Protective effect of Auricularia auricular-judae polysaccharide on pulmonary tissues of rats with LPS-induced acute lung injury

AIM: To investigate the effects of Auricularia auricular-judae polysaccharide(AAP) on pulmonary tissues of rats with LPS-induced acute lung injury(ALI) and its mechanisms.METHODS: Adult Sprague-Dawley rats were randomly divided into control group, LPS group,low-dose AAP group, middle-dose AAP group, high-dose APP group, and dexamethasone group. The rats were injected with LPS(8 mg/kg, ip) to induce ALI. The rats in the AAP groups were treated with AAP for 7 d before the induction of ALI. The protein concentration in the bronchoalveolar lavage fluid(BALF) was measured. The lung edema degree was measured by detecting the wet/dry weight ratio. The myeloper-oxidase(MPO), total antioxidant capacity(T-AOC), total superoxide dismutase(T-SOD), nitric oxide synthase(NOS) and malondialdehyde(MDA) levels were determined. The pathological changes of the lung tissues were evaluated by HE staining.RESULTS: Treatment with AAP significantly improved LPS-induced lung pathological changes, attenuated the protein concentration in the BALF and wet/dry weight ratio, inhibited the activities of MPO and NOS, reduced MDA level and increased the activities of T-AOC and T-SOD.CONCLUSION: AAP protects against LPS-induced acute lung injury in rats.     

Sodium nitrite reduces lipopolysaccharide-induced acute lung injury in mice

AIM: To investigate the effect of sodium nitrite (SN) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and its underlying mechanism in mice. METHODS: All male Institute of Cancer Research (ICR) mice were randomly divided into five groups: Control group; LPS group; and SN 4.8 nmol/L, SN 48 nmol/L, SN 480 nmol/L (ip) groups. Lung wet weight/dry weight (W/D) ratio and permeability were detected. Neutrophil infiltration in bronchoalveolar lavage fluid (BALF) was measured by cel1 counting and morphological changes in lung tissues were assayed by hematoxylin-eosin staining. The 1evels of interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in lung were detected. Nitric oxide (NO) level and nitric oxide synthase (NOS) activity in lung were measured according to the specification. RESULTS: Compared to lung in LPS-induced ALI mice, at doses of 4.8 nmol/L and 48 nmol/L, not 480 nmol/L, SN markedly decreased the lung W/D ratio, total leukocyte number and neutrophil percentage in the BALF, lung permeability, and TNF-α/IL-10 ratio, in lung. SN at dose of 480 nmol/L markedly increased the lung NO level compared to control group. In addition, SN decreased the total NOS and inducible NOS (iNOS) activities compared to LPS-induced ALI mice. CONCLUSION: These results indicate that the protective effect of SN against LPS-induced ALI in mice is associated with the low dose SN-induced NO, as well as the subsequent decrease in iNOS activity and TNF-α/IL-10 ratio.     

Protective effect of recombinant human serum albumin-thioredoxin fusion protein on mice with acute lung injury induced by influenza virus

AIM To investigate the protective effect of recombinant human serum albumin (HSA)-thioredox?in (Trx) fusion protein (HSA-Trx) on mice with acute lung injury (ALI) induced by influenza virus infection. METH?ODS: The recombinant HAS-Trx fusion protein was generated by Pichia pastoris expression system. ICR mice were used to establish the animal model of ALI induced by PR8 (H1N1) influenza virus, and the experimental mice were divided into healthy control group, ALI group, ALI+Trx group and ALI+HSA-Trx group, with 10 mice in each group. The bronchoalveolar lavage fluid (BALF) in each group was collected, the total number of cells and the number of alveolar neutrophils were determined, the protein concentration was measured by Coomassie brilliant blue solution method, and the interferon-γ (IFN-γ) content in BALF was detected by ELISA. The lung tissues were collected for hematoxylin and eosin staining. The inducible nitric oxide synthase (iNOS), 8-hydroxydeoxyguanosine (8-OHdG) and 3-nitrotyrosine (NO₂-Tyr) in lung tissues were detected by immunofluorescence method. Peroxide concentration in plasma was evaluated using a CR2000RC analyzer. RESULTS HSA-Trx treatment significantly reduced the total number of cells, neutrophils and total protein in BALF of ALI mice (P<0.05), and decreased the levels of 8-OHdG, NO₂-Tyr in lung tissue and peroxide in plasma (P<0.05). However, it has no significant inhibitory effect on iNOS and IFN-γ expression (P>0.05). CONCLUSION HSA-Trx inhibits inflammatory response and excessive production of nitric oxide in the lung, thus protecting influ?enza virus-induced ALI mice.     

