Phenotype and immune activity of dendritic cells under interleukin-18 intervention

AIM: To investigate the phenotype and immune activity of dendritic cells using interleukin-18 as intervent.METHODS: Monocytes were isolated from human peripheral blood and induced into DCs with GM-CSF and IL-4. The cellular morphous was observed under inverted microscope. On the 5th day, 3 groups including IL-18 group, TNF-α group and IL-18+TNF-α group were set. IL-18, TNF-α or IL-18+TNF-α was used as intervents respectively to facilitate cell maturity. Supernatants were collected at 24 h, 48 h and 72 h. IL-12 in the supernatant, CD1a, HLA-DR, CD83 and CD86 were analyzed using flow cytometry. DCs of the 3 groups were co-cultured with T cells respectively on the ratio of 1∶〖KG-*2〗100, 1∶〖KG-*2〗50 and 1∶〖KG-*2〗10. T cell proliferation stimulated by DC was determined using MTT method. DCs were co-cultured with T cells on the ratio of 1∶〖KG-*2〗10, and the supernatant were collected at 24 h, 48 h and 72 h. IFN-γ in the supernatant was detected with ELISA method.RESULTS: Induced by GM-CSF and IL-4, then stimulated by IL-18, TNF-α or IL-18+TNF-α, monocytes showed typical morphous of DC. No morphological difference was observed among DCs of the 3 groups. No statistical difference showed in expression level of CD1a, HLA-DR, CD83 and CD86 between IL-18 group and TNF-α group (P>0.05). The positive rates of CD1a and CD83 in IL-18+TNF-α group were higher than those in other 2 groups. The positive rate of HLA-DR in IL-18+TNF-α group was higher than that in IL-18 group. No difference between IL-18 group and TNF-α group in the potency of stimulating T cell proliferation was found, whereas the stimulating potency in IL-18+TNF-α group was higher than that in IL-18 group and TNF-α group. IL-12 in IL-18+TNF-α group at 48 h and 72 h was higher than that in IL-18 group and TNF-α group (P<0.05). However, there was no difference between the latter 2 groups. There was also no difference between IL-18 group and TNF-α group in IFN-γ secretion. IFN-γ in IL-18+TNF-α group was higher than that in IL-18 group and TNF-α group (P<0.05).CONCLUSION: Using IL-18 as intervent, DC expresses high level of surface molecules, secretes high level of IL-12, stimulates T cell proliferating effectively and produces IFN-γ potently. The actions are stronger when used in combination with TNF-α. It suggests that IL-18 may serve as a promoting agent of DC maturity, or combination with TNF-α in DC induction will strengthen the immune activity of DC.     

Liver X receptor activation and cholesterol efflux in macrophages from coronary atherosclerotic disease patients

AIM:To study the characteristic of liver X receptor alpha (LXRα), it’s target gene expression and cholesterol efflux from human macrophages in coronary atherosclerotic disease (CAD) patients. METHODS:Human monocyte-derived macrophages from CAD patients and controls were collected. Before being detected apoA-I-mediated human monocyte-derived macrophage cholesterol efflux and LXRα and mRNA expression of its target gene, the macrophages were induced with or without TO-901317. 〖JP2〗RESULTS:Compared with control normal macrophage, the mRNA levels of LXRα and its target gene expression were changed, and the macrophage cholesterol efflux was decreased in CAD patients. After stimulationwith TO-901317, the reactive capacity of LXR was also decreased from human monocyte-derived macrophage of CAD patients. CONCLUSION:The changes of cholesterol efflux and some gene expression in macrophages may be the pathogenetic cause atherosclerosis, and macrophage LXR activity may offer potential therapeutic benefit in the treatment of CAD.     

