OBJECTIVETo investigate the influence of microcystin-LR (MC-LR) on monocytes and lymphocytes in blood of mice and to find a sensitive index of toxic effects.

METHODSSpecific pathogen free Kunming male mice, aging 1 month-old,were randomly divided into 5 groups by weights, 7 mice for each group. The mice in 5 groups were exposed to MC-LR through intraperitoneal injection at 0, 3.125,6.250, 12.500 and 25.000 µg/kg respectively for 7 days. Then cytokine levels in the serum were measured by radioimmunoassay, DNA-protein crosslinks (DPC) was measured by the SDS/KCl precipitation technique, and the phagocytosis and ROS of leukocytes were detected by flow cytometry.

RESULTSThe levels of interleukin 6 in the 6.250, 12.500 and 25.000 µg·kg(-1)·d(-1) dose groups were (346.837 ± 25.536), (360.847 ± 37.886) and (434.245 ± 35.858)pg/ml respectively, which were significantly higher than those in the control group which the value was (232.775 ± 32.816) pg/ml (t values were -7.258, -6.760 and -10.966 respectively, P values were all < 0.05).While the level of tumor necrosis factor-alpha was(10.782 ± 0.966) fmol/ml in 25 µg·kg(-1)·d(-1) dose group was statistically lower than it in the control group which the value was (16.878 ± 3.378) fmol/ml (t value was 4.591, P < 0.05). The DPC levels of lymphocytes in 6.250, 12.500 µg·kg(-1)·d(-1) dose group were (242.576 ± 7.545),(241.472 ± 2.793) ng/ml,higher than it in the control group while the value was (228.657 ± 4.130) ng/ml (t value was -4.282, -6.801, P values were all <0.05). The fluorescence intensity of DCF in lymphocytes in the 4 treated groups were separately 3299.37 ± 120.54, 3281.38 ± 58.34, 3308.06 ± 136.12 and 3346.92 ± 108.69, all significantly lower than 3770.81 ± 131.39 in the control group (t values were 6.995, 9.007, 6.472 and 6.577 respectively, and P values were all <0.05). The fluorescence intensity of DCF in monocytes in the 4 treated groups (3271.51 ± 140.79, 3270.05 ± 117.92, 3326.90 ± 114.39 and 3292.49 ± 145.97 respectively) were also significantly lower than the value in the control group was 3841.72 ± 130.92 (t values were 7.847, 8.584, 7.835 and 7.411 respectively, P values were all <0.05). There was no significant difference in other index among the four experiment groups and the control group.

CONCLUSIONThe MC-LR administered via intraperitoneal injection to mice induced the alterations of some cytokines of monocytes and lymphocytes in blood. By comparison, the ROS of leukocyte was the most sensitive index.

Animals ; Cytokines ; metabolism ; Lymphocytes ; drug effects ; Male ; Mice ; Mice, Inbred Strains ; Microcystins ; pharmacology ; Monocytes ; drug effects ; Reactive Oxygen Species ; metabolism

OBJECTIVETo study how the way in which betaine promotes the proliferation of mouse spleen lymphocytes is related to calcium channels.

METHODBALB/c mice were used for this experiment. Mouse spleen lymphocytes were obtained through in vitro cultivation after they had been separated, and were divided into a negative control group, a Con A group, and 0.04, 0.4, 4, and 20 mmol x L(-1) betaine groups. MTT was used to observe the effect of betaine on the proliferation of mouse spleen lymphocytes; flow cytometry was used to measure the changes in the cell cycle of mouse spleen lymphocytes; and laser confocal scanning microscopy was used to observe the changes in the intracellular [Ca2+]i of mouse spleen lymphocytes after betaine or different calcium channel blockers were applied.

