OBJECTIVETo compare the effect of three preparations of xiaoyao formula (xiaoyao capsule, xiaoyao pill and Xiaoyao decoction) on soothing liver and strengthening spleen.

METHODThe liver depression and spleen deficiency rat model were established by forcing swimming, confining movement test and feeding on alternate days. The rats were randomly divided into eight groups, namely the blank group, the model group, the Cisapride group (2.7 mg x kg(-1)), the xiaoyao decoction group (1.62 g x kg(-1)), the xiaoyao pill group (1.62 g x kg(-1)) and the Xiaoyao capsule groups of high dose (3.24 g x kg(-1)), medium dose (1.62 g x kg(-1)) and low dose (0.81 g x kg(-1)), with intragastric administration for 3 weeks. The contents of NE, DA, 5-HT in serum were measured by HPLC-electrochemical detection method. The contents of motilin (MTL) and somatostation (SS) in plasma was determined by radioimmunassay. The expressions of MTL and SS in tissue were determined by immunohistochemical method.

RESULTThe contents of NE and MTL in the xiaoyao decoction group was significantly higher than that in the model group, while the content of 5-HT and SS was significantly lower than that in the model group (P < 0.01). The expression of MTL in the xiaoyao capsule group (1.62 g x kg(-1)) was significantly higher than that in the model group and the contents of 5-HT and SS was significantly lower than that in the model group (P < 0.05). The mean optical density of MTL in gastrointestinal tissue of rats in the xiaoyao decoction group and the xiaoyao capsule group (1.62 g x kg(-1)) was remarkably higher than that in the model group, while the expression of SS was notably lower than that in the model group (P < 0.05).

CONCLUSIONAll of the three preparations of Xiaoyao formula have the effect on soothing liver and strengthening spleen. Xiaoyao decoction shows better effect than xiaoyao capsule with same dosage, while xiaoyao capsule shows better effect than xiaoyao pill with same dosage.

Animals ; Body Weight ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Intestines ; drug effects ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Motilin ; metabolism ; Rats ; Somatostatin ; metabolism ; Spleen ; drug effects ; metabolism ; Stomach ; drug effects ; metabolism

To study the potential effect of Dioscorea nipponica(DN) in intervening peripheral system of rats based on metabolomic analysis. The identification of the potential intervention targets of DN in peripheral system may facilitate its safe application and therapeutic potential exploitation. Totally 20 male SD rats were randomly divided into the blank group and the DN-treated groups, with 10 rates in each group. The DN-treated group was orally administrated with DN extracts once a day for 5 days, with the dose of 80 mg x kg(-1) (equivalent to 15 g crude drug in human), and the blank group was given equal volume of saline once a day for 5 days. Heart, liver, spleen, lung, and kidney tissues and serum samples were collected from each rat 24 h later after the last administration. The ultra-performance liquid chromatography/quadrupole time-of-flight-mass spectrometry based metabolomics was used to investigate the effect of DN in intervening peripheral system of rats. After the treatment with DN, 5 modulated metabolites in heart tissue, 6 in liver tissue, 5 in spleen tissue, 3 in lung tissue, 5 in kidney tissue and 6 in serum sample were identified and considered as the potential intervention targets of DN. Effect of DN in regulating some endogenous metabolites was beneficial for protecting peripheral system, while that in other endogenous metabolites produced potential toxicity to peripheral system. The metabolomic analysis revealed the coexistence of protective and toxic effects of DN on peripheral system, which may be a practical guidance for its safe application and beneficial to the expansion of its application scope.
Animals ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Heart ; drug effects ; Kidney ; chemistry ; drug effects ; metabolism ; Liver ; chemistry ; drug effects ; metabolism ; Lung ; chemistry ; drug effects ; metabolism ; Male ; Metabolomics ; Rats ; Rats, Sprague-Dawley ; Spleen ; drug effects ; metabolism