Relationship between SNP of TNFR gene and incidence/severity of pneumonia

AIM：To analyze the relationship between the single nucleotide polymorphism (SNP) of tumor necrosis factor receptor (TNFR) gene and the incidence and severity of pneumonia. METHODS：Total 132 Chinese individuals were enrolled in this study. There were 66 patients with pneumonia and 66 healthy subjects. The SNPs of TNFR gene including TNFR1+36A/G, TNFR1-609G/T, TNFR2+676T/G, TNFR2+1663T/G, TNFR2 +1668A/G and TNFR2 +1690C/T were genotyped by polymerase chain reaction-restriction fragment length polymorphism or gene sequencing for all subjects. Polymorphisms affecting pneumonia incidence and severity were calculated by SPSS. RESULTS：The frequencies of TNFR1-609G and T alleles in pneumonia patients were 40.9% and 59.1%, while those in healthy subjects were 53.8% and 46.2%. The frequency of TNFR1-609T in pneumonia patients was higher than that in healthy subjects (P<0.05). Besides, the frequencies of TNFR1-609G and T alleles in severe pneumonia patients were 25.0% and 75.0%, while those were 46.0% and 54.0% in non-severe pneumonia patients. The frequencies of TNFR2 +1690C and T alleles in severe pneumonia patients were 81.1% and 18.9%, while those were 61.0% and 39.0% in non-severe pneumonia patients. The frequencies of TNFR1-609T and TNFR2 +1690C in severity pneumonia subjects were higher than those in mild subjects (P<0.05). CONCLUSION：It appears that TNFR1-609T is associated with high incidence of pneumonia. TNFR1-609T and TNFR2+1690C are the risk factors of severity in pneumonia in Chinese.     

Effects of TNF-α-induced changes of expression and activity of 11-β HSD-1 on the insulin sensitivity in 3T3-L1 adipocytes

AIM: To observe the effects of tumor necrosis factor-α（TNF-α）-induced changes of expression and activity of 11-beta-hydroxysteroid dehydrogenase type1(11-β HSD-1) on the insulin sensitivity in 3T3-L1 adipocytes.METHODS: 3T3-L1 adipocytes were treated with TNF-α and TNF-α combined with aspirin, 2’-hydroxyflavanone or RU486, then mRNA expression and activity of 11-β HSD-1 and insulin-stimulated glucose uptake were examined.RESULTS: TNF-α increased expression and activity of 11-β HSD-1 in 3T3-L1 adipocytes, and decreased insulin-stimulated glucose uptake. Aspirin decreased expression and activity of 11-β HSD-1 induced by TNF-α, and alleviated the inhibiting effect of TNF-α on insulin-stimulated glucose uptake. 11-β HSD-1 specific inhibitor 2’-hydroxyflavanone and cortisol-receptor antagonist RU486 also alleviated the inhibiting effect of TNF-α on insulin-stimulated glucose uptake. CONCLUSION: TNF-α may decrease the insulin sensitivity in 3T3-L1 adipocytes through increasing expression and activity of 11-β HSD-1.     

Effects of ganmao shuangjie heji on the inflammatory damage in the lungs of mice infected by IV FM1

AIM: To establish the influenza infected mouse model and study the anti-inflammatory effect of ganmao shuangjie heji. METHODS: IV FM1 infected mice were used as the animal model. The changes of pathology and the cytokine TNF-α, IFN-γ and IL-10 in the lung were observed by HE staining and ELISA (double antibody sandwich enzyme linked immunosorbent assay) after ganmao shuangjie heji treatment. RESULTS: After infected by influenza virus, severe interstitial pneumonia was induced in the model group. Mild interstitial pneumonia was observed in ganmao shuangjie heji treated group. The protein expressions of cytokine TNF-α, IFN-γ and IL-10 were higher in model group than those in the control group. The protein expressions of TNF-α and IFN-γ in ganmao shuangjie heji treated group decreased and IL-10 expression increased significantly compared with model group. CONCLUSION: Ganmao shuangjie heji decreases the expressions of TNF-α and IFN-γ, and increases the expression of IL-10, thus, alleviates inflammatory injury. The clinical application of this medicine can shorten the course of disease.     

