Uniform design-comprehensive scoring method was used to investigate the effects of ethanol dosage, ethanol concentration and extraction time, based on the evaluation index from transfer rates of epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ, which are the main active components in Epimedii Folium. Furthermore, the optimum conditions for the ethanol extraction process were determined by multiple linear stepwise regression and empirical test. Then, the ethanol extract of Epimedii Folium prepared according to the optimized technological conditions was used to intervene the injury model of chondrocyte induced by interleukin-1 beta(IL-1β). Annexin V-FITC/PI staining flow cytometry was used to detect the apoptotic rate of chondrocyte and analyze the effect of ethanol extract of Epimedii Folium on chondrocyte injury model. The optimum conditions of ethanol extraction were as follows. Crude powder of Epimedii Folium was added with 18 times of 70% ethanol solution, and extracted for twice in the refluxing process, for 60 minutes each time. Under the conditions, the extraction rates of the above five active components were 94.21%, 94.76%, 93.85%, 96.17% and 96.85%, respectively. The optimized ethanol extraction process of Epimedii Folium was reasonable, feasible and reproducible. This ethanol extract could significantly reduce the early apoptotic rate, late apoptotic and necrotic rate, total apoptotic rate(P<0.05 or P<0.01) of chondrocyte injury model induced by IL-1β, suggesting that the ethanol extract of Epimedii Folium can inhibit the apoptosis of chondrocytes induced by IL-1β to a certain extent, which lays a foundation for further study on its prevention and treatment of osteoarthritis.
Chondrocytes/drug effects* ; Epimedium/chemistry* ; Ethanol ; Humans ; Interleukin-1beta ; Plant Extracts/pharmacology* ; Plant Leaves/chemistry*

BACKGROUNDInterleukin (IL)-1beta may effectively decrease introcular pressure (IOP) when administered by subconjunctival injection in normal rabbit. However, IL-1beta is a large molecular agent and an inflammation factor. The aim of this study was to evaluate the penetrability of IL-1beta, and the concentrations of both tumor necrosis factor (TNF)-alpha and IL-6 in the aqueous humor of normal rabbits treated with IL-1beta.

METHODSA total of 170 rabbits were used in the study and were assigned to several different treatment groups as follows: 125 of the rabbits were assigned to two groups. In one group, 33 rabbits were injected subconjunctivally with IL-1beta and 39 were injected with saline alone. In the other group, 27 rabbits were given eye drops containing IL-1beta (400 ng/ml) and 26 were given saline alone. Aqueous humor (AH) was drawn and the concentration of IL-1beta within the fluid measured. The IOP was measured in another six rabbits after administration of eye drops containing IL-1beta (400 ng/ml). A further 20 rabbits were assigned to 3 groups as follows: eight untreated normal controls; six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of TNF-alpha in the fluid was measured. Another 19 rabbits were assigned to 3 groups as follows: seven untreated normal controls; and six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of IL-6 in the fluid measured. Measurement of cytokine concentration was by radio-immunoassay in all cases.

RESULTSThe IL-1beta concentration in the AH was higher in those animals in which it had been administered subconjunctivally (P < 0.01). The IL-1beta concentration in the AH of the animals given eye drops was almost the same as that in the controls (P > 0.05). The administration of IL-1beta in the form of eye drops had little effect upon IOP reduction. Lower TNF-alpha concentrations were seen in the AH after the subconjunctival administration of IL-1beta, but the concentration of IL-6 was the same as in the normal controls.

CONCLUSIONSIL-1beta shows good corneal penetrability after subconjunctival injection into normal rabbit eyes. The IOP reduction induced by IL-1beta is unlikely be associated with an inflammatory response.

Animals ; Aqueous Humor ; drug effects ; metabolism ; Interleukin-1beta ; pharmacology ; Interleukin-6 ; metabolism ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect of one of the acute-phase proteins, fibrinogen, on the release of IL-1beta and -8 by human peripheral polymorphonuclear leukocytes (PMN) and the possible role of fibrinogen during the destruction of periodontium.

