| Objictive: To investigate the inner relationship between the disturbance of the nerve-endocrine -immunoregulation network and hepatic tissue injury, and parkly explore the immunological mechanisms of chronic viral hepatitis B; To observe the effects of Hai Zhu Yi Gan Mixture(HZYGM) on the nerve-endocrine -immunoregulation network of the mice with immunological hepatic injury ,and reveal the functional mechanisms of HZYG Mixture, and observe the different effects of the two therapies QU XIE method and FU ZHENG method on the nerve-endocrine -immunoregulation network of the mice with immunological hepatic injury, and provide the scientific basis for the clinic praction.Methods: 1. 64 SPF male mice were randomly divided into 8 groups:Normal control group,Model group day-11,day-18,day-25, HZYG group,QU XIE group,FU ZHENG group,Dexamethasone group ,8 mice each group.Except normal control group ,all mice were injected with Bacille Calmette-Guerin(BCG) 200 mg /kg from cauda vein at the first day of experiment,after 10 days, and were injected with Lipopolysaccharide(LPS) 0.4 mg /kg from cauda vein,then the mice model of hepatic injury were set up.From the first day , the normal control group and the model group were given normal saline 30 ml/kg,one time every day,for 25 days. HZYG group,QU XIE group,FU ZHENG group,Dexamethasone group were respectively given HZYG Chinese herbal, QU XIE Chinese herbal, FU ZHENG Chinese herbal,and dexamethasone 30 ml/kg,one time every day,for 25 days. All samples such as blood,liver,spleen,pituitarium were collected to be detected, 8 mice were respectively from the model group at the 11th/18th/25th day,the rest were at the 25th day.2. The changes of liver function and hepatic pathology were observed respectively in each group.3. The changes in substance P(SP), vasoactive intestinal peptide (VIP), PRL and ACTH contents in the pituitary gland and peripheral blood were determined by radio-immunoassary.4. PBMCs from the mice were separated routinely and stimulated by horboe-12-myristate-13-asetate(PMA)/Ionomycin/Monensin for 6 hours, the ratios of Thl/Th2 cells were detected by flow cytometry with two color intracellular cytokine staining .5. PBMCs from the mice were separated routinely and stimulated by ConA for 72 hours, the levels of IL-2, IFN-γ, IL-4, IL-10 in the supernatant were detected by ELISA.6. The mRNA transcriptive levels of IFN-γ and IL-4 in spleen cells of micewere detected by RT-PCR.7. Statistical methods: The data were expressed by x±s and were tested by t-test.Results: 1.Liver function ALT, AST in the model group were gradually higher than those of the normal group, with the increasing of hepatic injury; there was no pathological change in the normal control group, but there were liver cell swelling, ballooning degeneration, liver cell necrosis, inflammation ,inflammatory granuloma in the model group, obviously different from the normal control group (p<0.01).2. The changes in substance P(SP) and PRL contents in the pituitary gland and peripheral blood gradually increased, and were significantly higher than those of the normal at the 18th/25th day (P<0.05 or P<0.01) .3. The changes in VIP and ACTH contents in the pituitary gland and peripheral blood gradually decreased, and were significantly lower than those of the normal group at the 18th/25th day (P<0.05 or P<0.01) .4. Thl type cytokine IL-2, IFN-γ levels were gradually and significantly higher than those of the normal group at the 18th/25th day (P<0.05 or P<0.01) ; Th2 type cytokine IL-4, IL-10 levels were gradually and significantly lower than those of the normal group at the 18th/25th day a (P<0.05 or P<0.01) ;The mRNA expressions of IFN-y were gradually and significantly higher than those of the normal group (P<0.05 or P<0.01) , the mRNA expressions of IL-4 were gradually and significantly lower than those of the normal group at the 18th/25th day (P<0.05) ;The percentages of Thl cells were gradually and significantly higher than those of the normal group, the percentages of Th2 cells were gr... |
PDF Full Text Request