Effect of TNF-α on production and activation of caspase-3 in primary rat renal proximal tubule cells

AIM: To investigate the production and activation of caspase-3 in primary rat renal proximal tubule cells in response to tumor necrosis factor-α(TNF-α) and the implication of nuclear factor-κB (NF-κB) in the process. METHODS: Isolated rat renal proximal tubule cells (PTCs) from male adult Sprague Dawley rats were treated with TNF-α according to the indicated time courses. A specific NF-κB inhibitor, Bay11-7082, was used alone or as a pretreatment for 1 h followed by exposure to TNF-α for 24 h. The protein levels of cleaved caspase-3, caspase-3, I-κBα, phosphorylated I-κBα, and GAPDH were detected by Western blotting using specific antibodies. RESULTS: The protein level of cleaved caspase-3 relative to caspase-3 was significantly increased in the presence of TNF-α for 6 h, 12 h, and 24 h. Protein levels of caspase-3 were significantly decreased by 12 h and returned to baseline by 24 h in the presence of TNF-α. Treatment with Bay11-7082 for 25 h alone or pretreatment with Bay11-7082 for 1 h followed by addition of TNF-α for 24 h caused a remarkable reduction in both cleaved caspase-3 and caspase-3 as compared to control and TNF-α treated groups. An increase in phosphorylated I-κBα was observed from 15 min to 60 min after treatment with TNF-α at a dose of 10 μg/L in PTCs. CONCLUSION: NF-κB is not only associated with the activation of caspase-3 but also the production of caspase-3 in primary rat renal proximal tubule cells in response to TNF-α.     

Effect of inhibiting nuclear factor-kappa B on TNF-α-induced expression of angiotensinogen and production of angiotensinⅡ in glomerular mesangial cells

AIM: To study the effect of inhibiting nuclear factor-kappa B (NF-κB) activity on the expression of angiotensinogen (AGT) and the production of angiotensinⅡ (AngⅡ) induced by tumor necrosis factor-α (TNF-α) in glomerular mesangial cells (MCs) of SD rats. METHODS: The MCs of SD rats were isolated and divided into three groups as follows: control; MCs treated with TNF-α, and the MCs treated with TNF-α + pyrrolidinedithiocarbamate (PDTC). The activity of nuclear factor-kappa B was measured by electrophoretic mobility shift assay. The expression of AGT was determined by RT-PCR for mRNA and Western blotting for protein. The concentration of angiotensinⅡ in supernatant was measured by RIA. RESULTS: The NF-κB activity in the MCs treated with TNF-α (20.67±9.14)×102 μg/cell was significantly higher than that in control cells ［(8.25±4.35)×102 μg/cell, P<0.01］ and the MCs treated with TNF-α+PDTC ［(7.20±4.57)×102 μg/cell, P<0.01］, and no significant difference was found between control and the MCs treated with TNF-α＋PDTC (P>0.05). The AGT mRNA level in the MCs treated with TNF-α (0.27±0.05) was higher than that in the control cells (0.20±0.05, P<0.05), and no significant difference was observed when compared with that in the MCs treated with TNF-α＋PDTC (0.22±0.06, P>0.05). The expression of AGT protein in the MCs treated with TNF-α (0.60±0.19) μg/cell was higher than that in the control ［(0.37±0.15)μg/cell, P＜0.05］ and the MCs treated with TNF-α＋PDTC ［(0.37±0.17)μg/cell, P<0.05］, and no significance was found between the MCs treated TNF-α＋PDTC and the control (P>0.05). The AngⅡ level in supernatant of cultured MCs treated with TNF-α ［(9.73±2.38)×10-5 ng·L-1/cell］ was significantly higher than that in the control ［(7.50±1.51)×10-5 ng·L-1/cell, P<0.05］ and in the MCs treated with TNF-α＋PDTC ［(6.94±1.46)×10-5 ng·L-1/cell, P<0.05］, however, the difference between the MCs treated with TNF-α＋PDTC and the control was of no significance (P>0.05). CONCLUSION: TNF-α activates the NF-κB in glomerular MCs, induces the AGT expression and the production of AngⅡ. Inhibition of NF-κB decreases the AGT expression and the production of AngⅡ. Therefore, the effects of TNF-α on AGT and AngⅡ may be mediated by NF-κB.     