RESULTBetaine was found to promote the proliferation of mouse spleen lymphocytes 12 h after it had been applied in vitro in concentrations of 4 and 20 mmol x L(-1). It was also found to promote the proliferation of mouse spleen lymphocytes 24 h and 48 h after it had been applied in vitro in concentrations of 0.04, 0.4, 4, and 20 mmol x L(-1), with the effect being most marked for the 4 mmol x L(-1) group 24 h after its application. It was found to facilitate the entry of mouse spleen lymphocytes from the G0/G1 to the S phase 4, 6, 18, and 24 h after it had been applied to mouse spleen lymphocytes in a concentration of 4 mmol x L(-1), with the effect being most marked at 18 h after its application. Intracellular [Ca2+]i in mouse spleen lymphocytes increased significantly (P < 0.01) 6, 12, 18 h after 4 mmol x L(-1) betaine had acted on the lymphocytes, with the effect being most marked at 6 h. The calcium channel blockers nifidipine, diltiazem, mibefradil, and genistein had no effect on the increase of the intracellular [Ca2+]i in mouse spleen lymphocytes due to the application of betaine, while verapamil, mycifradin, heparin, and procaine could block such increase.

CONCLUSIONBetaine facilitates the entry of mouse spleen lymphocytes from the G0/G1 into the S phase by raising the intracellular [Ca2+]i in these cells, thus promoting their proliferation. Intracellular [Ca2+]i increases mainly in two ways: (1) By affecting the alpha1 subunit of the L-type voltage-gated calcium channel with mediation by G proteins and thus leading to an efflux of intracellular calcium: (2) By affecting the IP3R and RyR calcium channels of the intracellular calcium stores and thus leading to the release of intracellular calcium.

Animals ; Betaine ; pharmacology ; Calcium ; metabolism ; Calcium Channels ; metabolism ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Female ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C

OBJECTIVETo establish a new method for screening active ingredients of Chinese herbs by determining different bio-thermodynamic effects of 3 genosides on splenic lymphocyte of mice.

METHODSUsing a thermal bioactivity monitoring system, the maximum heat output (mHO), average metabolic heat (MH) and constant of decrease rate (DR) of lymphocyte were determined based on the growth metabolic power-time curve, and the outcomes were verified by MIT.

RESULTSThe mHO and MH increased and the DR decreased after lymphocytes being exposed to the 3 genosides in different concentrations, arranged upon their potency as genoside Rg3 > genoside Rg2 > genoside Rg1 (merely insignificant effect). MTT showed the same results.

CONCLUSIONHeat activity monitoring system could precisely display the different bio-thermal dynamic effects of 3 genosides on splenic lymphocyte.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Energy Metabolism ; drug effects ; Ginsenosides ; pharmacology ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; Thermodynamics

OBJECTIVETo observe the effect of Bawei Xilei powder on CD3, CD4, CD8 T-lymphocytes in peripheral blood and colonic mucosa of rat with ulcerative colitis.

METHODSixty SD rats were randomly divided into 6 groups, normal group, model group, low, middle and high dosage Bawei Xilei powder group, Sulfasalazine group. Ulcerative colitis was induced by immunization with rabbit 's colonic mucous emulsified with completely Freund's adjuvant in all rats. Rats in low, middle and high dosage Bawei Xilei powder group were administered with 0.05, 0.1, 0.2 mg Bawei Xilei powder for 18 days by enema respectively. While rats in Sulfasalazine group were enema administered with 100 mg Sulfasalazine, and the rats in other group were administered with equal volume of saline enema as control. We analyzed expression of CD3, CD4, CD8 T-lymphocytes in peripheral blood by flow cytometry and in colonic mucous by immunohistochemistry.

RESULTIn peripheral blood, compared with normal group, in model group, the increased of CD4 T-lymphocytes and CD4 /CD8 ratio, the reduced of CD8 T-lymphocytes, these results were significant discrepancy (P < 0.01). Compared with model group, after treatment with Bawei Xilei powder, CD8 T-lymphocytes increased, but only high dosage Bawei Xilei powder group had discrepancy (P < 0.05). But low dosage Bawei Xilei powder group, other treatment groups' rats showed CD4/CD8 ratio were reduced significantly (P < 0.05). In colonic mucous, compared with normal group, in model group, Rats showed that expression of CD3, CD4 T-lymphocytes reduced and CD8 T-lymphocytes increased obviously (P < 0.05, P < 0.01). Compared with model group, expression of CD8 T-lymphocytes reduced significantly in all treatment groups (P < 0.05, P < 0.01).