Mechanism of exogenous nitric oxide in attenuating endotoxin-induced increase of rat pulmonary microvascular permeability

AIM and METHODS:The animal model of acute lung injury (ALI) caused by intratracheal instillation of lipopolysaccharides(LPS) in vivo and human peripheral blood polymorphonuclear neutrophil (PMN) in vitro were used to study the effects of sodium nitroprusside (SNP), nitric oxide (NO) donor, on LPS-induced PMN accumulation, microvascular permeability and PMN apoptosis. RESULTS:①In vivo, PMN accumulation in lung, the protein content in bronchoalveolar lavage fluid (BALF) and the Evans blue dye and monastral blue dye extravasation in lung tissue of LPS group were markedly higher than those of both sham operation group and LPS+SNP group. ②In vitro, the apoptotic percentage of SNP group was much higher than that of control group, while compared with LPS group, SNP+LPS group has significantly higher apoptotic percentage. CONCLUSIONS:SNP intratracheal instillation attenuated LPS-induced microvascular permeability and alleviated ALI. PMN apoptosis induced by SNP may be one of the potential mechanisms underlying the decrease of PMN accumulation in lung tissue.     

Protective effect of HSF1 on mice with LPS-induced acute lung injury and screening of relevant differentially-expressed genes

AIM: To study the protective effect of heat shock factor1 (HSF1) on the mice with lipopolysaccharide (LPS)-induced acute lung injury (ALI), and to screen the relevant differentially-expressed genes. METHODS: ALI mouse model was established by LPS intracheal instillation. The macroscopic and pathological changes of the lung tissue were observed, and the concentrations of total protein, TNF-α, IL-β, IL-6 and VEGF in the bronchoalveolar lavage fluid (BALF) were analyzed. Differentially-expressed genes in the lung tissues of HSF1^+/+ mice and HSF1^-/- mice with ALI induced by LPS were screened by gene chips. The key gene was verified by real-time qPCR. RESULTS: The macroscopic and pathological changes of the lung injury in HSF1^-/-+LPS mice were more serious than those in HSF1^+/++LPS mice. The concentrations of total protein, VEGF, TNF-α, IL-1β and IL-6 in the BALF of HSF1^-/-+LPS mice were significantly higher than those of HSF1^+/++LPS mice (P<0.05). Compared with the HSF1^+/+ mice, a total of 918 differentially-expressed genes were indentified in the HSF1^-/- mice, among which the expression levels of 65 genes had obvious diffe-rence, with 28 genes up-regulated, including Atg7, ccr1, cxcr2, Tbl1xr1, Mmp9, Pparg, Plcb2, Arrb2, Cntn1, Col4a6, etc, and 37 genes down-regulated, including Fgfr1, Fgfr2, Map4k4, Ddx58, Tfg, Stat3, Smad4, Lamc1, Sdc3, etc. The results of real-time qPCR showed that the mRNA level of CXCR2 in HSF1^-/-+ LPS mice was significantly higher than that in HSF1^+/++ LPS mice, which was consistent with the results of gene chips. CONCLUSION: HSF1 has protective effect on the mice with LPS-induced ALI. CXCR2 may be involved in the protective effect of HSF1 on this process.     

Protective role of endogenous hydrogen sulfide in cholecystokinin octapeptid against acute lung injury induced by lipopolysaccharide

