Kinetin (Kn) is a cytokinin growth factor that exerts several anti-aging and antioxidant effects on cells and organs. To investigate the mechanism underlying apoptotic events in aging cells induced by D-galactose (D-gal), we examined the effect of Kn delivered via nuchal subcutaneous injection on D-gal-induced aging and apoptosis in rats. Our results showed that interleukin (IL)-2 levels and mitochondrial membrane potential (DeltaPsim) were decreased by Kn in aging rats while IL-6 production and apoptosis increased. In addition, the expression of anti-apoptotic Bcl-2 was low while that of Bax was high in the aging group. After treated with Kn, compared with aging group, there showed obvious difference in Kn group with elevated IL-2, proliferation index, Bcl-2, DeltaPsim and decreased IL-6 and Bax in splenic lymphocyte. Based on these results, we concluded that Kn can effectively protect the rat spleen from aging, apoptosis, and atrophy.
Aging/drug effects/physiology ; Animals ; Apoptosis/drug effects/*physiology ; Female ; Galactose/*pharmacology ; Interleukin-6/physiology ; Interleukins/physiology ; Kinetin/pharmacology/*physiology ; Male ; Membrane Potential, Mitochondrial/drug effects/physiology ; Rats ; Spleen/*cytology/drug effects/physiology

The study purpose was to explore whether dichloromethylene diphosphonate (Cl(2)MDP)-loaded gelatin particles can induce the depletion of macrophage in reticuloendothelial system of liver and spleen or can depress the immunity of macrophage in SD rat models of immune thrombocytopenic purpura (ITP) to treat the ITP rats. New Zealand rabbits were immunized with platelets of SD rats to prepare rabbit anti-rat platelet serum, and the serum was intravenously injected into SD rats to produce the ITP model. In experimental ITP models, 150 microl of anti-platelet serum was intravenously injected into SD rats per 24 hours. The platelet counts maintained pathological level and were persistently less than 50 x 10(9)/L in the models during experiment process. The MTT test of macrophage RAW264.7 was carried out by means of Cl(2)MDP-loaded gelatin particles in vitro. After intravenous injection of a group dose of Cl(2)MDP-gelatin particles, the platelet counts of the rats were measured at the time of 4 hours, 24 hours, 48 hours, 72 hours and 96 hours, respectively, and bleeding times were detected in 24 hours. The results showed that Cl(2)MDP-loaded gelatin particles increased the platelet counts of ITP models to mean of 180 x 10(9)/L, a physiological level in 24 hours after injection, and kept this platelet level through whole process of 120 hours. Furthermore, rats pre-treated with Cl(2)MDP-loaded gelatin particles avoided the decrease of platelet counts significantly when they were injected anti-platelet serum. It is concluded that Cl(2)MDP-loaded gelatin particles restrain multiplication of macrophage RAW264.7, and promptly, effectively restore platelet counts of ITP models to physiological level in a dose dependent manner. So, the targeting therapy of drug-loaded gelatin particles offers a new idea and approach to treat ITP, and this strategy is worthy of further studies.
Animals ; Clodronic Acid ; administration & dosage ; Drug Carriers ; Drug Delivery Systems ; Gelatin ; administration & dosage ; Liver ; cytology ; drug effects ; Macrophages ; drug effects ; physiology ; Particle Size ; Purpura, Thrombocytopenic, Idiopathic ; therapy ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; drug effects

