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Selection of the Fraction with Anti-inflammatory and Immunsurpressed Activities from HGT and Its Immunopharmacological Study

Author: WuRongRong
Tutor: QiChunHui;ZhangYongXiang
School: PLA Military Academy of Medical Sciences
Course: Pharmacology
Keywords: HGT-5A rheumatoid arthritis acute inflammation cellular immune response material base
CLC: R96
Type: Master's thesis
Year: 2007
Downloads: 77
Quote: 1
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Abstract

Objective: To find out the fraction with anti-inflammatory and immunesurpressed activities form HGT, and evaluate its treatment on rheumatoid arthritis (RA). Methods: Croton oil-induced ear edema was used to assess the anti-acute inflammation effect. 2,4-Dinitrochlorobenzene(DNCB)-induced delayed hypersensitivity (DTH ) response was employed to observe the effect on cellular immune response. The adjuvant induced arthritis (AA) rat model and collagen II induced arthritis (CIA) mouse model were used to evaluate its treatment on RA. The content of prostaglandin E2 (PGE2) in paw tissue was mearsured by using ultraviolet spectrophotometer. The splenocyte prolifferation was dertermined with ~3H-TdR incorporation assay. The T lymphocyte subsets and percentage of regulator T cells (Treg) were analyzed by using FACS. ELISA was used to observe the serum levels of anti-collagen II antibody IgG and leukotriene B4 (LTB4). Luminex assay was used to determine the levels of TNF-α, IFN-γand IL-4 in the supernatant fluid for splenocyte culture. Results: (1) The fractions HGT-1, 2, 3 and 5 from HGT inhibited both croton oil-induced mouse ear edema and DNCB-induced DTH response. HGT-5, the water-soluble fraction, was better by general analysis. The main components of HGT-5 were polysaccharides. So HGT-5 A was obtained by deposition of HGT-5 with 80% ethanol, which was adopted to continue evaluating its treatment on RA. (2) HGT-5A (100～500)mg/kg (ig) inhibited croton oil-induced mouse ear edema and also surpressed DNCB-induced cellular immune response (50～100mg/kg). The splenocyte proliferation and IgG production of B lymphocytes were also decreased after incubation with HGT-5A (1～500μg/mL ) , in vitro. (3) HGT-5A (ig) 30 and 60mg/kg obviously decreased paw edema of AA rats. It also ameliorated the ankle joint tissue pathological changes of AA rats. HGT-5A increased the declined splenocyte proliferation and decreased the elevated levels of PGE2 in paw tissue and IgG in the supernatant fluid for splenocyte culture of AA rats. (4) HGT-5A (ig) 100 and 250mg/kg obviously decreased paw edema of CIA model mice. The ankle joint tissue pathological changes of CIA mice were also ameliorated after treatment of HGT-5A. HGT-5A increased the percentage of CD4~+CD25~+ Treg in spleen, resumed the ratio of CD4~+/CD8~+ T cells, reduced the contents of TNF-α, IFN-γand IgG produced by splenocytes of CIA mice. (5) The main components of HGT-5A were polysaccharides. Eight polysaccharides were purified from HGT-5A with DEAE-cellulose chromatography and gel chromatography, six of them were homogeneous polysaccharides and their monosaccharide composition was also analyzed by gas chromatography. The inhibition of the 6 homogeneous polysaccharides on mouse splenocyte proliferation, in vitro, was all equivalent to HGT-5A. Conclusions: HGT-5A has anti-inflammatory and immune regulatory activities , and it has obvious anti-rheumatoid arthritis effect. Polysaccharides may be the material base of this action.

Selection of the Fraction with Anti-inflammatory and Immunsurpressed Activities from HGT and Its Immunopharmacological Study

Abstract

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