AIM: To explore the role of endogenous hydrogen sulfide (H₂S) in the mechanism of cholecystokinin octapeptide (CCK-8) to alleviate acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: Eighty-four Sprague-Dawley rats were randomly divided into seven groups: control, LPS (instilled intratracheally to reproduce the model of ALI), NaHS (H₂S donor) +LPS, propargylglycine ［inhibitor of cysathionine-γ-lyase (CSE), PPG］+LPS, CCK-8+LPS, PPG+CCK-8+LPS and CCK-8 group. Animals were sacrificed at 4 h and 8 h after agent instillation. The wet and dry ratio (W/D) of the lung weight was measured and calculated. Morphological changes of lung tissues were observed. H₂S concentration in plasma, malondialdehyde (MDA) content, myeloperoxidase (MPO) and CSE activities in the lung were determined. Furthermore, the level of P-selectin of lung tissue was measured by radioimmunoassay, the CSE mRNA expression in the lung was detected by RT-PCR, and the protein content in bronchoalveolar lavage fluid (BALF) was detected. RESULTS: Compared with control, severe injury of lung tissues and increase in W/D, protein content in BALF, MDA content, MPO activity and P-selectin level in the lung were observed in rats treated with LPS. LPS also lead to a drop in plasma H₂S concentration, lung CSE activity and CSE mRNA expression. Administration of NaHS before LPS could attenuate the changes induced by LPS, while H₂S concentration, CSE activity and CSE mRNA expression were higher than those in LPS group. However, pre-treatment with PPG exacerbated the lung injury induced by LPS, H₂S concentration, CSE activity and CSE mRNA expression were lower than those in LPS and CCK-8 +LPS group, respectively. CONCLUSION: CCK-8 attenuates LPS-induced acute lung injury by means of anti-oxidation and inhibition of PMN adhesion and aggregation, both of which are mediated by endogenous H₂S.     

Mechanisms for protection of berberine against LPS-induced acute lung injury in mice

AIM:To investigate the mechanisms by which berberine attenuates LPS-induced acute lung injury, and provide a new strategy for the treatment of the lung injury due to LPS. METHODS:BALB/c mice were randomly assigned into three groups (control, LPS group, and berberine treatment group). Mice were administered intragastrically with distilled water (0.1 mL/10 g) or neutral sulfate berberine (50 mg/kg) once a day for 3 days, 1 h after intragastrical treatment on day 3, LPS (20 mg/kg) or normal saline was injected intraperitoneally (ip). All animals were sacrificed 12 h after LPS injection, the left lung tissue sections were prepared for histology analysis and the right lung were used to determine the ratio of wet to dry lung tissue weight (W/D). In another experiment, bronchoalveolar lavage fluid (BALF) was collected, and then the total protein content, and the amounts of white blood cells (WBC) and polymorphonuclear neutrophils (PMN) in BALF were determined. Furthermore, the phosphorylation of cytosolic phospholipase A₂ (cPLA₂) was detected with immunohistochemical analysis by using phospho-cPLA2(Ser505) antibody, and the contents of thromboxane B₂ (TXB₂) in BALF, malondialdehyde (MDA) in the lungs, and activity of superoxide dismutase (SOD) in lung tissues were also determined.RESULTS:LPS induced acute lung injury, activated cPLA₂, and increased TXB₂ content in the BALF and MDA level in the lung tissue. The pretreatment with berberine significantly attenuated lung injury, lung edema and protein leakage induced by intraperitoneal injection of LPS. The expression of phospho-cPLA₂ in the lung tissues and TXB₂ content in the BALF in the berberine treatment group were lower than those in LPS group (P<0.05). In addition, the content of MDA in the lung tissue was lower in the berberine treatment group than LPS group (P<0.05), but there was no significant difference in activity of lung SOD between the berberine treatment and LPS group (P>0.05). CONCLUSION:Pretreatment with berberine remarkably reduces the LPS-induced lung injury, which is, at least in part, through inhibiting phosphorylation of cPLA₂ and decreasing lipid peroxidation. These findings provide a new strategy for the prevention and treatment of LPS-induced acute lung injury.     

Effect of interleukin-10 on attenuating endotoxin-induced acute lung injury

AIM: To study the role of polymorphonuclear neutrophile(PMN) in lipopolysaccharide (LPS)- induced acute lung injury (ALI) and the protective effect of interleukin -10(IL-10) on ALI. METHODS: LPS alone (100μg) or LPS+ IL-10 (l ug) was instilled intratracheally into rats. PMN numbers, protein content and malondialdehyde (MDA) content in bronchoalveolar lavage fluid (BALF) were measured. Histological change of lung was also observed. RESULTS: LPS increased significantly PMN numbers, protein content and MDA content in BALF. Histological finding shows PMN accumulation in lung. IL - 10+LPS reduced remarkably PMN numbers ,pro- tein content and MDA content in BALF than those caused by LPS. PMN decreasing was also identified by light microscopy. CONCLUSION: LPS instilled intratracheally causes PMN accumulation in lung and ALI, while IL - 10 could alleviate ALI through reducing PMN accumulation.     