Thymopentin (TP5) and bursopentin (BP5) are both immunopotentiators. To explore whether the TP5-BP5 fusion peptide (TBP5) has adjuvant activity or not, we cloned the TBP5 gene and confirmed that the TBP5 gene in a recombinant prokaryotic expression plasmid was successfully expressed in Escherichia coli BL21. TBP5 significantly promoted the proliferation of thymic and splenic lymphocytes of mice. The potential adjuvant activity of the TBP5 was examined in mice by coinjecting TBP5 and H9N2 avian influenza virus (AIV) inactivated vaccine. HI antibody titers, HA antibodies and cytokines levels (IL-4 and IFN-γ) were determined. We found that TBP5 markedly elevated serum HI titers and HA antibody levels, induced the secretion of both IL-4 and IFN-γ cytokines. Furthermore, virus challenge experiments confirmed that TBP5 contributed to inhibition replication of the virus [H9N2 AIV (A/chicken/Jiangsu/NJ07/05)] from mouse lungs. Altogether, these findings suggest that TBP5 may be an effective adjuvant for avian vaccine and that this study provides a reference for further research on new vaccine adjuvants.
Adjuvants, Immunologic ; pharmacology ; Animals ; Antibodies, Viral ; blood ; Cell Proliferation ; drug effects ; Influenza A Virus, H9N2 Subtype ; drug effects ; physiology ; Influenza Vaccines ; immunology ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Lymphocytes ; drug effects ; Mice ; Oligopeptides ; immunology ; Orthomyxoviridae Infections ; drug therapy ; Recombinant Fusion Proteins ; immunology ; Spleen ; cytology ; Thymopentin ; immunology ; Thymus Gland ; cytology ; Vaccines, Inactivated ; immunology ; Virus Replication

OBJECTIVETo observe the evolution and differentiation of hepatic oval cells after transplanted into the spleens of homogenous rats, providing experimental data for treating hepatic failure with hepatic stem cells.

METHODSA two-step perfusion procedure was used to separate hepatic parenchymal cells from nonparenchymal cells. Then the suspension of nonparenchymal cells was centrifuged in Percoll gradients. The isolated cells were cultured, identified, and then transplanted into the spleens of homogenous rats undergone 2/3 hepatectomy.

RESULTSThe obtained cells were various in size with ovoid nuclei and inadequate cytoplasm. After 12 hours' culture, they revealed the characteristics of epithelial cells. Both the freshly isolated and cultured cells showed positive staining for cytokeratin 19 (CK19), OV6, alpha fetal protein (AFP), but negative for leucocyte common antigen (LCA). After intraspleenic transplantation into homogenous rats undergone partial hepatectomy, hepatic oval cells were differentiated into liver tissue-like structure including hepatocyte cords and bile ducts, and formed hepaticized spleen. But this kind of structure was not observed in the controls.

CONCLUSIONThe isolated rat hepatic oval cells show the biological characteristics of hepatic stem cells and can differentiate into hepatocytes and biliary epithelial cells under appropriate circumstances.

Animals ; Antigens, Differentiation ; analysis ; Cell Differentiation ; Cell Separation ; Cell Transplantation ; pathology ; Cells, Cultured ; Hepatectomy ; methods ; Hepatocytes ; drug effects ; physiology ; ultrastructure ; Liver ; cytology ; growth & development ; Liver Transplantation ; methods ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Spleen ; surgery ; Stem Cells ; drug effects ; physiology ; ultrastructure

A soluble factor which augments the expression of major histocompatibility complex class I (MHC I) antigens on a number of murine tumor cell lines, has been isolated from the culture supernatants of mixed lymphocyte reaction of spleen cells derived from C57B1/6, Balb/c and Swiss mice. The factor, termed MHC-augmenting factor (MHC-AF) has been partially purified by Sephadex G-100 column chromatography and reverse phase HPLC. MHC-AF activity is associated with an 18 kDa molecule. MHC-AF activity was resistant to pH 2.0 treatment and partially purified MHC-AF preparations did not have any activity in L929 cell/vesicular stomatitis virus (VSV) interferon bioassay system. Antibodies to IFN-gamma did not block the activity of MHC-AF. These results indicate that a MHC-AF distinct from IFN-gamma, is produced by mouse spleen cells undergoing a mixed lymphocyte reaction.
Animal ; Antibodies/pharmacology ; Chymotrypsin/metabolism ; Chymotrypsin/chemistry ; Comparative Study ; Concanavalin A/pharmacology ; Heat ; Histocompatibility Antigens Class I/metabolism* ; Histocompatibility Antigens Class I/drug effects ; Interferon Type II/pharmacology ; Interferon Type II/metabolism ; Interferon Type II/immunology ; Lymphocytes/physiology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Proteins/pharmacology* ; Proteins/metabolism ; Proteins/isolation & purification* ; Spleen/cytology ; Trypsin/metabolism ; Trypsin/chemistry ; Tumor Cells, Cultured/immunology ; Tumor Cells, Cultured/drug effects