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ObjectiveTo investigate the mechanism of anti-inflammatory effect of diallyl sulfide (DAS) in protection against acute lung injury (ALI) in rats with paraquat poisoning.Methods Eighty male Wistar rats were randomly divided into four groups, namely: control group, model group, dexamethasone (DXM) treatment group, and DAS treatment group, with 20 rats in each group. The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by gavage, while the same volume of normal saline (NS) was given in same manner in control group. 100 mg/kg of DAS, the same volume of NS, or 1 mg/kg DXM injection were given respectively in DAS treatment group, model group, or DXM treatment group intraperitoneally after exposure to paraquat, once a day for 14 days. Five rats in each group were sacrificed at 1, 3, 7, 14 days, respectively. The inferior lobe of right lung was harvested, and the degree of lung injury was observed with hematoxylin and eosin (HE) staining under optical microscope; the upper lobe of right lung was used to determine the lung wet/dry weight (W/D) ratio and for evaluation of the degree of pulmonary edema. The expression of nuclear factor -κB (NF-κB) in the middle lobe of right lung was assessed with immunohistochemistry. The expression of tumor necrosis factor -α (TNF-α) mRNA in the left lung was determined with the reverse transcription-polymerase chain reaction (RT-PCR).Results① The pulmonary structure in control group was found to be intact. However, in the model group there were progressive pathological changes in lung, including marked edema and thickening of alveolar walls, collapse of alveoli, infiltration of inflammatory cells, alveolar wall, and obvious bleeding in the local lung tissue, and formation of transparent membrane in alveolar space. Less infiltration of inflammatory cells and no obvious destruction were found in alveolar structure in the DAS and DXM treatment groups.② Lung W/D ratio: lung W/D ratio of model group was apparently higher than that in control group at every time point, and peaking on the 3rd day (6.15±0.54 vs. 4.15±2.10,P< 0.05), and the ratio of lung W/D of DAS and DXM treatment groups was obviously lower than that in model group at every time point, especially on the 3rd day (3.99±1.26, 4.30±0.70 vs. 6.15±0.54, bothP< 0.05), but there was no significant difference between DAS and DXM treatment groups in this regard.③ The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell nuclei of the control group, while extensive NF-κBp65 expression was found in model group. Minimal NF-κBp65 positive expression in the cytoplasm and even less positive expression in the nucleus was found in the DAS and DXM treatment groups, and integralA value was significantly lower in the DAS and DXM treatment groups than that of the model group, especially on the 3rd day [(17.98±0.06)×107, (18.53±0.04)×107 vs. (28.85±0.61)×107, bothP< 0.01], but there was no significant difference between DAS and DXM treatment groups.④ It was shown by RT-PCR that the expression of TNF-α mRNA in lung tissue of the model group was significantly higher than that in the control group on the 3rd day (gray value: 3.63±0.62 vs. 0.51±0.13, P< 0.05). The expression of TNF-α mRNA in lung tissue was significantly decreased in DAS and DXM treatment groups compared with model group (gray value: 2.49±0.57, 2.02±0.26 vs. 3.63±0.62, bothP< 0.05), but there was no significant difference between DAS and DXM treated groups.ConclusionTreatment with an intraperitoneally injection of DAS is capable of attenuate the extent of PQ-induced ALI in rats by alleviating pulmonary edema, inhibiting the expression of NF-κB and TNF-α in lung tissue, and ameliorating pathological changes in lung tissue.
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Objective To explore the effectiveness of ω-3 fatty acids in intervening rat models of chronic obstructive pulmonery disease (COPD).Methods The rat COPD models were established by cigarette smoking and intratracheal lipopolysaccharide instillation.Totally 90 rats were randomly divided into 3 groups:normal control group (treated with normal saline),COPD group,and intervention group (the COPD rat models treated with ω-3 fatty acids).Lung tissues were obtained on the 7th,21st,and 28th day.The left lower lobes were analyzed to determine the expressions of nuclear factor-kappa beta (NF-κB) and interferon-γ (IFN-γ)and the right lung lobes were sliced for detecting the cell apoptosis.Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was used to detect the serum IFN-γ and interleukin-6 (IL-6).Results (1) The pathological changes of lung tissue:there were a large number of inflammatory exudation,alveolar wall thickening,hyperplasia of vascular smooth muscle and the alveolar structure destruction in the COPD model group,but the lung tissue were part of alveolar cavity and a little inflammatory exudate in ω-3 fatty fish acids treatment group,control rats were almost no alveolar inflammation on the 28th days.(2) On the 28th day,NF-κB protein expression of the lung tissue (18.91 ± 3.07) in rats of COPD model group was significantly higher than the control group and the intervention group (5.47 ±4.86 and 7.23 ±2.21) (P ＜0.01).On the 28th day,IFN-γ protein expression in lung tissue of the rats in the model group was 7.12 ±3.37,significantly lower than the intervention group (18.74 ± 2.65),the difference was statistically significant (P ＜ 0.01).(3) The IL-6 levels of the blood-serum of model group rats were (13.43 ± 2.47) ng/L,significantly higher than the control group and the intervention group [(4.78 ± 1.93) and (4.98 ± 1.89) ng/L],the differences were statistically significant (P ＜ 0.01) on the 28th day,,but the IFN-γ level [(2.23 ± 0.63) ng/L] in COPD group was more poorer than ω-3 fatty acids group and the intervention group [(4.51 ± 0.71) and (7.05 ± 0.52) ng/L] (P ＜ 0.01).Conclusions The ω-3 fatty acids can lower NF-κB protein expressions in lung tissues and serum and IL-6 levels in COPD rats; aslo,it can increase the IFN-γ protein expression in lung tissue and serum.Thus,it can prevent the lung inflammation in COPD rats.