Bone marrow-derived mesenchymal stem cell-conditioned medium atte-nuates LPS-induced acute lung injury in mice

AIM:To explore the effects of bone marrow-derived mesenchymal stem cells-conditioned medium (MSCs CdM) on lipopolysaccharide (LPS)-induced acute lung injury. METHODS:Lung injury was induced in mice by intraperitoneal injection of LPS. The mice were given a tail vein injection of MSCs CdM or normal saline 1 h after LPS administration. The mice were killed by an intraperitoneal injection of pentobarbital 6 h after LPS injection for either bronchoalveolar lavage fluid (BALF) and serum collection or lung histological analysis. RESULTS:Compared with control group, the BALF levels of protein, interleukin-10 (IL-10) and keratinocyte growth factor (KGF), the serum levels of tumor necrosis factor α (TNF-α) and IL-6, and the myeloperoxidase (MOP) activity in the lung tissues were significantly higher in LPS group, and severe pathological damages in the lung tissues were also observed. Treatment with MSCs CdM significantly reduced the BALF prtein level, the seum TNF-α and IL-6　levels and the lung MPO activity, and attenuated the lung pathological damages, but further increased the levels of IL-10 and KGF in the BALF. CONCLUSION:Treatment with MSCs CdM attenuates the lung injuries induced by LPS, which may be via regulating the expression of TNF-α, IL-6, IL-10 and KGF.     

Role of hydrogen sulfide in acute lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To explore the role of endogenous and exogenous hydrogen sulfide (H₂S) in acute lung injury (ALI) induced by ischmia-reperfusion (IR) of hind limbs in rats.METHODS: A Sprague-Dawley rat model of acute lung injury was induced by ischemia of the hind limbs for 4 h and reperfusion for another 4 h. The rats (n=120) were randomly divided into 4 groups: control, IR, NaHS (H₂S donor)+IR, and propargylglycine +IR. The animals were sacrificed after reperfusion. Lung weight/body weight ratio (LW/BW) was measured and calculated. Morphological changes of the lung tissues were observed. The concentrations of H₂S, nitric oxide (NO) and carbon monoxide (CO) in plasma were tested. The content of malondialdehyde (MDA), the activity of CSE, inducible nitric oxide synthase (iNOS) and hemeoxygenase (HO) in the lungs were determined. The polymorpho-nuclear neutrophils(PMN) and protein content in bronchoalveolar lavage fluid(BALF) were also measured. The correlation of H₂S content with the above indices was analyzed.RESULTS: Compared with control group, severe injuries of the lung tissues, raised LW/BW, MDA concentration, PMN and protein contents in BALF were observed in IR group. Limb IR also made a drop in the concentration of plasma H₂S and the activity of lung CSE, while the activity of iNOS and HO in the lung tissues and the levels of plasma NO and CO increased. Administration of NaHS before IR attenuated the changes induced by IR, while pre-administration of PPG exacerbated the IR injuries and increased the plasma NO level and lung iNOS activity. The H₂S content was positively correlated with CSE activity, CO content and HO-1 activity (P<0.01), and negatively correlated with the other indices (P<0.01).CONCLUSION: Down-regulation of H₂S/CSE is involved in the pathogenesis of acute lung injury induced by IR. Endogenous and exogenous H₂S protects against lung injuries. The anti-injury effects of H₂S are related with its anti-oxidative activity to attenuate the inflammatory over-reactions in the lung induced by PMN. Down-regulation of NO/iNOS system and up-regulation of CO/HO-1 system by H₂S are also involved in the process of anti-injury to ALI.     

