits had been built up successfully. NO-2/NO-3,MDA content and NOS,iNOS,SOD activities were measured by 722S spectrophotometer. Results The results of pathological observation showed that there was obvious inflammatory reaction in lung tissues after two-hits,and the structure was destroied.Compared with HL group,the inflammatory reaction was reduced in HLD group and HLD group.NOS,iNOS activities and NO-2/NO-3,MDA contents were increased significantly,and SOD activity was significantly lower in HL group compared with S group(P

AIM: To establish the two-hit rat model with hemorrhage and lipopolysaccharide(LPS) and to study the effect of panaxadiol saponins(PDS) against acute lung injury.METHODS: Forty Wistar rats were divided randomly into 4 groups: sham operational group(S);two-hit groups with hemorrhagic shock-LPS(HL);dexamethasone pretreatment group(HLD) and PDS pretreatment group(HLP).The mean arterial blood pressure(MABP) was monitored dynamically by 4-channel physiological meter RM-6000,and pathological alteration of lung tissues was also observed.The levels of various serum enzymes,TNF-? and IL-6 were detected.RESULTS: MABP decreased in two-hit rat model with hemorrhage-LPS.The serum levels of aspartate aminotransferase,alanine aminotransferase,alkaline phosphatase,lactate dehydrogenase,creatine kinase,TNF-? and IL-6 increased significantly.Severe inflammatory change of pulmonary interstitium in HL group was also observed.CONCLUSION: Endotoxin injection following hemorrhage can be used to establish the animal model of acute lung injury.Similar with dexamethasone,PDS prevents lung tissue from seriously damage through increasing MABP and decreasing the level of serum enzymes,TNF-? and IL-6 levels.

Objective To investigate the alteration of aquaporin 1(AQP1) expression in lung tissues in hemorrhage-lipopolysaccharide(LPS) two-hits rats and the effects of panaxadiols(PDS)and dexamthasone(Dex) on it.Methods The rat model of acute lung injury was built with hemorrhage-LPS two hits.The experiment was divided into control group(S),two-hits model group(HL),DEX group(HLD),and PDS group(HLP).The pathological changes of lung tissue were examined by HE staining.The expression of AQP1 was analyzed by RT-PCR,Western blotting and immunohistochemical staining.Results ① Significant inflammatory changes in pulmonary interstitial of rats in HL group were observed.However,in HLD group and HLP group,the pulmonary pathologic changes were much slighter.② AQP1 mRNA and protein expressions in lung tissues in HL group were significantly decreased compared with others groups(P

The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24 +/- 0.11 and 0. 42 +/- 0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P < 0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9 +/- 9.9 vs 44.8 +/- 15.3, P < 0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P < 0.01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.
Bronchi/metabolism ; Bronchi/pathology ; Cells, Cultured ; HSP70 Heat-Shock Proteins/*biosynthesis ; HSP70 Heat-Shock Proteins/genetics ; Muscle, Smooth/cytology ; Pulmonary Disease, Chronic Obstructive/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Smoking

0.05),but the contraction of the bronchial rings caused by histamine,potassium chloride,intracellular Ca2+ release and extracellular Ca2+ influx was signifi cantly inhibited by Shenmai injection(compare with those of control,P

AIM:To investigate the effect of human Kv1.5 gene transfection on the proliferation and apoptosis in human airway smooth muscle cells(HASMCs).METHODS:Kv1.5 gene was transiently transfected into HASMCs with Lipofectamine 2000 and the level of Kv1.5 protein was observed by Western blotting.Ca 2+ concentration in HASMCs was investigated by fluorescent quantitation using fluorospectrophotometer.Flow-cytometry,MTT method and TUNEL were used to detect the effects of Kv1.5 gene transfection on proliferation and apoptosis in HASMCs.RESULTS:(1)Western blotting showed that transfection of human Kv1.5 plasmid significantly increased the Kv1.5 protein expression compared to control group(P

AIM: To investigate the role of intracellular free Ca2+ concentration ([Ca2+]i) in the regulation of calcium-activated chloride (ClCa) channels in pulmonary artery smooth muscle cells (PASMCs) of rats under normoxic, acute and chronic hypoxic conditions. METHODS: Acute hypoxia-induced contraction was observed in rat pulmonary artery by using routine blood vascular perfusion in vitro. The fluorescence Ca2+ indicator Fura-2/AM was used to observe [Ca2+]i of rat PASMCs in normal and chronic hypoxic condition. The influences of ClCa channels on PASMCs proliferation were assessed by MTT assay. RESULTS: (1) The ClCa channel blockers niflumic acid (NFA) and indaryloxyacetic acid (IAA-94) produced inhibitory effects on acute hypoxia-evoked contractions in pulmonary artery. (2) Under chronic hypoxic condition, [Ca2+]i was increased. In normoxic condition, [Ca2+]i was (123.63?18.98) nmol/L, and in hypoxic condition, [Ca2+]i was (281.75?16.48)nmol/L (P0.05). (4) Chronic hypoxic increased [Ca2+]i which opened ClCa channels. The NFA and IAA-94 blocked them and decreased [Ca2+]i from (281.75?16.48)nmol/L to (117.66?15.36)nmol/L (P

