This study explored the curative effect of Jingfang Mixture on urticaria mice induced by aluminum hydroxide/ovalbumin, and discussed its mechanism. Sixty male Kunming mice were randomly divided into a normal group, a model group, three Jingfang Mixture(low-dose, medium-dose, and high-dose) groups, and a positive drug(cetirizine hydrochloride) group. The urticarial model in mice was induced by the intraperitoneal injection of the mixed solution of ovalbumin and aluminum hydroxide. The degrees of pruritus were observed after the second immunization. Pathological changes were detected by hematoxylin and eosin(HE) staining. Levels of interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α) in the serum were detected by enzyme linked immunosorbent assay(ELISA). Expressions of NOD-like receptor protein 3(NLRP3) and IL-1β were detected by immunohistochemistry(IHC). Expressions of nuclear factor kappa-B(NF-κB p65), NLRP3, apoptosis-associated speck-like protein containing a CARD(ASC), cysteinyl aspartate-specific proteases 1(caspase-1), and IL-1β proteins were detected by Western blot. The results showed that, except for the normal group, the mice in all groups had different degrees of pruritus. Compared with the model group, the Jingfang Mixture groups and the positive drug group prolonged the scratching latency of mice(P<0.05), and significantly reduced the number of scratching(P<0.05). In addition, the Jingfang Mixture groups and the positive drug group improved the pathological morphology of skin tissue. The expression levels of IL-1β and TNF-α in serum were significantly reduced(P<0.05), and the number of NLRP3 and IL-1β positive cells was decreased(P<0.01). The expressions of p-NF-κB p65, NLRP3, ASC, cleaved caspase-1, and IL-1β protein were significantly down-regulated(P<0.05). The results of the above study indicate that Jingfang Mixture inhibit the inflammatory response in urticaria mice, and the mechanism may be related to the inhibition of activating NF-κB/NLRP3/IL-1β signaling pathway.
Animals ; Male ; Mice ; NF-kappa B/metabolism* ; Interleukin-1beta/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Ovalbumin ; Aluminum Hydroxide/pharmacology* ; Signal Transduction ; Caspase 1/metabolism* ; Urticaria ; Pruritus

OBJECTIVETo explore the effect of aluminum phosphate gel and Kangfuxin on esophageal pathology and expressions of interleukin-8 (IL-8) and prostaglandin E2 (PGE2) in rats with reflux esophagitis and explore the possible mechanisms.

METHODSSixty SD rats were randomized into aluminum phosphate gel group (n=10), Kangfuxin group (n=10), aluminum phosphate gel+Kangfuxin group (n=10), model group (n=20), and control group (n=10). Except for those in the control group, all the rats were subjected to infusion of diluted lysolecithin with hydrochloric acid in the esophagus for 14 days. Ten rats in the model group and those in the control group were sacrificed to examine the pathological changes and contents of IL-8 and PGE2 in the esophagus using optical and electron microscopes and radioimmunoassay. The next day the rest rats were given corresponding treatments (saline in model group) administered into the esophagus on a daily basis for 14 days, after which esophageal pathologies and IL-8 and PGE2 contents were examined.

RESULTSThe model rats showed obvious esophageal pathologies including inflammatory cell infiltration, vacuolar degeneration of the epithelial cells, esophageal erosion and even ulceration, with severe detachment of the epithelial cells. The rats in all the intervention groups showed lessened esophageal pathologies and lowered esophageal IL-8 and PGE2 contents compared with those in the model group. Esophageal mucosal injury index and IL-8 and PGE2 contents were all significantly lower in rats receiving combined treatment with aluminum phosphate and Kangfuxin than in those receiving either of the treatments (P<0.05).

CONCLUSIONSBoth Kangfuxin and aluminum phosphate gel are effective in the treatment for reflux esophagitis induced by lysolecithin and hydrochloric acid, and their therapeutic effects are achieved possibly by reducing IL-8 and PGE2 levels in the esophagus.