Meconium induces formation of nitrotyrosine and expression of inducible nitric oxide synthase in the rat lung

AIM: To evaluate the contribution of inducible nitric oxide synthase (iNOS) and nitrotyrosine to acute lung injury (ALI) in rats with meconium aspiration. METHODS: 16 health male Sprage-Dawley rats were randomized to control group and meconium group, followed by intratracheally administration of 1 mL/kg saline or 1 mL/kg 20% human newborn meconium suspension. The animals were killed after 24 h of treatment. The measurements included bronchoalveolar lavage fluid (BALF) cell count, pulmonary myoloperoxidase (MPO) activity and nitric oxide (NO) level. Western bloting was used to determine the expression of pulmonary nitrotyrosine-a specific “footprint” of peroxynitrite and iNOS. RESULTS: Compared to control group, the rats in the meconium group had increased BALF cell counts ［(4.04±1.01)×109cells/L vs （0.53±0.19)×109cells/L］, pulmonary MPO activity ［(1.49±0.22）U/g wet lung tissue vs (0.62±0.16) U/g wet lung tissue］, NO level ［(12.77±5.00） mmol/g protein vs （4.89±1.32) mmol/g protein］, increased expression of nitrotyrosine and iNOS (0.46±0.19 and 1.49±0.60 vs 0.15±0.04 and 0.09±0.04, respectively), all P<0.01. CONCLUSIONS: Meconium results in an increase in expression of pulmonary iNOS, leading to over production of NO and nitrotyrosine, which may be of pathogenic importance in the ALI with meconium aspiration.     

Sphingosine-1-phosphate receptor 2 attenuates influenza A virus-induced viral pneumonia through PI3K/AKT/eNOS pathway

AIM: To investigate the effects of sphingosine-1-phosphate receptor 2 (S1PR2) on influenza A virus-induced viral pneumonia.METHODS: The animal model of influenza A virus pneumonia was established by infecting wild-type C57BL/6 mice and S1pr2^-/- mice with influenza virus subtype FM1 mouse lung adaptable strain through nose drops. The pathological changes of the lung tissues of wild-type mice (model group), JTE-013 (S1PR2 effective antagonist)-challenged mice and S1pr2^-/- mice were observed, and the protein concentration, total cell number, and interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) levels were determined in the bronchoalveolar lavage fluid (BALF) at 4 d and 6 d after virus infection. The phosphorylation levels of AKT and eNOS in the lung tissues were determined by Western blot. RESULTS: Compared with the wild-type mice of control group, the influenza A virus pneumonia in JTE treatment group and S1pr2^-/- mice were more serious, and the protein concentration, total cell number and inflammatory cytokines in the BALF were remarkably increased. Moreover, the phosphorylation levels of AKT and eNOS, the downstream targets of PI3K, were significantly increased (P<0.01). CONCLUSION: S1PR2 mediates PI3K/AKT/eNOS signaling transduction pathway to regulate NO generation, and inhibit vascular permeability and inflammatory cytokine release, thus attenuating the viral pneumonia induced by influenza A virus.     

Role of heme oxygenase in cholecystokinin octopeptide ameliorating acute lung injury induced by lipopolysaccharide

AIM: To study the role of heme oxygenase (HO)-1 in the mechanism of cholecystokinin-octapeptide (CCK-8) for attenuation of acute lung injury (ALI) induced by lipopolysaccharide (LPS).METHODS: Adult male rats were randomly divided into five groups: control group,LPS group,CCK-8+LPS group,LPS+ Hm (hemin,HO-1 donor) group and LPS+ZnPP (zinc protoporphyrin,specific inhibitor of HO-1) group.PMN number in bronchoalveolar lavage fluid (BALF),the structure of the lung,MDA content,HO-1 activity,the expressions of HO-1 mRNA and protein in the lung were detected respectively.RESULTS: The lung injury in LPS group was observed,at the same time the numbers of PMN,the content of MDA,the activity and the expression of HO-1 were all higher than those in control group (all P<0.05).The degree of lung injury,PMN numbers and MDA content were lower,while the activity and the expression of HO-1 in CCK-8+LPS and LPS+Hm group were higher than those in LPS group (all P<0.05).However,the degree of lung injury,PMN numbers and MDA content were higher,the activity and the expression of HO-1 were lower in LPS+ZnPP than those in LPS group respectively (all P<0.05).CONCLUSION: CCK-8 attenuates the LPS-induced ALI by means of anti-oxidation and inhibits PMN aggregation,which are both mediated by HO-1 partly.     

