OBJECTIVETo confirm the anticancer effect of total annonaceous acetogenins (TAAs) abstracted from Annona squamosa Linn. on human hepatocarcinoma.

METHODSThe inhibitory effect of TAAs was demonstrated in H22-bearing mice. The potency of TAAs was confirmed as its 50% inhibiting concentration (IC50) on Bel-7402 cell under Sulfur Rhodamine B staining. Both underlying mechanisms were explored as cellular apoptosis and cell cycle under flow cytometry. Mitochondrial and recipient apoptotic pathways were differentiated as mitochondrial membrane potential under flow cytometry and caspases activities under fluorescence analysis.

RESULTSThe inhibitory rate of TAAs in mice was 50.98% at 4 mg/kg dose. The IC50 of TAAs on Bel-7402 was 20.06 µg/mL (15.13-26.61µg/mL). Effective mechanisms of TAAs were confirmed as both of arresting cell cycle at G1 phase and inducing apoptosis dose- and time-dependently. Mitochondrial and recipient pathways involved in apoptotic actions of TAAs.

CONCLUSIONTAAs is effective for hepatocarcinoma, via inhibiting proliferation and inducing apoptosis.

Acetogenins ; chemistry ; pharmacology ; therapeutic use ; Animals ; Annona ; chemistry ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Humans ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Organ Specificity ; drug effects ; Spleen ; drug effects ; Thymus Gland ; drug effects ; Xenograft Model Antitumor Assays

Ginseng and its effective components are famous for their influence to enhance human immunity, regulate endocrine and antioxidant action. However, the different effects of different components are not clear. In this study, Wistar rats were used to study the effects of main components of ginseng, including total ginsenoside, panaxadiol saponins, panaxtrol saponin and ginseng polysaccharide. The results showed that the effects of panaxadiol saponins and ginseng polysaccharide on improving animal immune organ weight, plasma interleukin 2 (IL-2), interleukin 6 (IL-6), plasma gamma-interferon (IFN-γ), tumor necrosis factor alpha (TNF-α) were better than that of the other groups. Total ginsenoside and panaxtrol saponin can effectively increase the concentration of spleen NK cells (NKC) while panaxadiol saponins and ginseng polysaccharide can significantly increase the concentrations of rat plasma adrenocorticotrophic hormone (ACTH), corticosterone (CORT) and thyroid stimulating hormone (TSH). As for the effect of increasing organization nitric oxide (NO) and superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA), total ginsenoside is better than that of other groups. In brief, different components in ginseng possess different effects on enhancing immunity, regulating endocrine and resisting oxidation. Panaxadiol saponins and ginseng polysaccharide are better in enhancing immune, and total ginsenoside shows advantages in resisting oxidation and stress.
Adrenal Glands ; drug effects ; growth & development ; Adrenocorticotropic Hormone ; blood ; Animals ; Brain ; drug effects ; metabolism ; Corticosterone ; blood ; Ginsenosides ; pharmacology ; Glutathione ; metabolism ; Immune System ; drug effects ; physiology ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Killer Cells, Natural ; drug effects ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Organ Size ; drug effects ; Panax ; chemistry ; Polysaccharides ; pharmacology ; Random Allocation ; Rats, Wistar ; Saponins ; pharmacology ; Spleen ; drug effects ; growth & development ; Superoxide Dismutase ; metabolism ; Thymus Gland ; drug effects ; growth & development ; Thyrotropin ; blood

AIM: To improve the absorption of thymopeptides (TH) by preparing sodium deoxycholate/phospholipid-mixed nanomicelles (SDC/PL-MMs). METHODS: TH-SDC/PL-MMs were prepared by a film dispersion method, and then evaluated using photon correlation spectroscopy (PCS), zeta potential measurement, as well as their physical stability after storage for several days. Furthermore, in situ intestinal single-pass perfusion experiments and pharmacodynamics in immunodeficient mice were performed to make a comparison with TH powders and the control drug in absorption properties. RESULTS: A narrow size distribution of nanomicelles, with a mean particle size of (149 ± 8.32) nm and a zeta potential of (-31.05 ± 2.52) mV, was obtained. The in situ intestine perfusion experiments demonstrated a significant advantage in absorption characteristics for TH compared to the other formulations, and oral administration of TH-SDC/PL-MMs potentiated an equivalent effect with i.h. TH in pharmacodynamic studies in immunodeficient mice. CONCLUSIONS TH-SDC/PL-MMs prepared by a film dispersion method are able to improve the absorption of TH. SDC/PL-MMs might be a good approach for the more effective delivery of drugs like TH.
Animals ; Chemistry, Pharmaceutical ; Deoxycholic Acid ; chemistry ; Drug Carriers ; chemistry ; Drug Stability ; Mice ; Micelles ; Particle Size ; Peptides ; administration & dosage ; chemistry ; pharmacokinetics ; Phospholipids ; chemistry ; Rats ; Rats, Wistar ; Thymus Gland ; chemistry

