Immunogenicity for medical devices of animal origin is the key and difficult point during immune safety evaluation for these devices. This paper firstly investigated the effect of collagen sponge of animal origin on mouse splenic lymphocyte transformation and proliferation, and then analyzed the influence factors on the MTT method and CFSE method. The results showed that collagen sponge extract cannot significantly induce transformation and proliferation of mouse splenic lymphocyte in vitro.
Animals ; Cells, Cultured ; Collagen ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Porifera ; chemistry ; Spleen ; cytology

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 investigate the DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide (CS2) in the implantation phase and to explore the mechanism of abnormal implantation induced by CS2 from the perspective of immune injury.

METHODSMice were exposed to CS2 at different doses or at different time points in the implantation phase to establish model 1 and model 2. For model 1, mice were assigned to four groups to receive a single intraperitoneal injection of low-dose CS2 (0.1 LD50, 157.8 mg/kg), middle-dose CS2 (0.2 LD50, 315.7 mg/kg), and high-dose CS2 (0.4 LD50, 631.4 mg/kg) as well as an equal volume of olive oil (control) on gestational day (GD) 4. For model 2, mice were assigned to four groups to receive a single intraperitoneal injection of CS2 (0.4 LD50, 631.4 mg/kg) or an equal volume of olive oil (control) on GD3, GD4, GD5, and GD6. At the end, single cell suspension of splenic lymphocytes was prepared. Cell viability was measured by trypan blue staining, and the DNA damage of splenic lymphocytes was evaluated by alkaline single cell gel electrophoresis assay.

RESULTSThe middle-dose and high-dose exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P < 0.01); there was significant regression relationship between indicators of DNA damage and exposure doses (P < 0.01). The GD3, GD4, GD5, and GD 6 exposure groups showed significantly more DNA damage of splenic lymphocytes than the control group (P < 0.01), and the GD 4 exposure group had the most DNA damage.

CONCLUSIONExposure to CS2 in the implantation phase can induce DNA damage of splenic lymphocytes in pregnant mice, and the DNA damage was aggravated with the increase in CS2 concentration. GD4 may be the sensitive time point for DNA damage of splenic lymphocytes induced by CS2 in pregnant mice.

Animals ; Carbon Disulfide ; toxicity ; DNA Damage ; drug effects ; Embryo Implantation ; Female ; Lymphocytes ; drug effects ; Mice ; Pregnancy ; Spleen ; cytology

The purpose of this paper is to study the effect of kinetin (Kn) on immunity and splenic lymphocyte proliferation in vitro of aging rats induced by D-galactose (D-gal). Fifty SD rats were randomly divided into five groups: control group, aging model group, Kn low dose group, Kn middle dose group and Kn high dose group. The aging model group was proposed by napes subcutaneous injection of D-gal (125 mg/kg) for 45 d, and anti-aging groups were intragastrically administered with 5, 10, 20 mg/kg of Kn respectively from day 11. IgG, IgA, IgM contents of serum, the apoptosis percentage, stimulation index (SI) and proliferation index (PI) of splenic lymphocyte in vitro were evaluated. The results showed that the apoptosis percentage of splenic lymphocyte in aging model rats was higher, the serum IgG, IgA and IgM contents, SI and PI were lower than control group. Kn significantly decreased the apoptosis percentage of splenic lymphocyte, while increased the serum IgG, IgA and IgM contents, SI and PI in aging model group. These results suggest that Kn could inhibit the apoptosis, while promote the proliferation of splenic lymphocyte, and then effectively enhance the immune power of the aging rats and slow down the aging process.
Aging ; drug effects ; immunology ; Animals ; Antibodies ; blood ; Apoptosis ; Cell Proliferation ; drug effects ; Galactose ; adverse effects ; Kinetin ; pharmacology ; Lymphocytes ; cytology ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology

OBJECTIVETo study the influences of different processing methods on the content of Angelica sinensis polysaccharides (APS) from the same origin.