Adipolin/CTRP12 protects against LPS-induced ARDS by up-regulating alveolar epithelial sodium channel in mice

AIM: To investigate the effect of adipolin/CTRP12 in LPS-induced acute respiratory distress syndrome(ARDS) and its potential regulation on alveolar epithelial sodium channel(ENaC) in mice. METHODS: C57BL/6J mice(n=40) were randomly divided into control group, LPS group, adipolin group and wortmannin(PI3K inhibitor) group with 10 mice in each group using random number table. The pathological changes of the lung tissues were evaluated by HE staining. The alveolar fluid clearance(AFC) was measured by Evans blue-marked albumin, and the concentrations of total protein in bronchoalveolar lavage fluid(BALF) were assessed by bicinchoninic acid(BCA) method. In BALF, the levels of IL-1β and TNF-α were determined by ELISA, and the activity of myeloperoxidase(MPO) was detected by an MPO assay kit. The total cell counts and polymorphonuclear neutrophil(PMN) counts in the BALF were analyzed by Giemsa staining. The mRNA levels of α-ENaC were assessed by qPCR, while the protein levels of α-ENaC and p-Akt were determined by Western blot. RESULTS: Compared with control group, the classic ARDS pathological changes were observed in the mice in LPS group, manifesting by severe pathological lung injury(P<0.05), increases in W/D weight ratio, total protein levels, cell counts, MPO activitiy, and IL-1β and TNF-α levels in the BALF, and decrease in AFC(P<0.05), accompanied by down-regulated levels of α-ENaC and p-Akt in the lung tissues(P<0.05). The deteriorating effects triggered by LPS were significantly reversed by administration of adipolin. However, PI3K inhibitor wortmannin canceled the beneficial effects of adipolin on LPS-induced ARDS, as evidenced by aggravated lung injury, increased levels of W/D weight ratio, protein levels, cell counts, MPO activity, and IL-1β and TNF-α levels in the BALF(P<0.05), and decreased levels of AFC, α-ENaC and p-Akt in the lung tissues. CONCLUSION: Adipolin protects against LPS-induced ARDS in the mice by up-regulating α-ENaC and enhancing AFC via PI3K/Akt signal pathway.     

Panaxadiol saponins and dexamethasone have similar actions on LPS-induced AKI in mice

AIM：To investigate whether the panaxadiol saponins (PDS) and dexamethasone (DEX) have similar effects on lipopolysaccharide (LPS)-induced acute kidney injury (AKI). METHODS：C57BL/6 mice were randomly divided into 4 groups: the control mice received intraperitoneal injection of normal saline; in LPS group, the mice were subjected to intraperitoneal injection of LPS (10 mg/kg); in PDS + LPS group and DEX + LPS group, the mice were injected intraperitoneally with PDS (25.0 mg/kg) and DEX(2.5 mg/kg) 1 h before LPS injection, respectively. The blood was collected from the hearts, and the kidneys were collected for the biochemical and Western blotting analysis 12 h after LPS injection. RESULTS：LPS induced AKI, evidenced by markedly increased blood urea nitrogen (BUN) and creatinine (CREA) contents compared with control group (P<0.01). However, serum contents of CREA and BUN obviously reduced in PDS + LPS group and DEX + LPS groups compared with LPS group (P<0.05). Both PDS and DEX decreased the production of TNF-α and IL-6 by inhibiting renal NF-κB signaling activation. PDS and DEX also down-regulated the expression of inducible nitric oxide synthase, up-regulated the expression of manganese superoxide dismutase and reduced oxidative stress in the kidneys of LPS-challenged mice. In addition, treatment with PDS and DEX significantly increased the nuclear glucocorticoid receptor in the kidneys of LPS-treated mice. CONCLUSION：PDS and DEX have inhibitory effects on LPS-induced AKI mice. However, it is unclear whether PDS reduces LPS-induced AKI via direct action on glucocorticoid receptor.     