METHODSPeripheral PMN were isolated by discontinuous density gradient centrifuging technique. The freshly isolated PMN were suspended in Hank's balanced saline solution (1 x 10(9)/L) supplemented with 0.5% BSA and 0.1% glucose. The levels of IL-1beta and -8 in the supernatants produced by cultured cells upon the addition of human fibrinogen at different concentrations were measured by ELISA technique.

RESULTSIncubated with human fibrinogen at 2 g/L or 10 g/L for different time periods, human peripheral PMN released significantly greater amount of IL-1beta [(10.41 +/- 0.37) - (35.86 +/- 0.30) ng/L or (22.81 +/- 0.45) - (57.77 +/- 2.08) ng/L] and IL-8 [(93.90 +/- 13.95) - (2045.66 +/- 53.03) ng/L or (115.02 +/- 10.61) - (3858.69 +/- 25.65) ng/L] than PMN without the stimulation of fibrinogen (IL-1beta, P < 0.001, and IL-8, P < or = 0.016). The higher concentration of fibrinogen or the longer treatment time, the higher levels of IL-1beta and -8 were released by PMN (P < 0.001).

CONCLUSIONSFibrinogen induced the secretion of pro-inflammatory cytokines IL-1beta and -8 by PMN and may be involved in magnification of the inflammatory response of periodontium and bone resorption.

Cells, Cultured ; Fibrinogen ; pharmacology ; Humans ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Middle Aged ; Neutrophils ; drug effects ; secretion

This work was aimed to investigate the effect of quinacrine on peripheral granulocytes and lymphocytes, interleukin 1 (IL-1) and interleukin 6 (IL-6) in peripheral blood serum of inflammatory reaction induced by microwave irradiation, and observe the protective effect of quinacrine against microwave irradiation injury. BALB/c mice were suffered from microwave irradiation with the total intensity of 50 mW/cm(2) for 30 minutes, at 1 hour before irradiation quinacrine (12.6 mg/kg or 50.4 mg/kg) was orally administrated. Mice received same volume of water for injection instead of quinacrine were named as microwave irradiation group (MR group), and mice received no microwave irradiation but stayed in microwave irradiation environment also for 30 min were set as control. After microwave irradiation, mice were sacrificed and peripheral blood cells were analyzed with cytoanalyzer, and mice serum interleukin-1β, interleukin-6 were detected by radioimmunoassay. The results showed that microwave irradiation increased the count of peripheral granulocytes and lymphocyte along with prolongation of time, while the increase of these cells in mice administrated quinacrine was markedly delayed. The level of IL-1β in serum of mice was significantly increased after 1 day of microwave irradiation (50 mW/cm(2)), and recovered to normal level after 7 days. The 2 concentrations of quinacrine (12.6 mg/kg, 50.4 mg/kg) could suppress level of IL-1β in serum induced by microwave irradiation. The level of IL-6 in serum of mice was gradually increased after microwave irradiation with intensity of 50 mW/cm(2) for 7 days, but quinacrine administration could delay the rise of IL-6 level, specially within time of 2 days. It is concluded that the quinacrine administration can delay the increase of peripheral granulocytes and lymphocytes inducted by microwave irradiation, and may partially suppress the rise of IL-1β and IL-6 in serum. The results of this study suggest that the quinacrine can provide some protective effect against microwave irradiation injury.
Animals ; Inflammation ; Interleukin-1 ; blood ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Leukocyte Count ; Male ; Mice ; Mice, Inbred BALB C ; Microwaves ; adverse effects ; Quinacrine ; pharmacology

OBJECTIVETo test the effect of high-mobility group box protein 1 (HMGB1) alone or in synergy with interleukin-1β (IL-1β) on the expression of IL-8 in human airway epithelial cells in vitro.