Expression of TNF-α and TGF-β in oral lichen planus patients

AIM: To investigate the expression of TNF-α， TGF-β in patients with oral lichen planus (OLP) and normal oral mucosa (NOM). METHODS: An immunohistochemical technique was performed to detect TNF-α, TGF-β1 and TGF-β2 expression in 22 cases with OLP and 10 normal controls. RESULTS: In lamina propria of OLP, the expression of TNF-α and TGF-β1 were increased, whereas TGF-β2 did changed significantly compared with control group. TNF-α positive signal were mostly found in macrophages, lymphocytes. TGF-β1 positive cell was present in macrophages, endothelial cells and fibrocytes. CONCLUSION: TNF-α and TGF-β1 play an important role in the development and maintenance of OLP local inflammation.     

Angiotensin Ⅱ stimulates TNF-α and NO production in peripheral blood mononuclear cells in heart failure patients

AIM: To examine the change of serum tumor necrosis factor-α (TNF-α), nitric oxide (NO) in patient with congestive heart failure (CHF) and the effect of angiotensin Ⅱ (AngⅡ), valsartan on TNF-α and NO production in culture peripheral blood mononuclear cells (PBMC), to assess the relationship between the renin-angiotensin system and cytokines. METHODS: Venous blood of both healthy volunteers (n=12) and patients with CHF (n=16) were collected. Serum TNF-α and NO were examined. Peripheral blood mononuclear cells (PBMC) were obtained from both the control and the patients groups and cultured with AngⅡ at concentrations of 0, 0.01, 0.1, 1 μmol/L, respectively. AngⅡ at concentration of 0.1 μmol/L combined with 0.1 μmol/L of valsartan was also used. After 24 h incubation, the contents of TNF-α and NO in the culture supernatants were measured. RESULTS: Serum TNF-α and NO production in CHF group were significantly higher than that in control group (P<0.01). The higher the heart failure degree, the higher the levels of TNF-α and NO (P<0.01), and no significant among different etiologies of CHF (P＞0.05) were observed. AngⅡ stimulated TNF-α and NO release from PBMC of patients with CHF and normal person, which was inhibited by valsartan. CONCLUSIONS: AngⅡ obviously increases TNF-α and NO production from PBMC, which indicates there is relationship between the renin-angiotensin system and TNF-α, NO. The fact that valsartan inhibits TNF-α production may be one of the mechanisms in treating CHF.     

Changes of arachidonic acid and TNF-α in primary glomerular disease

AIM: To explore the changes of arachidonic acid (AA) and tumor necrosis factor α(TNF-α) in primary glomerular disease and their effects on oxidative stress. METHODS: Fifty-five patients with primary glomerular disease confirmed by clinical examination and renal biopsy were selected. The patients were divided into 3 groups according to their glomerular filtration rate as chronic kidney disease(CKD)1,2 group, CKD3,4 group, and CKD5 group. The levels of free AA, TNF-α, malondialdehyde(MDA) and high-sensitive C-reactive protein(hs-CRP) in serum were detected by ELISA. RESULTS: No significant difference of AA and MDA between CKD1,2 group and CKD3,4 group was observed (P>0.05). The level of AA was obviously higher, and the level of MDA was significantly lower in CKD5 group than those in the other groups (P<0.05). No difference of TNF-α concentration between CKD3,4 group and CKD5 group was found (P>0.05), but that was significantly lower than that in CKD1,2 group (P<0.05). The content of hs-CRP in each group was not different (P>0.05). The level of MDA was negatively correlated with the level of AA (r=-0.752,P<0.01), whereas positively correlated with the level of TNF-α (r=0.463, P<0.01). The multiple linear regression equation of MDA(Y) for AA(X₁) and TNF-α(X₂) was Y=1 361.723-2.661X₁+9.320X₂ (F=52.445, P<0.01). CONCLUSION: AA and TNF-α are the important factors that influence the level of oxidative stress in primary glomerular disease. AA inhibits and TNF-α promotes oxidative stress.     