Cytokine levels and the discrepancy of dexamethasone intervening in ALI rats induced by hydrochloric acid and lipopolysaccharide

AIM: To explore the cytokines level and the discrepancy of reaction to dexamethasone (Dex) in ALI rats induced by hydrochloric acid (HCl) and lipopolysaccharide (LPS). METHODS: Ninety-six SD rats were divided into six groups at random (n=16 in each group): NS group, HCl group, LPS group, NS+Dex group, HCl+Dex group and LPS+Dex group. Every group was divided into two subgroups: the bronchoalveolar lavage (BAL) subgroup and no bronchoalveolar lavage (NBAL) subgroup. The total leukocytes, PMN%, macrophage%, lymphocyte%, total protein in BALF and the wet/dry of the lung weight were measured. The concentrations of TNF-α, IL-1β, IL-4 and IL-10 in serum and BALF in every group were compared. RESULTS: (1) In the groups of LPS and HCl, the total leukocytes, PMN numbers, the protein concentration in the BALF and W/D were higher than those in control group (P<0.05). Compared to LPS groups, the percentage of macrophage increased in LPS+Dex group (P<0.05). (2) In serum and BALF of both LPS group and HCl group, the concentrations of TNF-α, IL-4 and IL-10 were higher than those in the control (P<0.01). The content of IL-1β in serum of all the groups was undetected. Compared to LPS groups, the concentrations of TNF-α and that of IL-1β decreased in LPS+Dex group (P<0.05). The concentration of IL-10 in LPS+Dex group was higher than that in LPS group (P<0.01). CONCLUSION: The permeability and the inflammatory cytokines in these two models were not consistent. Glucocorticoids play an effective role for resisting ALI induced by LPS but not HCl.     

Effect of endogenous H2S on pulmonary hypertension during acute lung injury induced by LPS and its interaction with NO

AIM: To investigate the effect of H2S on pulmonary artery hypertension during acute lung injury induced by LPS and the interaction between the systems of hydrogen sulfide (H2S)/cystathionine-β-lyase (CSE) and nitric oxide (NO)/nitric oxide synthase (NOS) in this process. METHODS: Seventy-two adult male rats were randomly divided into four groups: control group, LPS group, LPS+L-NAME group and LPS+propargylglycine (PPG) group. Mean pulmonary artery pressure (mPAP) of each rat was examined at 2 h, 4 h, 6 h and 8 h after treatment. H2S and NO contents in plasma, NO content, iNOS, cNOS and CSE activity in lung were measured at 4 h or 8 h after treatment, respectively. Expression of iNOS in lung tissue was also detected by immunohistochemistry technique, and the injury of lung was evaluated with morphological changes under microscope. RESULTS: LPS could induce severe lung injury, and mPAP, NO content, iNOS activity and its protein expression in LPS group significantly increased, but cNOS activity, H2S content and CSE activity decreased compared with those of control group. Administration of L-NAME before LPS could attenuate the changes induced by LPS. Pre-administration of PPG, a CSE inhibitor, exacerbated the injury by LPS, but there was no prominent variation in cNOS activity. CONCLUSION: Reduced endogenous H2S could increase pulmonary artery hypertension during acute lung injury induced by LPS. There is a negative effect between H2S/CSE system and NO/NOS system in this process.     

Improvement and evaluation of chronic bronchitis modeling methods in mice

AIM: To explore a more accurate and reliable pathological model of the chronic bronchitis, which has improved from the former single-factor modeling method of the disease.METHODS: The mice in complex group were treated with lipopolysaccharide(LPS) by tracheal injection on the 1st day and nasal drops on the 14th day, and from the 2nd day to 30th day, the animals were given passive smoking and sulfur dioxide(SO₂) inhalation(except on the 14th day). The mice in SO₂ group were exposed to SO₂ 2 min per day, while in smoking group, the mice were exposed to smoke for about 1 h per day(4 cigarettes each time until one pack of cigarettes were burning up). In LPS group, the mice had tracheal injection of LPS on the 1st day and nasal drops of LPS on the 14th day and 30th day. Every modeling process lasted for 30 days. After modeling, the improvement of chronic bronchitis model was evaluated by testing the general conditions of the mice, analyzing leukocyte count in bronchoalveolar lavage fluid(BALF), and observing the morphological changes of the bronchial and lung tissues.RESULTS: After modeling, the mice in every model group experienced symptoms including wet nose, cough, dry and lusterless hair, arched back and curled-up body, showing inactive, and slow down in response. The mice in complex group gained the lowest weight compared to other groups. From each model group, the inflammatory cells infiltrated evidently around the bronchial walls, especially in the bronchial cavity, and the mucilage secretion in the airway increased. The total number of leukocytes in BALF increased significantly in complex group. The inflammatory cell count in the lung tissue indicated that the mice in complex group had significantly higher levels of inflammatory cell infiltration. Besides, the comparison between smoke group and LPS group was statistically significant.CONCLUSION: Smoking, SO₂ inhalation and LPS injection induce bronchial lung disease in mice, and the complex chronic bronchitis mouse model is a better model with the pathological changes of bronchus, lung tissue and BALF, and pathogenesis of chronic bronchitis.     

