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Aim To investigate the effect of phillygenin ( PHI) on lipopolysacchride ( LPS) and normal human plasma ( NHP) induced inflammatory injury on alveolar type II epithelial A549 cells and the related mechanism. Methods A549 cells were exposured to 1 mg • L
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Aim To investigate the hepatotoxicity of rutecarpine(RUT)by using high-content screening technology.Methods HepG2 cells were exposed to different concentrations of RUT for different time, then cell viability was detected by MTT method.Cell count, nucleus injury, mitochondrial membrane potential(MMP), reactive oxygen species(ROS), internal flow of calcium, cell membrane integrity(DIR)were measured by high-content screening technology.The activation of MAPK, NF-κB and JAKs-STATs was assayed by high-content screening technology.The apoptosis was detected by flow cytometry.Results The viability was significantly reduced by 100 μmol·L-1 RUT(P<0.01)after HepG2 cell exposure to RUT for 24 h, the nuclear area decreased and the nuclear morphology was uneven, and after 48 h, the cell count was significantly reduced(P<0.01), the early apoptosis was detected(P<0.01).After HepG2 cell exposure to RUT for 6 h, the levels of ROS and internal flow of calcium significantly increased(P<0.01), and the cell membrane integrity was obviously damaged(P<0.01).After exposure to 100 μmol·L-1 RUT for 24 h, the phosphorylation of ERK, JNK, STAT3 and p38 significantly increased(P<0.01, P<0.05), but there was no significant change in total protein level.The expression of c-Jun and c-Fos was up-regulated at 3 h(P<0.01), and at 3h time point, the phosphorylation of NF-κB p65 significantly increased(P<0.01), but nuclear translocation was not significant.Conclusions Under certain conditions, RUT shows cytotoxicity on HepG2 cells, and its toxic mechanism is mainly related to injury caused by oxidative stress and inflammatory response.
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Aim To study the effect of proliferation and activation of vascular smooth muscle cells (VSMCs) induced by activated the complement alternative pathway and intervention. Methods Normal human plasma was specifically activated the complement alternative pathway by incubated with cobra venom factor (CVF). Exposed VSMCs to the activated complement products, the change of cell morphology and the cell viability were assayed by inverted phase contrast microscope and MTT method, respectively. The supernatant was assayed for expression of E-selectin, ICAM-1 and VCAM-1 by using ELISA reagent kits. The protein expression levels of p-NF-kB p65, NF-kB p65 and IKK were assayed by Western blot. The nucleus transcriptional activity of NF-ΚB p65 was tested by the dual luciferase reporter assay system. Pyrrolidine dithiocarbamate (PDTC) was used to intervene the proliferation and activation of VSMCs. Results VSMCs were activated and induced to proliferation after exposed to the products of activated complement alternative pathway. The expressions of E-selectin, VCAM-1 and ICAM-1 were up-regulated. The contents of ICAM-1 and VCAM-1 reached the peak at 6 h and the E-selectin increased significantly at 12 h. Meanwhile, the phosphorylation level of NF-ΚB p65, nucleus transcriptional activity of NF-ΚB p65 and the protein expression of IKK and N F - Κ B p65 increased. The above mentioned changes were clearly inhibited by PDTC. Conclusions The activated complement alternative pathway can initiate proliferation and activation of VSMCs, and its mechanism goes hand in hand with activation of N F - Κ B signaling pathway. PDTC, a specific inhibitor of N F - K B, can effectively inhibit the proliferation and activation of VSMCs.
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Aim To investigate the protective effect of baicalin on inflammatory response of human microvascular endothelial cells ( HMECs) induced by lipopo- lysaccharides ( LPS) and its mechanism. Methods LPS was applied to establish inflammatory model on HMECs in this work. HMECs were pretreated with different concentrations of baicalin ( 1. 0 x 10 "6 , 1. 0 x 10 ~7 and 1. 0 x 10 "8 mol • L"1) , and then exposed to LPS. The supernatant was removed and assayed for expression of the adhesion molecule ICAM-1, the inflam- atory mediator IL-6 and MCP-1 by using ELISA reagent kits. The inward flow of calcium ion was observed by fluorescence microscope, and the intracellular calcium ion level was measured by flow cytometry. The fluorescence intensity of nucleus NF-kB p65 was detected by immunoflourescent technique. The nucleus transcriptional activity of NF-kB was measured by the dual lu- ciferase reporter assay system. Moreover, the expression of protein NF-kB p65, phospo-NF-kb p65 ( p p65 ) and Toll like receptor 4 (TLR4 ) were detected by Western blot. Results The internal flow of calcium, the phosphorylation of NF-kB p65 and the transcription activity significantly increased, and the expression of ICAM-1, IL-6 and MCP-1 up-regulated after HMECs were exposed to LPS. Baicalin inhibited the inward flow of calcium ion, the nuclear translocation of NF-kB p65 and the up-regulation of transcriptional activity, decreased the expression of ICAM-1, IL-6 and MCP-1, and down-regulated the expression of NF-kB p65, p- p65 and TLR4 in a dose-dependent manner. Conclusions Baicalin possesses a protective effect on inflammatory response of endothelial cells induced by LPS, and its mechanism may be highly related to the inhibition of the internal flow of calcium and the activation of NF-kB signaling pathway, and thus reduce the level of inflammatory response.