AIM: To investigate the contribution of diazoxide,an opener of mitochondrial ATP-sensitive K+ channel(MitoKATP),and mitochondrial membrane potential(??m) to change of H2O2 in rat pulmonary artery smooth muscle cells(PASMCs) and to unbalance between cell proliferation and apoptosis of PASMCs induced by hypoxia.METHODS: The rat PASMCs were isolated from fresh normal lung tissues and cultured,which were divided into 6 groups,as follows: ① control group;② diazoxide group;③ 5-HD group;④chronic hypoxia group;⑤ chronic hypoxia+diazoxide group;⑥ chronic hypoxia +5-HD group.The relative change in mitochondrial potential was detected with rhodamine fluorescence(R-123) technique.The level of H2O2 in rat PASMCs was detected with chemiluminescence method.The proliferation of rat PASMCs was examined by cell cycle analysis and MTT colorimetric assay.RESULTS: After exposed to diazoxide for 24 h,the intensity of R-123 fluorescence,the level of H2O2 and the A value in normoxic rat PASMCs were significantly increased,and the apoptosis of rat PASMCs was significantly decreased as compared with control group(P

To investigate the relationship between intracellular free Ca2+ concentration ([Ca2+]i) and calcium-activated chloride (Clca) channels of pulmonary artery smooth muscle cells (PASMCs) in rats under acute and chronic hypoxic conditions, acute hypoxia-induced contraction was observed in rat pulmonary artery by using routine blood vascular perfusion in vitro. The fluorescence Ca2+ indicator Fura-2/AM was used to observe [Ca2+]i of rat PASMCs under normal and chronic hypoxic condition. The effect of Clca channels on PASMCs proliferation was assessed by MTT assay. The Clca channel blockers niflumic acid (NFA) and indaryloxyacetic acid (IAA-94) exerted inhibitory effects on acute hypoxia-evoked contractions in the pulmonary artery. Under chronic hypoxic condition, [Ca2+]i was increased. Under normoxic condition, [Ca2+]i was (123.63 +/- 18.98) nmol/L, and in hypoxic condition, [Ca2+]i was (281.75 +/- 16.48) nmol/L (P<0.01). Under normoxic condition, [Ca2+]i showed no significant change and no effect on Clca channels was observed (P> 0.05). Chronic hypoxia increased [Ca2+]i which opened Clca channels. The NFA and IAA-94 blocked the channels and decreased [Ca2+]i from (281.75 +/- 16.48) nmol/L to (117.66 +/- 15.36) nmol/L (P<0.01). MTT assay showed that under chronic hypoxic condition NFA and IAA-94 decreased the value of absorbency (A value) from 0.459 +/- 0.058 to 0.224 +/- 0.025 (P<0.01). Hypoxia increased [Ca2+]i which opened Clca channels and had a positive-feedback in [Ca2+]i. This may play an important role in hypoxic pulmonary hypertension. Under chronic hypoxic condition, Clca channel may play a part in the regulation of proliferation of PASMCs.

Objective To explore the influence of Helicobacter pylori (Hp) on the proliferation, apoptosis and expression of p27kipl protein of gastric epithelial cells in vitro. Methods SC, C-7901 cell pro-liferation was examined by flow cytometry after incubation with different concentration of NCTC11637. The effect of NCTC11637 on cell apoptosis was also evaluated by flow cytometryo And the expression of p27kipl of gastric epithelial cells was detected by immunohistochemical analysis and in situ hybridization, respectively. Results Cell proliferation was enhanced when co-incubated with the Hp in low concentration (3.4 x 104 to 1.9 x 105 CFU/ml) but inhibited in higher concentration (≥4.8×106CFU/ml). However, cell apoptosis was increase when co-incubated with the Hp in high concentration (≥9.6 ×105 CFU/ml) showing concen-tration dependent picture. In addition, the co-incubation of SGC-7901 with Hp led to decrease of the expres-sion of p27kipl protein but not mRNA in a Hp concentration dependent way. Conclusion Hp could effect the gastric epithelial cells apoptosis and proliferation directly and influence the expression of p27kips protein which might facilitate gastric carcinoma.