Aluminum Compounds ; pharmacology ; Animals ; Dinoprostone ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Esophagitis, Peptic ; drug therapy ; metabolism ; Esophagus ; drug effects ; pathology ; Gels ; Interleukin-8 ; metabolism ; Phosphates ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the influence of aluminum hydroxide adjuvant on a murine model of allergic rhinitis (AR) and to confirm an appropriate method of establishing a mouse model of AR. METHOD: Establishing two types of BALB/c mice models of AR, one was identified as Local group which was characterized through intranasal sensitization and challenge using ovalbumin (OVA), and the other Systemic group which was made by intraperitoneal sensitization with OVA plus aluminum hydroxide and intranasal challenge through OVA. Then the numbers of sneezing and nasal rubbing were counted after the last challenge and the eosinophils in the nasal mucosa of mice models were observed and counted though Luna stain. Furthermore, morphological hyperplasia was examined in intraepithelial goblet cells and submucosal glands with HE stain. In addition, interlukin (IL) -4, IL-5, OVA specific IgE (sIgE) and interferon (IFN)-gamma in nasal lavage fluid (NLF) and serum of mice were examined u sing enzyme linked immunosorbent assay (ELISA). RESULT: The counts of sneezing and nasal rubbing in local group were more than those in systemic group and eosinophilia in the nasal mucosa of former group was greater than that in the latter one. Morphological hyperplasia was stronger in intraepithelial goblet cells and submucosal glands in local group compared with that in systemic group. Furthermore, the contents of IL-4, IL-5 and sIgE increased in the NLF and serum of mice of local group compared to those of systemic one. However, the production of IFN-gamma of mice in local group decreased when compared with that in Systemic group. CONCLUSION OVA plus aluminum hydroxide adjuvant may promote Th1 type immune response as well as Th2 response. OVA intranasal sensitization and challenge locally is an appropriate way in the establishment of AR mice models.
Adjuvants, Immunologic ; pharmacology ; Aluminum Hydroxide ; pharmacology ; Animals ; Disease Models, Animal ; Immunoglobulin E ; immunology ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Interleukin-5 ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Rhinitis, Allergic ; immunology

OBJECTIVETo study synthesis of baicalin-copper and baicalin-aluminium complex and its antimicrobial, anti-tumor activity and anti-tumor effect against macrophages.

METHODBaicalin was reacted with metallic salt under a weak base condition to produce baicalin-copper and baicalin-aluminium complex. Baicalin and its synthesized complex were detected for antimicrobial activity against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans by twofold broth dilution technique. Their anti-tumor activity against A549 and IC50 of HepG2 cells and anti-tumor effect against macrophages were detected by the MTT. And their phagocytic effect on macrophages was determined by the neutral red assay.

RESULTThe yields of baicalin-copper and baicalin-aluminium complex were 73.93% and 91.08%, respectively. The minimum inhibitory concentration (MIC) value against Staphylococcus aureus, Hay bacillus, Escherichia coli, Salmonella and Candida albicans was 0.0004, 0.0009, 0.0004, 0.0009, 0.000 4 mol x L(-1) for baicalin-copper complex and 0.0011, 0.0011, 0.0011, 0.0011, 0.0005 mol x L(-1) for baicalin-aluminium complex. The IC50 values against A549 and HepG2 cells were 89.6, 22.6 micromol x L(-1) for baicalin-copper complex, and 138.8, 97.2 micromol x L(-1) for baicalin-aluminium complex. The inhibitory ratio of macrophage on A549 cell was 43.52%, 80.89%, 52.66%, respectively, after the macrophages were stimulated by baicalin, baicalin-copper and baicalin-aluminium complex at a concentration of 160 micromol x L(-1).

CONCLUSIONThe acute toxicity test in mice showed that the complex was nontoxic to mice. Baicalin-copper complex showed the highest antimicrobial, anti-tumor activity, and the strongest effect on the anti-tumor activity of macrophage, while baicalin showed the lowest activities compared with baicalin-copper and baicalin-aluminium complex.