Blood deficiency model of mice was copied by subcutaneous injection with 200, 100 and 100 mg x kg(-1) (0.01 mL x g(-1)) acetyl phenylhydrazine (APH) at the frist, fourth, and seventh days. Mice in each group were perfused with different extracted parts of Angelica sinensis (drug dosage was 2.4 g x kg(-1)) at the tenth day, once a day for 10 days. Then compare the influence of different extracted parts of Angelica sinensis to RBC, Hb, PLT and thymus, spleen and weight changes of blood deficiency mice. The peak areas of each common peak from HPLC fingerprint were associated with the date of replenishing blood pharmacodynamics efficacy by using gray relation statistic, which was used to research the chromatogram-pharmacodynamics relationship. The results showed that the part of DSC has the better effect in replenishing blood. The contribution degree of the DSC to replenishing blood of each component were determined by correlation size, and ferulic acid made the largest contribution, but contribution of other components should not be ignored. In this paper, we research the relationship of the HPLC fingerprint and spectrum activity relationship, determine the material basis of the DSC for replenishing blood, and provide effective way to represent the spectral correlation effect.
Angelica sinensis ; chemistry ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Erythrocytes ; cytology ; drug effects ; Female ; Hematologic Diseases ; drug therapy ; Humans ; Male ; Mice ; Spleen ; drug effects ; Thymus Gland ; drug effects

To study the toxicity-decreasing and synergistic effect of intracellular and extracellular polysaccharides from Phellinus igniarius on S180 mice. The PIP and PIE were extracted from the products of liquid submerged fermentation of P. igniarius. Transplanting S180 mice tumor models were established so as to observe the changes in tumor inhibiting rate, indexes of the spleen and thymus, body weight, peripheral blood cells and IFN-gamma levels when CTX was used alone and when used in combination with the PIP and PIE from P. igniarius. The results indicate that the PIP and PIE from P. igniarius can increase the activity of body immunity, attenuate the toxicity of CTX as well, and improve the anti-tumor effects.
Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Basidiomycota ; chemistry ; classification ; isolation & purification ; Body Weight ; drug effects ; Cyclophosphamide ; pharmacology ; Disease Models, Animal ; Drug Synergism ; Female ; Interferon-gamma ; blood ; Male ; Medicine, Chinese Traditional ; Mice ; Neoplasm Transplantation ; Plant Extracts ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Sarcoma 180 ; drug therapy ; pathology ; Spleen ; pathology ; Thymus Gland ; pathology

OBJECTIVEThrough establishing different blood deficiency animal model, to evaluate enriching blood effect changes of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma and each single herb, and to explore the effect characteristics of their compatibility.

METHODThree different methods of acetyl phenylhydrazine (APH) hemolytic method, cyclophosphamide (CTX) chemical damage method, APH-CTX complex method were used respectively to copy different blood deficiency model mice. Changes of orbit blood routine, thymus index, spleen index and ATPase activity of red cell membrane of model mice were tested.

RESULTCompared with normal group, all indexes had significant differences in three model mice. The drug pair and each single herb had significant impact on most indexes of the APH-CTX complex model mice, and on the individual indexes of APH hemolytic model mice and CTX chemical damage model mice. Therefore, APH and CTX complex blood deficiency model was more suitable for the enriching blood mechanism study of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma. Compared with the single herb of Angelicae Sinensis Radix and Chuanxiong Rhizoma, the drug pair of them had presented enriching blood effect at different extent with strengthening trend in regulating the invigorating blood indexes, immune organs and energy metabolic enzymes.

CONCLUSIONThe results of this research have provided scientific basis for revealing the mutual promotive composition law of the drug pair of Angelicae Sinensis Radix and Chuanxiong Rhizoma, and responded effectively the mult-link and mult-target effect characteristics of Chinese medicine bio-effect, to offer reference for the bio-effect research of the complicated substance group of Chinese medicine and traditional Chinese medicine formulae, and to supply demonstrative reference for researching the formulae compatibility law which takes the single drug-drug pair-formulae as main line.

Administration, Oral ; Angelica sinensis ; chemistry ; Animals ; Ca(2+) Mg(2+)-ATPase ; drug effects ; metabolism ; Cyclophosphamide ; pharmacology ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; chemistry ; Erythrocytes ; drug effects ; enzymology ; Female ; Hematologic Diseases ; drug therapy ; etiology ; Hemoglobins ; drug effects ; Leukocytes ; drug effects ; Medicine, Chinese Traditional ; Mice ; Models, Animal ; Phenylhydrazines ; pharmacology ; Plant Roots ; chemistry ; Random Allocation ; Rhizome ; chemistry ; Sodium-Potassium-Exchanging ATPase ; drug effects ; metabolism ; Spleen ; drug effects ; immunology ; Thymus Gland ; drug effects ; immunology

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

OBJECTIVETo study effects of polysaccharide of Radix Ranunculi Ternati (PRT) on immunological function and anti-oxidation activity of mouse.