METHODSHuman airway epithelial 16HBE and A549 cell lines were incubated with HMGB1 (100 ng/ml) in the absence or presence of IL-1β (10 ng/ml) for 24 h, and the changes of IL-8 mRNA and protein expressions were assessed using quantitative PCR and enzyme-linked immunosorbent assay (ELISA).

RESULTSIn the two human airway epithelial cell lines, HMGB1 alone did not produce obvious effect on the expression of IL-8, but in the presence of IL-1β, HMGB1 caused a significant increase of IL-8 expressions at both the mRNA and protein levels.

CONCLUSIONHMGB1 in synergy with IL-1β increases the expression of IL-8 in human airway epithelial cells, which provides new evidence that HMGB1 contributes to neutrophilic airway inflammation by regulating IL-8 expression.

Bronchi ; cytology ; drug effects ; Cell Line ; Epithelial Cells ; drug effects ; metabolism ; HMGB1 Protein ; pharmacology ; Humans ; Inflammation ; Interleukin-1beta ; pharmacology ; Interleukin-8 ; metabolism ; RNA, Messenger

OBJECTIVETo explore and identify the method for IL-1beta induced New Zealand rabbit knee chondrocyte degeneration, thus providing experimental bases for Chinese medical research on osteoarthritis from in vitro cultured chondrocytes.

METHODSUnder aseptic conditions, bilateral knee joint cartilage was collected from 4-week old New Zealand rabbits. Chondrocytes were separated by type II collagenase digestion and mechanical blowing method. They were randomly divided into two groups when passaged to the 2nd generation, the normal control group (group Z) and the IL-1beta induced model group (group M). No intervention was given to those in group Z. 10% FBS culture media containing 10 ng/mL IL-1beta was added to group M. All cells were passaged to the 3rd generation. They were compared using morphological observation, toluidine blue staining, type II collagen immunohistochemical staining, and flow cytometry.

RESULTSUnder inverted microscope, the second and the 3rd generation chondrocytes' phenotype of group Z was stable with good proliferation. Most cells turned into fusiform and slabstone shaped. In group M, most cells turned into long spindle shape or irregular shape. Results of toluidine blue staining and immunohistochemistry showed that the positive expression of chondrocytes after staining in group Z was superior to that in group M. Results of flow cytometry showed that there was statistical difference in the apoptosis rate of the second generation chondrocytes between group M and group Z (P < 0.01).

CONCLUSIONIt was obviously seen that chondrocytes in IL-1beta induced New Zealand rabbit knee chondrocyte model obviously degenerated, which could be used in related experimental researches on osteoarthritis.

Animals ; Cartilage ; cytology ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; Interleukin-1beta ; pharmacology ; Knee Joint ; cytology ; drug effects ; Rabbits

OBJECTIVETo study the effect of interleukin-1β (IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis.

METHODSCultured HESCs were stimulated with 250, 500, and 750pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay.

RESULTSIL-1β treatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs.

CONCLUSIONIL-1β can affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.

Activins ; metabolism ; Cells, Cultured ; Endometriosis ; metabolism ; Endometrium ; cytology ; Female ; Humans ; Interleukin-1beta ; pharmacology ; Stromal Cells ; drug effects ; metabolism

This study compared the biological changes of lipopolysaccharide (LPS)-treated dental pulp (DP) cells directly cultured on mineral trioxide aggregate (MTA) and calcium silicate (CS) cements. DP cells were treated with LPS for 24 h. Then, the LPS-treated DP cells were cultured on MTA or CS cements. Cell viability, cell death mechanism and interleukin (IL)-1β expressions were analysed. A one-way analysis of variance was used to evaluate the significance of the differences between the means. A significantly higher IL-1β expression (2.9-fold) was found for LPS-treated cells (P<0.05) compared with DP cells without LPS treatment at 24 h. Absorbance values of LPS-treated cells cultured on CS cement were higher than a tissue culture plate. A significant difference (P<0.05) in cell viability was observed between cells on CS and MTA cements 24 h after seeding. At 48 h, a high concentration of Si (5 mM) was released from MTA, which induced LPS-treated DP cell apoptosis. The present study demonstrates that CS cement is biocompatible with cultured LPS-treated DP cells. MTA stimulates inflammation in LPS-treated DP cells, which leads to greater IL-1β expression and apoptosis.
Calcium Compounds ; Dental Cements ; Dental Pulp ; drug effects ; metabolism ; Humans ; Inflammation ; chemically induced ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; pharmacology ; Silicates