METHODThe contents of neutral polysaccharides and acidic polysaccharides in various samples of A. sinensis were determined by phenol-sulfuric acid and carbazole-sulfuric acid method, respectively. The proliferation ability of lymphocyte was detected by MTT method after the cells were cultured with different concentrations of APS from two samples processed by different methods.

RESULTThe different processing methods had different effects on the contents of polysaccharide. The maximum content of APS (26.03%) was found in the sample processed by microwave drying medium-fired, but the minimum content of APS (2.25%) was found in the sample processed by vacuum drying at 50 TC. Furthermore, the APS (high concentration group, P < 0.01) processed by microwave drying medium-fired could both accelerate proliferation of spleen lymphocytes directly and increase proliferation of T cells of mice induced by Con A. However, the APS processed by far-infrared drying did not show conspicuous immune enhancement activity.

CONCLUSIONDifferent processing methods have different effects on the contents of APS and the proliferation ability of lymphocytes.

Angelica sinensis ; chemistry ; Animals ; Cell Proliferation ; drug effects ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Mice ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology ; Spleen ; cytology ; T-Lymphocytes ; cytology ; drug effects

OBJECTIVETo study the effect of supplemented Danggui Shaoyao San (DSS) and its disassembled prescriptions on function of lymphocyte in aged rats.

METHODSRats aged 18 months were administered with decoction of DSS and its disassembled prescriptions separately in groups for 3 weeks to prepare drug containing serum. Samples of ConA induced rats splenic lymphocytes proliferation was treated respectively with prepared drug serum, for testing their effect on lymphocyte proliferation using methyl thiazolyl tetrazolium (MTT) method, and isoprel (ISO) and propranolol were taken as the controls.

RESULTSThe A value of lymphocyte proliferation in the drug serum treated groups was significantly different from that in the control groups (P < 0.01), which could be reduced markedly after treatment with ISO, but could restore to the level before ISO treatment by adding drug serum or propranolol.

CONCLUSIONDSS and/or its disassembled prescriptions could raise the lymphocyte proliferation in aged rats significantly, it also shows antagonizing effect against the inhibition of ISO on lymphocyte proliferation.

Aging ; Animals ; Cell Division ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; T-Lymphocytes ; cytology ; drug effects

AIMTo observe the effects of acute hypoxia and adenosine on splenic T lymphocyte proliferation.

METHODSWistar rats were divided into control and hypoxic group, and the latter were exposed to hypoxia (5000 m simulated high altitude, 23 h/d). Three days later, spleen cells were collected and stimulated by 5.0 microg/ml and 2.5 microg/ml concanavalin A (ConA) to determine the splenocyte proliferation. The proliferation was also observed after addition of different amount of adenosine to culture medium.

RESULTSAcute hypoxia and adenosine had marked inhibitory effect on mitogenic response to Con A in splenic T cells, and the inhibitory effect induced by adenosine displayed concentration-dependent.

CONCLUSIONAcute hypoxia may impair the T cell function and adenosine could be involved in this process.

Adenosine ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Concanavalin A ; pharmacology ; Culture Media ; chemistry ; Hypoxia ; Male ; Rats ; Rats, Wistar ; Spleen ; cytology ; T-Lymphocytes ; cytology ; drug effects