Effect of recombinant connective tissue growth factor on the phenotype transition of rat adventitia fibroblasts

AIM:To construct pcDNA3.1(+)/connective tissue growth factor (CTGF) eukaryotic expression plasmid and to investigate its role in the promotion of phenotypic transition in adventitia fibroblasts (AF). METHODS:The expression vector pcDNA3.1(+)/CTGF was constructed by routine molecular biological method. The expression vector pcDNA3.1(+)/CTGF was confirmed by restriction enzyme digestion and sequencing method. The expression vector pcDNA3.1(+)/CTGF was transfected into AF and the exogenous expression was observed. The expression of the α-SM 〖JP+1〗actin was examined by Western blotting. RESULTS:The eukaryotic expression vector of CTGF was successfully constructed, which was transfected into AF, the expressed CTGF promoted phenotype transition in AF. CONCLUSION:The pcDNA3.1(+)/CTGF plasmid was constructed and transfected into AF, the expressed CTGF promoted phenotype transition in AF.     

Effect of fucoidan on inhibition of proliferation and induction of apoptosis in human breast carcinoma cell line MCF-7

AIM: To investigate the effects and related mechanism of fucoidan on the proliferation and apoptosis in breast carcinoma cell line MCF-7. METHODS: MCF-7 cells were treated with different concentrations of fucoidan (100 mg/L, 300 mg/L, 500 mg/L, 1 000 mg/L) for 48 h. Cell viability was measured by MTT assay. Apoptosis morphological and biochemical changes were detected by Hoechst 33258 staining and agarose gel electrophoresis. The expression of 〖STBX〗bcl-2〖STBZ〗 and bax was examined by RT-PCR and Western blotting. RESULTS: Fucoidan at different concentrations (100 mg/L, 300 mg/L, 500 mg/L, 1 000 mg/L) effectively inhibited the proliferation of MCF-7 cells (P<0.01). The inhibitory ratio and apoptosis rate increased in a concentration-dependent manner. Agarose gel electrophoresis of DNA revealed the characteristic “ladder” pattern of apoptosis. Fucoidan down-regulated the expression of 〖STBX〗bcl-2〖STBZ〗 and up-regulated bax in the levels of mRNA and protein. The ratio of Bcl-2 to Bax decreased as the concentrations of fucoidan increased (P<0.05). CONCLUSION: Fucoidan inhibits the cell proliferation by inducing cell apoptosis, and the apoptosis is related to the down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of apoptotic protein Bax.     

SC58125 regulates TNF-α induced apoptosis in human colonic carcinoma cell line HT29 by inhibiting IκBα degrades

AIM:To investigate whether SC58125 synergizes with TNF-α to induce HT-29 cell apoptosis and study the possible molecular mechanism. METHODS:By using MTT,Hoechst 33342 staining and agarose gel electrophoresis,the effect of SC58125/TNF-α on the proliferation and apoptosis of HT-29 cell line was examined. The activity of caspase-3,the expression of IκBα,the phosphorylation level of IκBα,and the activation of NF-κB were measured after treatment with SC58125 by electrophoretic mobility shift assay and Western blotting.RESULTS:Both SC58125 and TNF-α exhibited cytotoxicity. The combination of the two reagents significantly reduced HT-29 cell viability in a dose-dependent manner. SC58125 and TNF-α co-treated cells showed induced nuclear condensation and fragmentation,and led to oligonucleosomal cleavage of genomic DNA,which was accompanied by the induction of caspase activity. IκBα levels were substantially decreased by the treatment of TNF-α. The degradation of IκBα was almost completely inhibited when SC58125 was added. NF-κB was activated in HT-29 cells after treatment with TNF-α,whereas pretreatment of HT-29 cells with SC58125 for 2 h,TNF-α induced NF-κB DNA binding was profoundly suppressed. CONCLUSION:SC58125 synergizes with TNF-α to inhibit cell growth and induces apoptosis in HT-29 cells,which may be mediated by activating caspase-3 and preventing degradation of IκBα.     