Aluminum ; Animals ; Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Copper ; Drugs, Chinese Herbal ; chemical synthesis ; chemistry ; pharmacology ; Flavonoids ; Humans ; Mice ; Microbial Sensitivity Tests

In order to study the effect of seed soaking with different aluminum solution on seed germination and seedling physiological characteristics of Platycondon grandiflorum, two P. grandiflorum varieties'seed (the white flower and the purple flower) were soaked in Al3+ solution with different concentrations (0, 10, 100, 250, 500, 750 and 1000 mg x L) for 24 h, then germinated in illumination incubator. Results showed that the aluminum toxicity on the trends of the germination rate, germination index and vigor index was positive associated with its concentration, and the Al tolerance of the purple was slightly greater than that of the white. There were some relationships between the physiological indices, which were the leakage rate of electrolyte, the malonaldehyde (MDA) content, the activities of peroxidase (POD) and superoxide dismutase (SOD) , the free praline(Pro) and the soluble sugar contents, with the concentrations of Al. It was suggested that there was Al tolerance difference between the two P. grandiflorum varieties: the purple flower was greater than the white.
Aluminum ; pharmacology ; Dose-Response Relationship, Drug ; Germination ; drug effects ; Immersion ; Platycodon ; drug effects ; growth & development ; physiology ; Seedlings ; drug effects ; physiology ; Seeds ; drug effects ; growth & development

OBJECTIVETo study the effect of necrostatin (Nec-1) on apoptosis induced by aluminum (Al), and approach the mechanism.

METHODSNeural cell death model was made by 4 mmol/L Al treated neuroblastoma cells (SH-SY5Y). Cell viabilities were detected at different concentrations of Al and/or Nec-1. Hoechst 33342/PI double staining was used to observe apoptosis and (or) necrosis that were quantified by flow cytometry using Annexin V/PI double staining. Apoptotic pathway was tested by activities of Caspase-3, Caspase-8 and Caspase-9. In addition, the expression of NF-kappa B and Cyt-c was measured by immunocytochemistry.

RESULTSCell viabilities were significantly decreased with the increasing concentrations of Al (P < 0.05), which could be significantly upregulated by 60 micromol/L Nec-1 (P < 0.05) and were correlated with the concentrations of Nec-1 (P < 0.05, P < 0.01). Apoptosis and necrosis were observed under fluorescent microscope and quantified by flow cytometry, which suggested an increasing trend of apoptotic and necrotic rates (P < 0.05, P < 0.01). Whereas, Nec-1 could not only decrease the necrotic rate but also apoptotic rate as well (P < 0.05, P < 0.01). Data of Nec-1 on caspases activities showed that Nec-1 could not affect Caspase-9 activity (P > 0.05) and Cty-c protein expression as well (P > 0.05). However, Nec-1 could reduce Caspase-8 activity significantly (P < 0.05, P < 0.01) and increase NF-kappa B protein expression (P < 0.05, P < 0.01) and finally decrease Caspase-3 activity (P < 0.05).

CONCLUSIONNec-1 could reduce cell apoptosis induced by Al, through Caspase-8 pathway, and up-regulate the expression of NF-kappa B protein.

Aluminum ; toxicity ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Death ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cytochromes c ; metabolism ; Humans ; Imidazoles ; pharmacology ; Indoles ; pharmacology ; NF-kappa B ; metabolism ; Neuroblastoma

The present experiments were done to determine the effectiveness of a non-specific nitric oxide synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME), on oxidative stress parameters induced by aluminium chloride (AlCl3) intrahippocampal injections in Wistar rats. Animals were sacrificed 3 h and 30 d after treatments, heads were immediately frozen in liquid nitrogen and forebrain cortices were removed. Crude mitochondrial fraction preparations of forebrain cortices were used for the biochemical analyses: nitrite levels, superoxide production, malondialdehyde concentrations, superoxide dismutase (SOD) activities and reduced glutathione contents. AlCl3 injection resulted in increased nitrite concentrations, superoxide anion production, malondialdehyde concentrations and reduced glutathione contents in the forebrain cortex, suggesting that AlCl3 exposure promoted oxidative stress in this brain structure. The biochemical changes observed in neuronal tissues showed that aluminium acted as a pro-oxidant. However, the non-specific nitric oxide synthase (NOS) inhibitor, L-NAME, exerted anti-oxidant actions in AlCl3-treated animals. These results revealed that NO-mediated neurotoxicity due to intrahippocampal AlCl3 injection spread temporally and spatially to the forebrain cortex, and suggested a potentially neuroprotective effect for L-NAME.
Aluminum Compounds/*toxicity ; Animals ; Chlorides/*toxicity ; Glutathione/metabolism ; Male ; Malondialdehyde ; NG-Nitroarginine Methyl Ester/*pharmacology ; Nitric Oxide Synthase/*antagonists & inhibitors ; Nitrites/chemistry/metabolism ; Prosencephalon/*drug effects ; Rats ; Rats, Wistar ; Superoxide Dismutase/metabolism ; Superoxides/metabolism