OBJECTIVETo investigate the inhibitory effect of resveratrol on interleukin-1β (IL-1β) production in mesenchymal stem cells (MSCs) exposed to radiation and the action mechanism of resveratrol.

METHODSMSCs were divided into blank control group, radiation group, shRNA interference group, and resveratrol groups. The resveratrol groups were given different doses of resveratrol (50, 100, and 200 µmol/L) before radiation. The secretion and expression of IL-1β was measured by enzyme-linked immunosorbent assay, Western blot, and RT-PCR.

RESULTSCompared with the radiation group, the resveratrol groups had significantly decreased extracellular secretion of IL-1β (t = 83.34, 24.48, and 12.52, P < 0.05 for all) and significantly decreased intracellular expression of IL-1β protein and mRNA (t = 8.695, 14.77, and 13.9, P < 0.05 for all). Compared with those given 200 µmol/L resveratrol alone before radiation, the MSCs treated by SIRT1 silencing and given 200 µmol/L resveratrol before radiation had significantly increased extracellular secretion of IL-1β (t = 18.57, P < 0.05) and significantly increased intracellular expression of IL-1β protein and mRNA (t = 10.24, P < 0.05).

CONCLUSIONResveratrol can significantly inhibit the production of IL-1β in MSCs exposed to radiation, and SIRT1 may play a key regulatory role in the process of inflammation induced by radiation.

Cells, Cultured ; Humans ; Interleukin-1beta ; metabolism ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; radiation effects ; Radiation ; Radiation Dosage ; Stilbenes ; pharmacology

OBJECTIVE: To detect cyclooxygenase-2 (COX-2) expression and prostaglandin E2 (PGE2) release in human nasal epithelia (HNE) induced by hypoxia and/or IL-1beta of different time gradient, and to investigate their roles in nasal inflammatory pathogenesis. METHOD: Western Blot and fluorescent real time quantitative PCR were performed to detect the expression of COX-2 in HNE induced by hypoxia and/or IL-1beta. The concentrations of PGE2 were determined by enzyme immunoassay. Median comparison was statistically treated by rank sum test, and generalized linear model was used to analyze the association of hypoxia with IL-1beta. RESULT: Weak expressions of COX-2 and PGE2 were detected in normal HNE. COX-2 expression and PGE2 release increased in HNE induced by hypoxia and/or IL-1beta in time-dependent manner. Stronger expressions of COX-2 and PGE2 induced by hypoxia and/or IL-1beta than control were detected on different time (P < 0.05). The strongest inducible effect was found in hypoxia+IL-1beta group, and inducible effect decreased in hypoxia group and IL-1beta group in turn. The expressions of COX-2 and PGE2 in hypoxia+IL-1beta group were more than the sum of hypoxia group and IL-1beta group on same time. CONCLUSION Hypoxia and/or IL-1beta effectively induce COX-2 expression and PGE2 release in HNE. Synergistic effect between hypoxia and IL-1beta has been found in induction of COX-2 and PGE2 in HNE. Results indicate that the increased expressions of COX-2 and PGE2 are involved in inflammation of HNE induced by hypoxia and/or IL-1beta in vitro.
Cell Hypoxia ; Cells, Cultured ; Cyclooxygenase 2 ; metabolism ; Dinoprostone ; metabolism ; Humans ; Interleukin-1beta ; pharmacology ; Nasal Mucosa ; cytology ; metabolism