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Aim To explore the anticomplementary ac-tivity and active constituents of Phyllanthus urinaria. Methods Being guided by bioactive screening,the anticomplementary constituents of P.urinaria were iso-lated and purified by solution partition and chromato-graphic techniques,and their structures were identified by 13C-NMR spectrum and the comparison of reported data. The anticomplementary activity and possible mechanism were assayed.Results The EtOAc fraction of the methanol extract of P.urinaria was shown to be the active fraction,and two compounds were purified from the fraction and were identified as corilagin and ellagic acid.The EtOAc and the two compounds signifi-cantly inhibited the hemolysis of the complement classi-cal pathway,with IC50values of 53.77 mg·L-1, 176.54 mg·L-1and 102.23 mg·L-1,respectively. While they just showed slight effect on inhibiting the hemolysis of the complement alternative pathway. All of them affected the formation of C3 convertase of the classical pathway. Conclusions The polyphenols are main anticomplentary constituents of P. urinaria,and its mechanism is related to inhibiting formation of C3 convertase of the classical pathway.
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Aim To explore the protective effect and mechanism of icariin ( ICA ) on acute lung injury ( ALI) in mice induced by activation of the comple-ment alternative pathway. Methods 32 healthy KM mice were randomly divided into four groups: the nor-mal group, the model group, the PDTC group and the Icariin group, which received 7-day intragastric admin-istration respectively. Then cobra venom factor ( CVF) was used to activate specifically complement alternative pathway to induce acute lung injury in mice by intrave-nous injection. Myeloperoxidase ( MPO ) activity of lung homogenate, the cell count and the protein con- tent of bronchoalveolar lavage fluid ( BALF ) were measured. The concentration of IL-6, TNF-α, P-selec-tin and ICAM-1 in BALF and serum were determined by ELISA. The pathological change of lung tissue was observed by HE staining. The phosphorylation of NF-κB p65 in lung tissues was checked by immunohisto-chemistry. The effect of the transcriptional activity of NF-κB signal pathway in microvascular endothelial cells was measured by employing dual-luciferase re-porter assay system. Results ICA reduced MPO ac-tivity of lung homogenate, the cell count and the con-tent of IL-6, TNF-α, P-selectin in BALF obviously. The level of TNF-α, P-selectin and ICAM-1 in serum was decreased, the pulmonary inflammatory cell infil-tration was reduced, the phosphorylation of NF-κB p65 in lung was inhibited significantly and the transcrip-tional activity of NF-κB was also down-regulated. Con-clusion ICA can alleviate acute inflammatory re-sponse of ALI mice induced by activation of the com-plement alternative pathway. The mechanism may be highly related to the inhibition of inflammatory cell in-filtration in lung tissue, the down-regulation of phos-phorylation of NF-κB p65 and nuclear transcriptional activity.