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ObjectiveTo investigate the mechanism of protective effect of Xuebijing injection on vascular endothelial cells in rats with heat stress.Methods Ninety Sprague-Dawley(SD) rats were randomly divided into control, model and Xuebijing injection treatment groups, 30 rats in each group. Heat stress model was reproduced by placing rats in constant temperature box at 40℃, 60% relative humidity for 1 hour, Xuebijing injection group was treated by intraperitoneal injection of Xuebijing 2.5 g/kg, while the control and model groups were treated by intraperitoneal injection of normal saline 2 mL/kg, once a day only in 1 day for both groups. After model establishment, the rectum temperature, heart rate and the mean arterial pressure(MAP) were recorded at 2, 6, 12 hours in each group. At the same time, the rat abdominal aortic blood was collected and serum was separated, the enzyme-linked immunosorbent assay(ELISA) was used to determine the aortic serum levels of lipopolysaccharide(LPS), nuclear factor-κB(NF-κB) and p53, and the prothrombin time(PT), activated partial thromboplastin time(APTT) and D-dimer of venous blood were detected by automatic blood coagulation analyzer(ACLTOP).Results Compared with those in control group, the rectum temperature, heart rate, LPS, NF-κB, p53, PT, APTT, D-dimer were significantly increased, and MAP was obviously decreased in model group(P<0.05 orP<0.01). Compared with model group, the above indexes were improved significantly in Xuebijing injection treatment group at 2 hours〔rectum temperature(℃): 38.02±0.22 vs. 39.32±0.33, heart rate(bpm): 507±14 vs. 562±35, MAP(mmHg, 1 mmHg=0.133 kPa): 98±6 vs. 87±13, LPS(ng/L): 0.65±0.03 vs. 0.82±0.05, NF-κB(ng/L): 1.10±0.04 vs. 1.33±0.05, p53(ng/L): 1.33±0.03 vs. 1.73±0.02, PT(s):15.47±1.03 vs. 20.28±2.01, APTT(s): 40.26±2.46 vs. 47.46±3.51, D-dimer(μg/L): 238.54±8.32 vs. 323.12±8.14,P<0.05 orP<0.01〕.Conclusion Xuebijing injection can correct the disorders of blood PT, APTT, D-dimer via decreasing the secretion of the levels of NF-κB, p53 from vascular endothelial cells in rats with heat stress, thus the integrity of the vascular endothelium can be protected, and LPS entering into the blood stream can be inhibited.
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ObjectiveTo compare the effectiveness of ω-3 fatty acids and middle/light fatty acids in the intervention of rats model of bleomycin-induced pulmonary fibrosis.MethodsTotally 120 rats were randomly divided into 4 groups: normal saline (NS) group,bleomycin-induced pulmonary fibrosis without treatment group ( BLM group),middle/light fatty acids group,ω-3 fatty acids group.Lung tissues were obtained on the 7th,14th,and 21st day after modeling.The left lung were measured by using immunohistochemical methods for transforming growth factor β1 (TGF-β1) and interferon garmma ( IFN-γ).The lower lobe of the right lung underwent HE staining.Serum TGF-β1,IFN-γ,and interleukin-4 (IL-4) were measured using double-antibody sandwich ELISA.ResultsThe pulmonary alveolitis and fibrosis in the ω-3 fatty acids group was significantly milder than in middle/light fatty acids group and BLM group.On the 7th,14th,and 21st day after modeling,stronger TGF-β1 protein expression was detected in the bronehiolar epithelia of middle/light fatty acids group and BLM group and poorer IFN-γexpression in both groups.However,the opposite results were found in the ω-3 fatty acids group: on the 7th,14th,and 21st day after modeling,TGF-β1 protein expression ( 13.60 ± 5.90,10.53 ± 4.21,and 7.23 ± 2.21 )was lower ( P =0.047) and IFN-γ ( 13.85 ± 7.48,15.32 ± 2.12,and 18.74 ± 2.65 ) was stronger in ω-3 fatty acids group (P =0.041).On the 7 ,14,and 21st day after modeling,the serum IL-4 levels in the middle/light fatty acids group and BLM group became significantly higher,while the IFN-γ level in both groups was significantly lower than in ω-3 fatty acids group ( P =0.008 ) ; meanwhile,in the ω-3 fatty acids group,the serum IL-4 levels [ (8.73 ± 1.20),(5.73 ±2.03),and (4.98 ± 1.89) pg/ml] were significantly lower (P =0.044) and serum IFN-γlevels [ (5.67 ± 0.13 ),( 6.58 ± 0.64 ),and ( 7.05 ± 0.52 ) pg/ml ] were significantly higher ( P =0.048 ) on the 72,14th,and 21st day after modeling.Conclusionsω-3 fatty acids can lower TGF-β1 protein expression in rat lung tissue and reduce the surum TGF-β1 and IL-4 levels.Compared with the middle/light fatty acids,it can more effectively upregulate the expression of IFN-γ in lung tissue and increase its serum level,and thus alleviate pulmonary fibrosis in rats.