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

OBJECTIVE: To explore the effect of dexmedetomidine (DEX) on high-mobility group box 1 protein (HMGB1) in the serum and spleen in rats with sepsis and the underlying mechanisms.
 METHODS: A total of 100 male Wistar rats were randomly divided into five groups: a control group, a sepsis group, a DEX group, an α-bungarotoxin- tetramethylrhodamine (α-BGT) plus DEX group and an α-BGT group (n=20 in each group). In the control group, same volume of normal saline (NS) was administrated, while lipopolysaccharide (LPS) was injected to the vein to develop a classic sepsis model. At the 48th h after LPS injection, the rats were sacrificed, and the blood and spleen were collected. The content of tumor necrosis factor-α (TNF-α) and HMGB1 in blood serum were detected by enzyme-linked immuno sorbent assay (ELISA), and the HMGB1 in the spleen was examined by Western blot.
 RESULTS: Among the 5 groups, there were statistical significance in the mortality, the serum level of TNF-α and HMGB1, and the content of secreting type of HMGB1 in spleen (all P<0.05), and the serum level of HMGB1 was positively correlated with the content of secreting type of HMGB1 (r=0.863, P<0.05).
 CONCLUSION DEX exert late anti-inflammatory effects in the serum and spleen in rats with sepsis, which is related to the activation of the cholinergic anti-inflammatory pathway.
Animals ; Dexmedetomidine ; pharmacology ; HMGB1 Protein ; blood ; metabolism ; Lipopolysaccharides ; Male ; Rats ; Rats, Wistar ; Sepsis ; drug therapy ; metabolism ; Spleen ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; blood

Iron deficiency anemia is one of the most common micronutrient deficient conditions around the globe with various consequences, including the weakened immune system. Quercetin is widely distributed bioflavonoid; it has been debated for its dual roles in iron regulation. Quercetin-iron interaction in the body is a complex mechanism which has not been completely understood. The present study aimed to investigate the effect of quercetin on iron supplementation in iron deficiency anemia and on iNOS expression in splenic macrophages. The rat model of iron deficiency anemia was induced by feeding low iron diet to weanling rats for 20 days. The animals were then administered with ferrous sulfate, quercetin, and their combination for 30 days. Blood parameters, histopathological analysis, iron storage, CD68, iNOS and SLC40 expression in rat spleen were investigated. Our results showed that quercetin regulated iron absorption, despite SLC40 down-expression, indicating possible alternate route of iron transport, and that quercetin modulated iNOS production in splenic macrophages.
Anemia, Iron-Deficiency ; drug therapy ; genetics ; metabolism ; Animals ; Dietary Supplements ; analysis ; Female ; Homeostasis ; drug effects ; Humans ; Iron ; deficiency ; Macrophages ; drug effects ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Quercetin ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Spleen ; drug effects ; enzymology

This study was aimed to explore the effect of cordyceps sinensis enhancing lymphocyte proliferation and surface CD marker expression in simulated microgravity environment. The splenic lymphocytes were separated from mice and cultured in the rotary cell culture system simulated microgravity environment. The cells were treated with different concentration of cordyceps sinensis solution (0, 6.25, 12.5, 25 and 50 µg/ml) for 24, 48 and 72 h respectively, then the cells were harvested, and analyzed for cell proliferation and the expression of cell surface markers (CD4 and CD8). The results showed that under simulated microgravity environment, the lymphocyte proliferation was inhibited. When the concentration of cordyceps sinensis was 25 or 50 µg/ml, the lymphocyte proliferation, CD4 and CD8 expressions all increased, but 50 µg/ml cordyceps sinensis could inhibit the proliferation ability with the time prolonging. It is concluded that the suitable concentration of cordyceps sinensis displayed the ability to enhance the lymphocyte proliferation and CD marker expression in simulated microgravity environment. These results may be valuable for screening drugs which can be potentially against immunosuppression under simulated microgravity.
Animals ; CD4 Antigens ; metabolism ; CD8 Antigens ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cordyceps ; Immune Tolerance ; Lymphocyte Activation ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Polysaccharides ; pharmacology ; Spleen ; cytology ; Weightlessness Simulation

BACKGROUNDEpidermal burn injury may trigger significant apoptosis of the spleen cells, which might be caused by a burn-induced systemic inflammatory reaction. Heparin has been shown to possess anti-inflammatory properties. Interleukin 1 (IL-1) is centrally important among pro-inflammatory cytokines. We hypothesized that heparin might inhibit burn-induced apoptosis in the spleen via suppression of the IL-1 pathway.

METHODSBurn injury was performed on IL-1 R+/+ ( IL-1 receptor wild-type mouse) and IL-1 R-/- (IL-1 receptor knock-out mouse) mice, and they were then treated with heparin, saline or IL-1 receptor antagonist IL-Ra. Apoptosis, IL-1α and IL-1β expression were assessed in the spleens and serum. Survival curve analysis was further applied to elucidate the mechanism of heparin's protective properties.

RESULTSBurn induced significant apoptosis (sham: 3.6%± 2.1% vs. burn: 28.8%± 5.9%; P < 0.001) and remarkable expression o IL-1α and IL-1β in the mouse spleens and serum. Heparin reduced the burn-induced apoptosis in the spleens (heparin treated: 8.6%± 3.4%, P < 0.005), which could be blocked by IL-1Ra. Heparin markedly decreased both IL-1α and IL-1β expression in the spleens and serum of burned mice. IL-1 R-/- mice demonstrated considerably less apoptosis in the spleens and had a higher survival rate after burns. Heparin did not significantly decrease apoptosis in the spleen and the mortality rate in IL-1 R-/- mice after burns.

