Deficient Inducibility Of Hsp70 By Heat And Effects Of Qing Gan Li Shi Decoction On Patients With Gan Dan Shi Re Predominant Chronic Hepatitis B

Aim:1. To evaluate the effectiveness and safety of eliminating-the-moist-heat-in-liver (Qinggan Lishi) therapy of traditional Chinese medicine (TCM) for the treatment of moist-heat-in-liver-and-gall-bladder-predominant (Gandan-Shire-predominant) chronic hepatitis B, a randomized controlled trial of small-size sample was designed and conducted.2. The aim of the experiments in the 2nd part is to explore the heat-induced heat shock protein 70 expression and the alteration of cytokines level following heat treatment, thus to provide new information about the biological essence of moist-heat-in-liver-and-gall-bladder-predominant (Gandan-Shire-predominant) chronic hepatitis B(CHB).Methods:1. Patients who met the designed criteria were included and allocated randomly into 2 groups. Qinggan Lishi decoction, a traditional Chinese herb formula which was prescribed according to the principle of eliminating-the-moist-heat-in-liver therapy, was given to the experimental group combined with Lamivudine tablets. Compound Glycyrrhizin together with Lamivudine tablets were given to the controls. The administration duration of Qinggan Lishi decoction and Compound Glycyrrhizin tablets was 4 weeks. Visual analogue scores (VAS) of 7 independent symptoms and the general symptomatic VAS, ALT value, serum viral markers and HBV DNA quantitative value were recorded and compared between the 2 groups. Adverse events were monitored in the whole process of trial.2. Blood samples were obtained from 9 patients with Gandan-Shire-predominant CHB,9 patients with stagnancy-of-liver-Qi-and-deficiency-of-spleen-Qi-predominant (Ganyu-Pixu-predominant) CHB, and 9 normal controls, respectively, and were allocated into 3 groups by the syndrome type of traditional Chinese medicine (TCM). Peripheral blood mononuclear cells (PBMCs) were isolated from the blood samples and were given heat treatment. Some were left untreated in order to compare with those treated. Western-blot assay were performed to test the level of heat shock protein 70 in PBMCs and ELISA assay to test the expression of TNF-α, IFN-γand IL-4 in the supernatant of PBMCs.Results: 1. Seventy-five patients,34 for the experimental and 41 for the controlled, were included. One patient of experimental group was ruled out by the intention-to-treat analysis. The VAS on each single independent symptom and general symptomatic VAS and ALT value of both groups were all decreased after the initiation of treatment. At the 2nd,3rd, and 4th week, the experimental had significantly lower VAS on each of the 7 independent symptoms and general symptomatic VAS than the controlled had (P<0.05). Moreover, the ALT value of the experimental group was significantly lower than that of the controlled (P<0.05). At the 12th week of continuous administration of Lamivudine tablets, the differential value of HBV DNA quantity of experimental group was significantly higher than that of control (P<0.05). No severe adverse events occurred during the whole trial.2. The untreated PBMCs from 3 groups had similar expression capability of Hsp70 and 3 cytokines following 24h of normal incubation. However, the situation was different on the PBMCs treated by incubation at 42.5℃for 16h. Gandan Shire group showed significantly less expression of Hsp70 than the other 2 groups whereas had significantly higher level of TNF-αand IFN-y in the supernatant of PBMCs with heat treatment. Gandan Shire group also was tested significantly lower expression of IL-4 in the supernatant compared with that of the other 2 groups.Conclusions:1. The eliminating-the-moist-heat-in-liver (Qinggan Lishi) therapy of traditional Chinese medicine may have beneficial effect on improving the symptoms due to hepatitis and liver function for a particular group of patients with chronic hepatitis B who are supposed to be discriminated and identified in the TCM manner. However, the anti-viral effect remains to be verified by further clinical trials.2. The Gandan-Shire-predominant CHB shows evident deficiency of heat-inducibility of Hsp70 and imbalance of Thl/Th2 cytokine expression, which may convey specific characteristics of CHB with traditional Chinese medical Gandan Shire syndrome in terms of immunological and biological essence.