Vinpocetine injection attenuates lipopolysaccharide-induced acute lung injury in rats

AIM:To observe the inhibitory effects of vinpocetine injection on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in the rats and to explore the underlying mechanisms.METHODS:Male Wistar rats (n=50) were randomly divided into 5 groups:control group,ALI model group,and low,medium and high doses of vinpocetine treatment groups.The rats in control group were injected with 0.9% NaCl at 5 mL/kg through femoral vein.The rats in ALI model group received LPS at 10 mg/kg through femoral vein.After injected with LPS,the rats in vinpocetine treatment groups received vinpocetine at 0.2 mg/kg,0.7 mg/kg or 1.2 mg/kg via intraperitoneal injection.The pathological changes of the lung tissues were observed under microscope with HE staining.The cell apoptosis in the lung tissues was detected by TUNEL staining.Myeloperoxidase (MPO) activity was measured by the method of spectrophotometry.The protein expression of NF-κB,ICAM-1,VCAM-1,Bax and Bcl-2 was determined by Western blot.RESULTS:Compared with ALI group,administration of vinpocetine significantly attenuated the structural injury of the lung and the infiltration of inflammatory cells.Moreover,vinpocetine decreased cell apoptosis and MPO activity in the lung tissues of ALI rats.In addition,the protein expression of NF-κB,ICAM-1,VCAM-1 and Bax was inhibited after vinpocetin treatment,whereas Bcl-2 expression was increased.CONCLUSION:Vinpocetine attenuates LPS-lung injury by reducing MPO activity and regulating NF-κB,ICAM-1,VCAM-1,Bax and Bcl-2 protein expression.     

Role of p38 MAPK-HSP27 signaling pathway in rat acute lung injury induced by lipopolysaccharide

AIM: To observe the role of p38 mitogen-activated protein kinase (p38 MAPK)-heat-shock protein 27 (HSP27) signaling pathway in lipopolysaccharide-induced acute lung injury (ALI) in rats. METHODS: Wistar rats were randomly divided into control group, ALI group and ALI+SB203580 group. After the experimental model was established, the rats were sacrificed. The pathological changes of the lung and the changes of F-actin and G-actin in the endothelial cells were observed. The ratio of wet weight to dry weight (W/D) of the lung tissues was measured. The protein levels in bronchoalveolar lavage fluid (BALF) were detected. The levels of IL-6 and TNF-α in serum and BALF were tested. The concentrations of p-p38 and p-HSP27 in the lung were determined. RESULTS: In ALI group, the protein levels in BALF and W/D ratio of the lung increased significantly at 2 h. The levels of TNF-α and IL-6 in serum and BALF began to increase at 2 h, which had significant difference as compared with control group. Aleolar epithelial swelling, alveolar walls widening, alveolar interstitial and cavity edema, and the exudation of alveolar inflammation cells, red blood cells and protein were observed in ALI group. The protein levels in BALF and W/D ratio of the lung in ALI+SB203580 group were much less than those in ALI group. The exudation of alveolar inflammation cells, red blood cells and protein, and the interstitial and alveolar edema in ALI+SB203580 group alleviated as compared with ALI group. The expression of p-p38 MAPK and p-HSP27 in the lung at 2 h in ALI group was higher than that in control group. F-actin expression in ALI group obviously increased than that in control group at time points of 0 h and 8 h. Compared with ALI group, the expression of p-HSP27 and F-actin in ALI+SB203580 group was reduced. CONCLUSION: Lipopolysaccharide activates p38 MAPK-HSP27 signaling pathway and induces lung injury. Blockage of p38 MAPK-HSP27 signaling pathway may reduce lung injury.