CONCLUSIONHeparin inhibits burn-induced apoptosis of the spleen cells by suppressing IL-1 expression in mice.

Animals ; Apoptosis ; drug effects ; Burns ; drug therapy ; metabolism ; Heparin ; therapeutic use ; Interleukin 1 Receptor Antagonist Protein ; pharmacology ; Interleukin-1 ; metabolism ; Male ; Mice ; Mice, Knockout ; Receptors, Interleukin-1 ; metabolism ; Spleen ; drug effects ; metabolism

OBJECTIVETo investigate Siwu decoction and its composite drugs on the blood-deficiency model mice induced by acetylphenyhydrazine and cyclophosphamide.

METHODAcetylphenyhydrazine and cyclophosphamide were used to copy the blood-deficiency model mice. Automatic hematology analyzer was used to test the peripheral hemogram. Weighting method was used to test the liver index and spleen index; Kits for ATPase test was used to test the activities of Na+ - K+ - ATPase/Ca2+ - Mg2+ - ATPase in erythrocyte membrane. Flow cytometry was used to test the bone marrow cells' cell cycle.

RESULTAngelicar Sinensis Radix and Paeoniae Radix Alba had the most effective activity on the peripheral hemogram. Paeoniae Radix Alba, the drug pair including Angelicar Sinensis Radix and the drug- group including Paeoniae Radix Alba had the most effective activity on the liver index. All the drugs, drug-pairs, drug-groups and the formula had effect on the spleen index. To the activity of Na+ - K+ - ATPase in erythrocyte membrane, Rehmanniae Radix Praeparata, the drug-pairs and drug-groups including Angelicar Sinensis Radix exhibited the most effective activity. All the drugs, drug-pairs, drug-groups and the formula had the protective effect on the damaged bone marrow cells.

CONCLUSIONSiwu decoction and its composite drugs all had effect on the blood-deficiency model mice, but the action intensity was different. Angelicar Sinensis Radix and Paeoniae Radix Alba exhibited the most effective activity to the protection of the blood-deficiency model mice.

Adenosine Triphosphatases ; drug effects ; metabolism ; Angelica sinensis ; Animals ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cyclophosphamide ; Disease Models, Animal ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Female ; Glucosephosphate Dehydrogenase Deficiency ; chemically induced ; drug therapy ; metabolism ; pathology ; Liver ; drug effects ; metabolism ; Mice ; Mice, Inbred ICR ; Paeonia ; Phenylhydrazines ; Protective Agents ; pharmacology ; Spleen ; drug effects ; metabolism

AIMTo investigate effects of hsBAFF synthesized in Escherichia coli on spleen B lymphocyte immune response and its intracellular free Ca2+ ([Ca2]i]) signaling in mice.

METHODSTwenty ICR mice, half males-half females, were chosen and randomly divided into a normal control group (n=10) and a hsBAFF treatment group (n-10). The mice in hsBAFF treatment group were given abdominal cavity injection of hsBAFF solution which was diluted with phosphate buffered saline (PBS) at dosage of 0.1 mg/kg body weight once each day for over eight days. The mice in control group were received abdominal injection of PBS at the same dose and frequency. Spleen B lymphocyte proliferation and its immune response to LPS stimulation in mice were evaluated using an MTT assay, and change of spleen B lymphocyte [Ca2+]i was assayed under a laser scanning confocal microscope.

RESULTSB lymphocyte proliferation and its immune response to LPS stimulation were significantly higher in hsBAFF-treated mice than in control mice (P < 0.05). The B lymphocyte [Ca2+]i fluorescence intensity in hsBAFF-treated mice maintained at a relatively high level fluctuation, and its average intensity was significantly higher to that of control mice (P < 0.01), but change rate of the intensity was lower compared to that of control group.

CONCLUSIONhsBAFF synthesized in Escherichia coli can enhance immune function in the body by increasing B lymphocyte proliferation and its immune response. hsBAFF-activated B lymphocyte function may be associated with increasing B lymphocytes [Ca2+]i.

Animals ; B-Cell Activating Factor ; immunology ; pharmacology ; B-Lymphocytes ; cytology ; drug effects ; immunology ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; Cell Proliferation ; drug effects ; Female ; Male ; Mice ; Mice, Inbred ICR ; Spleen ; cytology ; drug effects ; immunology

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism