Effect Of Gankang Granules On CD₄⁺, CD₈ +, IL-12 And Ifn-_γ In Model Of Immunological Liver Injury In Mice

Objective: To confirm the effect of Gankang granules on protecting liver from concanavalin A-induced immunological injury in mice, and its effect on the levels of CD₄⁺, CD₈⁺, IL-12 and IFN-γin concanavalin A (ConA)-induced immunological injury mice.Methods: sixty Kunming-strain mice were randomized into six groups including negative control group, model group, lentinus edodes mycelia polysacharide tablets(LEMP) treatment group, higher-, medium- and lower-dose of Gankang granules treatment groups. During a 7-d period, the mice in higher-, medium- and lower-dose of Gankang granules treatment groups were fed on Gankang granules at 12.5g/kg, 10.0g/kg and 7.5g/kg respectively, and the mice in LEMP treatment group were administered with LEMP at 0.06g/kg. Simultaneously, the mice in negative control group and model group were fed on equivalent saline water. After abrosia for 14 h, the mice, except negative control, were injected once with concanavalin A at 0.02g/kg for 6 hours, and then the levels of ALT, AST, IL-12 and IFN-γin the serum and the levels of CD₄⁺, CD₈⁺ in the blood were measured. Furthermore, the pathological morphology in liver tissues was observed using light microscope, and the index of liver, spleen and thymus were calculated.Results: 1) Hepatic function change: the level of ALT and AST in the serum in model group mice increased significantly. While, the level of ALT and AST in the serum in all Gankang granule groups mice decreased compared with the model group (P<0.05), especially that of decrease in the higher- and medium-dose groups. 2) CD₄⁺,CD₈⁺: Compared with the negative control, CD₄⁺ levels in the blood in the model group mice decreased (P<0.05), and CD₈⁺ levels in the blood in the model group mice increased (P<0.05). The levels of CD₄⁺ in the all Gankang granule treatment groups and LEMP treatment group were increased, while, the levels of CD₈⁺ were decreased. The effect between Gankang granule treatment and LEMP treatment on CD₄⁺ and CD₈⁺ was similar. 3) Pathology change: Model mice showed obvious degeneration of liver cells in water samples, point-like and necrosis of change. Compared with model group, the injured liver of mice in all treatment groups were ameliorated. Higher-dose of Gankang granule was the most effective one among three Gankang granule treatment groups. 4) Interleukin-12(IL-12) change: The level of IL-12 in the serum in model group mice increased significantly. Compared with model group, the level of IL-12 in all Gankang granule groups mice decreased significantly (P<0.05). 5) Interferon-γ(IFN-γ): The level of IFN-γin the serum in model group mice increased significantly. Compared with model group, the levels of IL-12 in all Gankang granule groups mice decreased significantly (P<0.05). 6) Visceral index: Compared with negative control, the index of liver, spleen and thymus of model group mice increased significantly (P<0.05), especially the index of liver, spleen(p<0.01). Compared with model group, the index of liver, spleen and thymus decreased in all treatment groups (P<0.05) in a dose-dependent manner.Conclusions: 1) Gankang granule has the capability of replenishing Qi to invigorate the spleen, invigorating the liver and tonifying the kidney, heat-clearing and detoxicating, resolving dampness and removing stasis, and can effectively prevent and treat immunological liver injury. 2) Liver is the target organ in ConA induced toxicity in vivo. The decrease in the level of CD₄⁺ T cell is corelated to ConA-induced liver injury. IFN-γand IL-12 play a major rool in ConA-induced liver injury. The liver injury induced by these two cytokines is one of the pathological mechanisms of immunological liver injury induced by ConA. 3) Gankang granule can ameliorate the immunological liver injury in a dose-dependent maner. 4) The effect of Gankang granule on protecting liver from injury may be related to the increasing in the lecvel of CD4 + , decrease in the level of CD₈⁺, IL-12 and IFN-γ.