A water-soluble polysaccharide fraction from root of Potentilla anserine was obtained. Gas chromatogram, FT-IR, physical and chemical characteristics of the Potentilla anserine polysaccharide fraction (PAPF) were analyzed. The protective effects of PAPF against the H2O2 induced process of apoptosis of murine splenic lymphocytes were investigated in vitro. Morphological assessment of apoptosis was performed with light microscope and laser scanning confocal microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. The results showed that PAPF is composed of rhamnose, arabinose glucose and galactose. H2O2 (200 micromol x L(-1)) induced apoptosis of murine splenic lymphocytes with the cell volume reduced, cytoplasm and nuclear shrunk and DNA stained non-uniformly. Condensed chromatin and formation of apoptotic body were observed in the apoptotic cells. Apoptotic bodies in the cells treated with PAPF and H2O2 were less than those in H2O2 treatment alone. DNA fragmentation assay showed that PAPF (50, 100, 200, and 400 microg x mL(-1)) obviously reduced H2O2-induced ladder bands. Flow cytometry analysis showed that H2O2 increased the populations of apoptotic sub-G1 cells from 5.60% (control) to 45.40%, and PAPF decreased H2O2-induced apoptosis to 37.80%, 22.70%, 17.70%, and 8.50%, respectively. In conclusion, PAPF reduced H2O2-induced oxidative damage in a dose dependent manner.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; DNA Fragmentation ; Flow Cytometry ; Hydrogen Peroxide ; pharmacology ; Lymphocyte Count ; Lymphocytes ; cytology ; drug effects ; Mice ; Polysaccharides ; pharmacology ; Potentilla ; Spleen ; cytology

The objective of this study was to explore the immunomodulatory effects of betulinic acid (BA) extracted from the bark of white birch on mice. Female mice were orally administered BA for 14 days in doses of 0, 0.25, 0.5, and 1 mg/kg body weight. We found that BA significantly enhanced the thymus and spleen indices, and stimulated lymphocyte proliferation induced by Concanavalin A and lipopolysaccharide as shown by MTT assay. Flow cytometry revealed that BA increased the percentage of CD4+ cells in thymus as well as the percentage of CD19+ and the ratios of CD4+/CD8+ in spleen. BA increased the number of plaque-forming cell and macrophage phagocytic activity as indicated by a neutral red dye uptake assay, and the peritoneal macrophages levels of TNF-alpha were also increased. In contrast, serum levels of IgG and IgM and serum concentrations of IL-2 and IL-6 were significantly decreased in BA-treated mice compared to the control as assayed by haemagglutination tests and ELISA, respectively. Taken together, these results suggest that BA enhances mouse cellular immunity, humoral immunity, and activity of macrophages. Thus, BA is a potential immune stimulator and may strengthen the immune response of its host.
Adaptive Immunity/*drug effects ; Animals ; Betula/*chemistry ; Cell Proliferation/drug effects ; Cytokines/blood ; Female ; Immunity, Innate/*drug effects ; Immunologic Factors/*pharmacology ; Macrophages/drug effects ; Mice ; Phagocytosis/drug effects ; Random Allocation ; Spleen/cytology ; Thymus Gland/cytology ; Triterpenes/*pharmacology

AIMTo study effect of soybean isoflavones (SI) on spleen in radiated mice.

METHODS90 male mice were randomly divided into control group, radiated group, radiated plus 0.5% dose SI group. After 2-week feeding, the mice received 4.0 Gy 137Cs gamma-radiation, the cell cycles, cell apoptosis and proliferation on the spleen and the spleen index were observed in radiated after 12 h, 24 h, 1 week and 2 weeks.

RESULTSAfter the mice were radiated, the spleen were significantly atrophy, the rate of the cell apoptosis and the cell cycles of G0-G1 phase in splenocytes were significantly increased (P < 0.01), the cell cycles rate of S phase and the proliferation index were significantly decreased in spleen (P < 0.05). Compared with radiated group, the spleen atrophy and the rate of the cell cycles of G0-G1 phase were significantly decreased (P < 0.05), and the cell cycles of G2-M phase and the proliferation index were significantly increased (P < 0.05) in the mice supplied 0.5% soybean isoflavones.

CONCLUSIONThe soybean isoflavones could significantly increase spleen radioprotective effect in mice.

Animals ; Apoptosis ; drug effects ; radiation effects ; Cell Cycle ; drug effects ; radiation effects ; Cellular Structures ; Isoflavones ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Radiation, Ionizing ; Soybeans ; Spleen ; cytology