Effects of silymarin on LPS-induced acute lung injury in rats

AIM:To investigate the effects of silymarin on lipopolysaccharide (LPS)-induced acute lung injury in rats and its possible molecular mechanisms. METHODS:Fifty-eight male SD rats, weighting 230-250 g, were divided into four groups randomly: normal control (n=12); acute lung injury group (n=15), receiving intravenous LPS (O55∶〖KG-*2〗B5, 5 mg/kg); silymarin alone group (50 mg/kg, n=15); intervention group (n=16, receiving silymarin 50 mg/kg and LPS 5 mg/kg). The specimens were collected 6 hours later. The following changes, including blood gas analysis, the lung wet/dry weight ratio, the pulmonary vascular permeability, histological manifestations, lung tissue myeloperoxidase activity, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene in plasma and lung tissue, were observed. RESULTS:Compared with control group, the lungs of the rats in LPS treatment group showed significant hyperemia and spotted hemorrhage. The inflammatory granulocyte infiltrating, diffused alveolar septum thickening and spotted hemorrhage were observed in pathological examinations. The lung wet/dry weight ratio and Evans blue content (per gram) increased significantly after LPS treatment. The myeloperoxidase activity in plasma and lung tissue, the levels of TNF-α, IL-1β, MCP-1 and SOD, GSH-Px as well as malonaldehyde and conjugated diene were increased significantly in LPS treatment group. However, in intervention groups, all the above-mentioned measurements were reversed significantly by silymarin treatment compared with LPS treatment group. CONCLUSION:Silymarin may decrease inflammatory reaction and oxidative stress, and further decrease lung damage induced by LPS in rats, all indicating protection of silymarin against acute lung injury.     

Effects of curcumin on actinomycin D/TNF-α-induced injury in PC12 cells and rat hippocampal neurons

AIM: To study the possible mechanism of curcumin on actinomycin D (ActD)/tumor necrosis factor α (TNF-α)-induced injury in PC12 cells and rat hippocampal neurons. METHODS: PC12 cells were divided into control group, TNF-α group, ActD group, curcumin group, ActD/TNF-α group and curcumin+ActD/TNF-α group. The cells were cultured for 24 h. Inverted fluorescence microscopy was used to observe the morphological changes of the cells in each group. Annexin V/PI double staining was applied to analyze the apoptosis of PC12 cells. The level of intracellular Ca²⁺ was detected by Fluo-3 AM staining. Rat hippocampal slices were prepared and divided into the same groups as the PC12 cells. Extracellular microelectrode recording technique was used to observe and calculate the changes of long-term potentiation (LTP) in different groups. RESULTS: Apoptosis of PC12 cells was induced by ActD/TNF-α. Curcumin protected the PC12 cells from ActD/TNF-α-induced apoptosis (P<0.05). ActD/TNF-α increased the intracellular Ca²⁺ concentration. Curcumin significantly reduced ActD/TNF-α-induced apoptosis by decreasing the intracellular Ca²⁺ concentration (P<0.05), inversed the effect of ActD/TNF-α on LTP in hippocampal slices, and improved the synaptic plasticity (P<0.05). CONCLUSION: Curcumin protects ActD/TNF-α-induced neuronal damage by depressing the intracellular Ca²⁺ concentration and maintaining the homostasis of intracellular calcium.     