METHODCell proliferations of splenocyte, thymocyte and peritoneal macrophage were measured by MTT colorimetry. The phagocytic function of peritoneal macrophage was measured by neutral red colorimetric method. The disoxidation power of PRT was measured by Prussian blue method. The clearing effect of PRT on hydroxyl radical was measured by salicylic acid capture method. The clearing effect of PRT on superoxide anion free radical was measured by pyrogallol auto oxidation method.

RESULTPRT among 25-400 mg x L(-1) could enhance thymocytes and spleen lymphocyte proliferation and macrophage phagocytosis. PRT(200 mg x L(-1)) has the strongest macrophage proliferation. PRT in different concentration has shown some disoxidation effects. PRT in 8 g x L(-1) has nearly the same ability of clearing x OH by Vit C with the same concentration. The clearance rate of PRT on O2*- is 95.39%.

CONCLUSIONPRT can enhance the cell proliferation capability of thymocytes, spleen lymphocytes and peritoneal macrophages. PRT can enhance macrophage phagocytosis in a dose-response relationship. PRT has saome disoxidation power and strong ability of clearing x OH and O2*-.

Animals ; Antioxidants ; analysis ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Macrophages, Peritoneal ; cytology ; drug effects ; immunology ; Male ; Mice ; Oxidation-Reduction ; drug effects ; Phagocytosis ; drug effects ; Polysaccharides ; immunology ; pharmacology ; Ranunculus ; chemistry ; Spleen ; cytology ; drug effects ; immunology ; Thymus Gland ; cytology ; drug effects ; immunology

The aim of this study was to investigate the effect of evodiamine on the proliferation and the immune function of thymocytes and splenocyte of mice from three germlines, which were 8 weeks old masculinity BALB/c, C57BL/6 and F1 hybridization mice. Cells of thymus and spleen were harvested and prepared as unicellular suspension. Cell proliferation was detected by MTT method, while the concentration of IL-2 was detected by ELISA, mRNA levels of bcl-2 and cdk2 in cells treated with evodiamine were detected by RT-PCR, the apoptosis rate and intracellular reactive oxygen species (ROS) concentration were analyzed by FCM, and the protein levels of BCL-2, CDK2 and BAX were determined by fluorescence microscope. The results indicated that at 0.5, 0.75 and 1 micromol/L, evodiamine inhibited the proliferation and externalization of thymocytes and splenocytes stimulated with ConA (p < 0.05). At 0.75 micromol/L, evodiamine inhibited the secretion of IL-2, decreased the mRNA level of bcl-2 and cdk2, and induced apoptosis of thymocytes and splenocytes (p < 0.05). Intracellular ROS concentration increased significantly after treatment with evodiamine for 12 hours (p < 0.05). The death rate increased at a prolong period of time. After treatment with evodiamine for 24 and 48 hours, the cells were divided into two groups, one of which was negatively stained by 2 7-dichlorofluorescein (DCF), which indicated that ROS level decreased significantly in the dying cells. It is concluded that evodiamine inhibits proliferation and induces apoptosis of thymocytes and splenocytes from different germline mice, and at the same time decreases secretion of IL-2 through down-regulating bcl-2 and cdk2 levels.
Adjuvants, Immunologic ; pharmacology ; Animals ; Cyclin-Dependent Kinase 2 ; genetics ; metabolism ; Drugs, Chinese Herbal ; chemistry ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Plant Extracts ; isolation & purification ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Quinazolines ; isolation & purification ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Species Specificity ; Spleen ; cytology ; immunology ; Thymus Gland ; cytology ; immunology

The study was purposed to analyze the relationship between the content of T-cell receptor excision DNA circles (TREC) and bcr-abl mRNA levels in CML patients and to evaluate the prognostic significance of recent thymic output function detection in patients with chronic myelogenous leukemia (CML). Quantitative detection of TREC and bcr-abl fusion gene transcripts in peripheral blood from 15 CML patients were preformed by real-time PCR. The change of bcr-abl levels in 6 patients was followed-up for two years. The results showed that there was no significant correlation between TREC and bcr-abl mRNA levels in peripheral blood from CML patients for the first attack. Patients who had higher TREC at diagnosis had a larger reduction of bcr-abl after 2 years of follow-up. While out of 2 patients who underwent haemopoietic stem cell transplantation (HSCT), one patient with higher level of TREC before transplantation was confirmed to express undetectable level of TREC by three consecutive detections after transplantation, other one patient was identified to express low level of bcr-abl. It is concluded that high thymic output function in CML patients can be beneficial for killing the residual CML cells.
Adolescent ; Adult ; Aged ; Female ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; Gene Rearrangement ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-abl ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcr ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Antigen, T-Cell ; analysis ; immunology ; T-Lymphocytes ; chemistry ; immunology ; Thymus Gland ; immunology