CONCLUSIONBawei Xilei powder may regulate their balance between T-lymphocytes subgroup, consequently relieve inflammatory injury in favor of ulcer reparation and tissue regeneration.

Animals ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; CD8 Antigens ; metabolism ; Colitis, Ulcerative ; immunology ; metabolism ; Colon ; drug effects ; metabolism ; pathology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Powders ; Rats ; T-Lymphocytes ; drug effects ; metabolism

This study was aimed to establish the quantitative analysis of hIL-2 in culture supernatant by multifunctional Luminex 100. The lymphocytes were separated from ACD-anticoagulated peripheral blood by density gradient method. The lymphocytes were stimulated with PHA for 48 hours, and frozen at -20 degrees C The relative fluorescence units of standard preparations and samples were detected by multifunctional Luminex 100, and the sample concentrations were calculated by standard curve. The results indicated that the regression equation of standard preparation is Lg (RFU) = 1.547 + 0.867 LgC. ANOVA F = 301.7427, p < 0.05 (nu = 6). The analysis of variance showed F = 301.7427, p < 0.05 (nu = 6). The test of regression coefficient showed t = 17.3707 (nu = 6), p < 0.05. It is concluded that method for induction and measurement of human IL-2 in vitro is established. The standard curve established by this way is statistically significant. There is linear relationship between the concentration of hIL-2 and fluorescence intensity.
Cell Separation ; methods ; Humans ; Interleukin-2 ; analysis ; Lymphocytes ; cytology ; drug effects ; metabolism ; Phytohemagglutinins ; pharmacology

OBJECTIVETo investigate the cytotoxicity of indium chloride (InCl₃) and its effects on micro-nucleus formation in primary human lymphocytes cultured in vitro.

METHODSThe CCK-8 assay was used to evaluate the cytotoxicity of 24 h exposure to different concentrations of InCl₃(4, 40, 80, 200, 500, and 1 000 µmol/L) in lymphocytes cultured in vitro. The cytokinesis-block method was used to determine the micronucleus level in lymphocytes exposed to different concentrations of InCl₃and the effects of anti-oxidant vitamin C on micronucleus frequency.

RESULTSLymphocytes exposed to InCl₃of no less than 500 µmol/L had significantly lower survival rates than those in the control group (P < 0.05). Lymphocytes exposed to 80 µmol/L InCl₃had a significantly higher micronucleus frequency than those in the control group (P < 0.05). However, there was no further increase in micronucleus frequency of lymphocytes exposed to 200 µmol/L InCl₃. Lymphocytes cultured in whole blood and exposed to 500 or 1000 µmol/L InCl₃had a significantly increased micronucleus frequency than those in the control group (P < 0.001). The increase in micronucleus frequency of lymphocytes induced by indium could be partially antagonized by 20 or 100 µmol/L vitamin C.

CONCLUSIONInCl₃can induce an increase in micronucleus frequency of primary human lymphocytes cultured in vitro, which might be associated with DNA damage induced by oxidative stress.

Cell Nucleus ; metabolism ; Cytokinesis ; DNA Damage ; Humans ; In Vitro Techniques ; Indium ; toxicity ; Lymphocytes ; drug effects ; Oxidative Stress

OBJECTIVETo study the effects of benzene and selenium on telomerase in mouse lymphocytes in vivo and evaluate telomerase activity as an early marker of benzene effects on lymphocytes.

METHODSMale Kunming mice were divided randomly into 8 groups, including negative control group, reagent control group, 100 mg/kg benzene group, 200 mg/kg benzene group, 400 mg/kg benzene group, 200 mg/kg benzene + 0.75 mg/kg selenium group, 200 mg/kg benzene + 1.50 mg/kg selenium group and 200 mg/kg benzene + 3.00 mg/kg selenium group, 5 mice in each group. The mice in different groups were treated with different methods, once daily for 5 days. After 48 hours of the final exposure, lymphocytes were separated and the telomerase activities were detected with TRAPELISA.

RESULTSCompared with negative and reagent control groups, the telomerase activity was increased after treatment with different dose of benzene and at the dose of 100 mg/kg benzene group it was significantly increased (P < 0.01). At the dose of 200 mg/kg benzene + 0.75 mg/kg selenium group, it was significantly increased (P < 0.01). Compared with the counterpart treated with 200 mg/kg benzene group, the expression of telomerase was increased at the different concentrations after treatment with benzene combined with selenium and it was significantly increased at the dose of 200 mg/kg benzene + 0.75 mg/kg selenium group (P < 0.05).

CONCLUSIONIncreased telomerase activity in lymphocytes stimulated by benzene at different concentrations indicates activation and proliferation of these lymphocytes of mice in vivo. Telomerase activity is probably a sensitive early marker of lymphocyte proliferation by benzene. Selenium can upregulate the telomerase activity.

Animals ; Benzene ; pharmacology ; Cells, Cultured ; Lymphocytes ; drug effects ; enzymology ; Male ; Mice ; Selenium ; pharmacology ; Telomerase ; metabolism

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Adult ; Apoptosis ; drug effects ; Benzene ; poisoning ; Biomarkers ; analysis ; Bone Marrow Cells ; drug effects ; Female ; Flow Cytometry ; Humans ; Leukocytes, Mononuclear ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Micronuclei, Chromosome-Defective ; drug effects ; metabolism ; Occupational Exposure ; analysis

OBJECTIVETo investigate the effect of spearmint oil on emphysema-like changes and the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta(IL-1beta), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-9) in lipopolysaccharide (LPS) treated rats.

METHODEmphysematous changes model was induced by intratracheal instillation of LPS once a week for up to 8 weeks in rats. Rats were divided into control, dexamethasone (0.3 mg x kg(-1)), and spearmint oil (10, 30,100 mg x kg(-1)) groups. Each group was treated with saline, dexamethasone, and spearmint of oil respectively for 4 weeks. Then total and different white blood cell counts in bronchoalveolar lavage fluid(BALF) were carried out. The pathologic changes of lung tissue such as alveolar structure, airway inflammation, and goblet cell metaplasia were observed by HE and AB-PAS staining. Expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9 were measured.

RESULTBoth spearmint and dexamethasone decreased the destruction of pulmonary alveolus. The total and different white blood cell counts in BALF including neutrophile and lymphocyte of spearmint oil 100 mg x kg(-1) and dexamethasone group were significantly reduced, and the goblet cell metaplasia was also inhibited. Dexamethasone had inhibitory effect on the expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9. Spearmint oil 30, 100 mg x kg(-1) significantly reduced TNF-alpha and IL-1beta respectively. Spearmint oil 10, 30 and 100 mg x kg(-1) had no effect on the expression of TIMP-1, but could decrease the expression of MMP-9 significantly in lung tissues.

CONCLUSIONSpearmint oil has protective effect on rats with emphysematous changes, since it improves alveolar destruction, pulmonary inflammation, and goblet cell metaplasia. The mechanism may include reducing TNF-alpha, IL-1beta content and inhibiting overexpression of matrix metalloproteinase-9 in lung tissues.

Animals ; Azo Compounds ; pharmacology ; Bronchoalveolar Lavage Fluid ; cytology ; Goblet Cells ; drug effects ; Interleukin-1beta ; drug effects ; metabolism ; Leukocytes ; drug effects ; metabolism ; Lipopolysaccharides ; Lymphocytes ; drug effects ; metabolism ; Matrix Metalloproteinase 9 ; drug effects ; metabolism ; Mentha spicata ; chemistry ; Metaplasia ; Monocytes ; drug effects ; metabolism ; Neutrophils ; drug effects ; metabolism ; Phytotherapy ; Plant Oils ; therapeutic use ; Pulmonary Emphysema ; chemically induced ; drug therapy ; enzymology ; pathology ; Rats ; Respiratory System ; drug effects ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; drug effects ; metabolism