OBJECTIVETo study whether necroptosis exists or not in neural cell death induced by aluminum.

METHODSSH-SY5Y cells were treated with 4 mmol/L AlCl(3) x 6H(2)O The cell viability was determined with CCK-8 kit after treated with Nec-1 at different dosages (0, 30, 60, 90 micromol/L). Mitochondria membrane potential (MMP), content of reactive oxygen species (ROS), and apoptotic rate/necrotic rates were measured with cytometry.

RESULTSNec-1 ameliorated the necrotic-like cell morphology, the cell viability were 0.28 +/- 0.05, 0.58 +/- 0.03, 0.68 +/- 0.04, and 1.03 +/- 0.17, there were significant differences between the Nec-1 treated groups and that of controls (t values were 3.25, 3.36, 4.56; P < 0.05). After Nec-1 treatment, the necrotic rates were 16.46% +/- 0.54%, 10.40% +/- 0.64%, 5.43% +/- 0.68%, and 6.28% +/- 0.35%, there were significant differences between the Nec-1 treated cells and that of controls (t values were 3.62, 7.32, 6.96; P < 0.05); while the apoptotic rates were 8.68 +/- 0.36, 7.66 +/- 0.53, 5.68 +/- 0.41, and 4.13 +/- 0.41, there was no significant difference among the groups (F = 6.33, P = 0.11). Cytometry had shown the increased cell MMPs after Nec-1 treatment, which were 67.54 +/- 6.36, 49.42 +/- 5.96, 84.79 +/- 6.86, and 95.51 +/- 7.01, there were significant differences as comparing MMPs of the middle and high dosage of Nec-1 treated cells with those of controls (t values were 3.21, 4.01; P < 0.05); while ROS contents in the Nec-1 treated SH-SY5Y cells were 54.07 +/- 3.32, 52.79 +/- 2.36, 54.68 +/- 1.91, and 59.23 +/- 2.96, there was no significant difference among the groups (F = 5.26, P = 0.19).

CONCLUSIONNec-1, as a specific inhibitor of necroptosis, might effectively block the cell death pathway induced by aluminum, it indicates that necroptosis should be one of the major causes of the SH-SY5Y cell toxicity induced by aluminum, and necroptosis also plays an important role in aluminum induced SH-SY5Y cell death.

Aluminum ; toxicity ; Apoptosis ; drug effects ; Cell Death ; drug effects ; Cell Line, Tumor ; Humans ; Imidazoles ; pharmacology ; Indoles ; pharmacology ; Neuroblastoma

BACKGROUNDIncreasing evidence suggests that many neurons may die through apoptosis in Alzheimer's disease (AD). Mitochondrial dysfunction has been implicated in this process of neuronal cell death. One promising approach for preventing AD is based upon anti-apoptosis to decrease death of nerve cells. In this study, we observed the memory improving properties of curcumin in mice and investigated the neuroprotective effect of curcumin in vitro and in vivo.

METHODSThe mice were given AlCl(3) orally and injections of D-galactose intraperitoneally for 90 days to establish the AD animal model. From day 45, the curcumin group was treated with curcumin for 45 days. Subsequently, the step-through test, neuropathological changes in the hippocampus and the expression of Bax and Bcl-2 were carried out to evaluate the effect of curcumin on the AD model mice. In cultured PC12 cells, AlCl(3) exposure induced apoptosis. The MTT assay was used to measure cell viabilities; flow cytometric analysis to survey the rate of cell apoptosis; DNA-binding fluorochrome Hoechst 33258 to observe nuclei changes in apoptotic cells and Western blot analysis of Bax, Bcl-2 to investigate the mechanisms by which curcumin protects cells from toxicity.