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<p><b>BACKGROUND</b>It is widely accepted that tumor necrosis factor-α (TNF-α) plays an important role in the pathogenesis of emphysema. This study aimed at investigating the protective effects of anti-TNF-α antibody, infliximab, in the development of emphysema induced by passive smoking in rats.</p><p><b>METHODS</b>Thirty-nine rats were randomly divided into a normal control group (group 1), an emphysema group (group 2), and an infliximab-intervention group (group 3). Rat models of emphysema were established by exposure to cigarette smoking daily for 74 days. After 1 month, the infliximab intervention group was treated with infliximab via subcutaneous injection. The levels of TNF-α, IL-8 and vascular endothelial growth factor (VEGF) in bronchoalveolar lavage fluid (BALF) were measured with enzyme linked immunosorbent assay (ELISA). The number and classification of cells in the BALF were measured. Lung tissue sections stained by hematoxylin and eosin (HE) were observed, and mean linear intercept (MLI) and mean alveolar numbers (MAN) were measured. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods were used to examine the percentage of positive cells and distribution of apoptotic cells.</p><p><b>RESULTS</b>The levels of TNF-α and IL-8 in BALF were higher in group 2 than in group 1 and group 3. The MLI was greater in group 2 than that in group 1 and group 3 while MAN was decreased. The concentration of VEGF in BALF of group 2 was significantly decreased as compared with group 1. The total cells and neutrophils number was significantly increased in group 2 as compared with group 1 and group 3, so was the percentage of neutrophils. The number of TUNEL positive cells in the alveolar septa was significantly increased in group 2 as compared with group 1 and group 3.</p><p><b>CONCLUSION</b>Infliximab protects against cigarette smoking-induced emphysema by reducing airway inflammation, attenuating alveolar septa cell apoptosis and improving pathological changes.</p>
الموضوعات
Animals , Male , Rats , Antibodies, Monoclonal , Therapeutic Uses , Bronchoalveolar Lavage Fluid , Chemistry , Cell Biology , Infliximab , Interleukin-8 , Metabolism , Pulmonary Alveoli , Cell Biology , Pulmonary Emphysema , Metabolism , Random Allocation , Rats, Sprague-Dawley , Tobacco Smoke Pollution , Tumor Necrosis Factor-alpha , Metabolismالملخص
Thirteen compounds from Dendrolobium triangulare (Retz.) Schindl. were isolated and purified by chromatography on silica gel, macroporous resin column and recrystallization method, and their structures were elucidated by chemical and spectral analyses as azo-2, 2'-bis [Z-(2, 3-dihydroxy-4-methyl-5-methoxy) phenyl ethylene] (1), beta-sitosterol (2), N-(2'-hydroxy-tetracosanoyl)-2-amino-1, 3, 4-trihydroxyoctadec-8E-ene (3), lupeol (4), cycloeucalenol (5), daucosterol (6), betulinc acid (7), betulin (8), glyceryl hexacosanoate (9), glyceryl 26-hydroxy hexacosanoate (10), methyl pheophorbide-a (11), acacetin-7-O-alpha-L-rhamnopyranosyl (1-6)-beta-D-glucopyranoside (12) and robinin (13). To our knowledge, all compounds are obtained from Dendrolobium genus for the first time and compound 1 is a novel compound. Moreover, it is understood that compound 1 has better protection against PC12 cell damnification deduced by glutamate, than that of Vitamin E in 2 microg x mL(-1) concentration.
الموضوعات
Animals , Rats , Azo Compounds , Pharmacology , Fabaceae , Chemistry , PC12 Cellsالملخص
<p><b>AIM</b>Nineteen compounds related to salicylic acid were evaluated for their in vitro activity of inhibiting beta-lactamase isolated from a resistant strain of Pseudomonas aeruginosa, and their structure-activity relationships were examined.</p><p><b>METHODS</b>Nitrocefin method was used.</p><p><b>RESULTS</b>The 50% inhibitory concentration (IC50) of salicylic acid inhibiting beta-lactamase was 22 mmol x L(-1); four analogs had IC50 lower than that of salicylic acid; fifteen analogs had IC50 higher than that of salicylic acid.</p><p><b>CONCLUSION</b>Examination of the structure-activity relationships of the compounds revealed that carboxyl group and adjoining hydroxyl group were active group, and replacement of adjoining hydroxyl by carboxyl increased activity nearly 4-fold. Moreover, addition of a sulfonic group at C-5 and nitro group at C-3, 5 of benzenoic ring of salicylic acid resulted in a 2-fold to 3-fold increase in activity, addition of a amino group at C-5 of benzenoic ring of salicylic acid decreased activity, add addition of -Cl or -F at C-2,4 position of benzenoic ring of benzoic acid did not show activity.</p>
الموضوعات
Anti-Bacterial Agents , Chemistry , Pharmacology , Cephalosporins , Metabolism , Inhibitory Concentration 50 , Pseudomonas aeruginosa , Salicylates , Chemistry , Pharmacology , Structure-Activity Relationship , beta-Lactamases , Metabolismالملخص
<p><b>OBJECTIVE</b>To study chemical constituents of Incarvillea arguta and their accelerating PC-12 cell differentiation.</p><p><b>METHOD</b>The constituents were isolated and repeatedly purified on silica gel column chromatography, and were identified on the basis of physicochemical and spectroscopic analysis. The neurotrophic activity of different portion and all purified compounds from I. arguta was determined on the model of PC-12 cell.</p><p><b>RESULT</b>Five compounds were isolated from BuOH portion of alcohol extraction of I. arguta. Their structures were identified as plantarenaloside (I), 5-hydroxy-4', 6 7-trimethoxy-flavone (II), 4', 5-dihydroxy-6, 7-dimethoxyflavone (III), 4', 5-dihydroxy-7-methoxyflavone (IV), 5-dydroxy-4', 7-dimethoxyflavone (V).</p><p><b>CONCLUSION</b>Compound I is isolated from the plant for the first time and it has neurotrophic activity for PC-12 cell. Compounds II approximately V are isolated from the genus Incarvillea for the first time.</p>