Association of TNF-α upregulation of MMP-9 activation in monocyte-derived macrophages with progression of joint damage in patients with rheumatoid arthritis

AIM: To explore the expression and activation of matrix metalloproteinase-9 (MMP-9) from monocyte-derived macrophages induced by tumor necrosis factor-α (TNF-α) and to investigate its association with progression of joint damage in patients with rheumatoid arthritis. METHODS: TNF-α and MMP-9 in serum and synovial fluid from patients with early RA and controls were tested with a double-antibody enzyme-linked immunosorbent assay. The correlation between MMP-9 and Larsen score over the first 12 months was analyzed. THP-1 cells differentiated by the treatment with TPA were stimulated with increasing concentration of TNF-α for 24 h in vitro. The protein expression of MMP-9 was determined by Western blotting. The activity of MMP-9 was measured by gelatinolytic zymography. Boyden chamber-matrigel in vitro invasion assay was used to detect the invasive capacity. RESULTS: The levels of TNF-α and MMP-9 in serum and synovial fluid of RA patients were significantly higher than those in controls (P<0.05). Serum and synovial fluid levels of MMP-9 correlated significantly with Larsen score (r=0.37 and 0.32, P<0.01). The MMP-9 activity and invasive ability of co-cultured THP-1 cells with TNF-α and TPA were higher than those of non-TNF-α treatment. CONCLUSION: TNF-α upregulates MMP-9 activation and promotes infiltration of monocyte-derived macrophages, indicating that TNF-α play an important role in the pathogenesis of RA.     

Effects of ouercetin on lipopolysaccharide induced hepatocyte injury in vitro

AIM: To study the effects of quercetin on lipopolysaccharide (LPS) induced hepatocyte injury and the expression of TNF-α in vitro. METHODS: Hepatocytes were isolated from male Sprague-Dawley rats by collagenase perfusion. LPS at concentration of 40 mg/L was used to induce injury to the cultured cells, and 0.5-10 μmol/L quercetin was added at the same time. After 24 h of incubation, the cell apoptosis rates were detected by MTT and PI-AnnexinV. LDH and TNF-α were measured by kits. RESULTS: 40 mg/L LPS caused a 27% growth inhibition. The apoptosis rate was 30.2%. LDH leakage was 20 folds higher than normal. TNF-α expression significantly increased. Treated with quercetin at doses of 0.5-10 μmol/L, the apoptosis rate, LDH leakage and TNF-α expression in hepatocytes were attenuated in a dose dependent manner. CONCLUSION: 0.5-10 μmol/L of quercetin protects hepatocytes from injury induced by LPS, which is associated with suppression of the inflammatory cytokine TNF-α.     

Protective effects and mechanism of astragalus injection on asthmatic rats

AIM: To explore the protective effects and mechanism of astragalus injection on asthmatic rats.METHODS: OVA was injected intraperitoneally and inhaled to produce the asthmatic model.Forty rats were randomly divided into five groups: control group,asthma group and astragalus groups of high,medium and low dose.The concentrations of IL-4,IFN-γ in BALF,the expression of IL-4 mRNA,IFN-γ mRNA and phospho-p38 MAPK in lung tissues were respectively measured by ELISA,RT-PCR and Western blotting.The number of inflammatory cells in BALF and histropathology changes were observed.RESULTS: In asthmatic group,the number of inflammatory cells and the concentrations of IL-4 in BALF and the expression of IL-4 mRNA,phospho-p38 MAPK in lung tissue were higher,but IFN-γ and IFN-γ mRNA were lower than those in normal control rats (P<0.01).In astragalus group,the number of inflammatory cells,the concentrations of IL-4 in BALF and the expression of IL-4 mRNA,phospho-p38 MAPK in lung tissue were lower,but IFN-γ and IFN-γ mRNA were higher than those in normal control rats (P<0.01),and histropathology damage was alleviated significantly.The efficacies in the astragalus groups of high,medium and low dose were similar,which no significant difference was observed among them.There were positive correlations between the expression of 〖JP3〗phospho-p38 MAPK and the number of eosinophil,the concentration of IL-4,IL-4 mRNA (r=0.63,r=0.69,r=0.71,〖JP〗 P<0.01),and negative correlations between the expression of phospho-p38 MAPK and IFN-γ and IFN-γ mRNA (r=-0.65,r=-0.68,P<0.01).CONCLUSION: p38 MAPK may play a role in pathological process of asthma.Astragalus effectively treats asthma by inhibiting the expression of phospho-p38 MAPK,correcting the inbalance of IFN-γ/ IL-4 and decreasing the number of inflammatory cells.