RESULTSCurcumin significantly improved the memory ability of AD mice in the step-through test, as indicated by the reduced number of step-through errors (P < 0.05) and prolonged step-through latency (P < 0.05). Curcumin also attenuated the neuropathological changes in the hippocampus and inhibited apoptosis accompanied by an increase in Bcl-2 level (P < 0.05), but the activity of Bax did not change (P > 0.05). AlCl(3) significantly reduced the viability of PC12 cells (P < 0.01). Curcumin increased cell viability in the presence of AlCl(3) (P < 0.01). The rate of apoptosis decreased significantly in the curcumin group (P < 0.05) when measured by flow cytometric analysis. Curcumin protected cells by increasing Bcl-2 level (P < 0.05), but the level of Bax did not change (P > 0.05).

CONCLUSIONSThis study demonstrates that curcumin improves the memory ability of AD mice and inhibits apoptosis in cultured PC12 cells induced by AlCl(3). Its mechanism may involve enhancing the level of Bcl-2.

Aluminum Compounds ; toxicity ; Alzheimer Disease ; drug therapy ; psychology ; Animals ; Apoptosis ; drug effects ; Cells, Cultured ; Chlorides ; toxicity ; Curcumin ; pharmacology ; therapeutic use ; Disease Models, Animal ; Female ; Learning ; drug effects ; Memory ; drug effects ; Mice ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Rats

OBJECTIVETo investigate the effect of nebulized total ginkgo flavone glycosides (TFG) on Th1/Th2 imbalance in mice with athma.

METHODForty-eight BALB/C mice were randomly divided into four groups: group A (control group, n=12); group B (asthmatic model group, n=12); group C (TFG nebulized treated group, n=12); group D (dexamethasone intraperitoneal treated group, n=12). The asthmatic model was established by sensitivity and local activation with Ovalbumin(OVA) and aluminum hydroxide Al(OH)3. TFG (50 g x L(-1), per aerosol per six mice, 30 minutes) was nebulized 20 days after modeling, while dexamethasone (1 g x L(-1)) was intraperitoneal once daily for 10 days. Perfusate of bronchoalveolar lavage fluid(BALF) was collected on day 32. The level of IL-4, IFN-gamma in BALF, and the level of total IgE in serum was determined. The airway inflammation pathology change and the expression of GATA-3 protein in lungs was detected by immunohistochemical staining.

RESULTCompared with model group, the decreased content of IL-4(49.30 +/- 7.95) ng x L(-1) and increased level of IFN-gamma (49.08 x 5.46) ng x L(-1). were found in BALF, and the level of total IgE (9.47 +/- 1.52) microg x L(-1) in serum also decreased in TFG treated group. In model group, smooth muscle hypertrophing, mucous hyperemia, mucous layer thickening, and inflammatory cell in filtration were observed. Phlegmasia was appeared in the bronchi, which was filled with lots of mucus. In contrast, the inflammatory reaction in TFG and Dexamethasone treated group was less obvious. Expression of GATA-3 was markly increased in model group with decreased expression in TFG treated group.

CONCLUSIONNebulized TFG showed a therapeutic effect for asthmatic mice, the mechanism may be explained by blockingnnnn GATA-3 expression and regulating the disorder Th1/Th2 imbalance.

Aluminum Hydroxide ; pharmacology ; Animals ; Asthma ; chemically induced ; drug therapy ; metabolism ; pathology ; Bronchoalveolar Lavage Fluid ; chemistry ; Dexamethasone ; therapeutic use ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Female ; Flavones ; Flavonoids ; chemistry ; GATA3 Transcription Factor ; metabolism ; Ginkgo biloba ; chemistry ; Glycosides ; chemistry ; therapeutic use ; Immunoglobulin E ; blood ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lung ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; pharmacology ; Random Allocation ; Th1 Cells ; drug effects ; metabolism ; Th2 Cells ; drug effects ; metabolism