Protective effect of Yiguanjian decoction against DNA damage on concanavalin A-induced liver injury mice model

OBJECTIVE:To investigate the inhibitory effect of Yiguanjian decoction(YD) on DNA damage in Concanavalin A(Con A)-induced liver injury mice model and to explain the possible mechanism.METHODS:Totally 120 male BALB/c mice were randomly divided into 6 groups,20 mice each:normal group,model group,Bifendate group,YD low dose group,YD middle dose group and YD high dose group.Except normal group,liver injury model induced by Con A was established.While modeling,each mouse in YD group was given YD(0.4 m L/20 g per day) by intragastric administration(0.13 g YD for YD low dose group;0.26 g for YD middle dose group;0.52 g for YD high dose group).Bifendate group was given Bifendate(0.2 gnd model·kg-1 grou·d-1) by gavage.Normal group ap were fed with same volume of physiological saline daily.After 8 weeks,the serum alanine transaminase(ALT)and aspartate transaminase(AST) were tested.The hematoxylin-eosin staining was used to evaluate the grade of liver inflammation and liver fibrosis stage.Hepatocellular DNA damage was detected by single cell gel electrophoresis technology.The protein expression of tumor necrosis factor-α(TNF-α),Bax and Mut T Homolog 1(MTH1) was detected by western blotting and enzyme linked immunosorbent assay.Bax m RNA and MTH1 m RNA were detected by Real-time Polymerase Chain Reaction(PCR).RESULTS:YD can improve the degree of liver inflammation and fibrosis in the liver of chronic hepatitis mice,the dose effect relationship is remarkable(P < 0.05).YD can reduce liver cell DNA damage.The difference between YD middle dose group and model group was statistically significant(P < 0.05).YD middle dose group had decreased the protein expression of TNF-α in the mice liver of immunological liver injury(P < 0.05).YD can increase the protein expression of Bax(P < 0.05).Compared with normal group,the protein expression of MTH1 was decreased(P < 0.05),but there was no statistical significance between YD group and model group(P >0.05).YD can increase the m RNA expression of Bax and MTH1(both P < 0.05).CONCLUSION:YD can effectively inhibit the DNA damage in immunological liver injury mice,the mechanism may be that it can decrease the TNF-αand increase the Bax and MTH1 expression.

Study on Molecular Mechanism of Liver Yang Ascending Syndrome in Hypertension

Objective: To investigate the molecular mechanism of Liver Yang Ascending Syndrome (LYAS) in hypertension. Methods: The plasma norepinephrine (NE), epinephrine (E) contents in patients with LYAS or Liver and Kidney Yin Deficiency Syndrome (LKYDS) of hypertension and normal controls were determined by high performance liquid chromatography-electrochemical detector (HPLC-ECD). The polymorphism of tyrosine hydroxylase (TH) gene in the three groups were analysed by Southern Blot assay. The polymorphism of TH and monoamine oxidation A(MAO-A), monoamine oxidation B(MAO-B) gene microsatellite in these groups were analysed by polymerase chain reaction (PCR)-SSCP. At the same time, the LYAS model was established by administering Aconitum preparation (AP) to spontaneous hypertension rats (SHR). The adrenal gland medulla of the experimental animal was examined for their TH expression at the protein level after ABC immuno-histochemical staining with the TH monoclonal antibody (McAb). The adrenal TH gene mRNA expression was demonstrated by in situ hybridization with the synthetized TH oligonucleic acid probe. The results of immuno-histochemistry and in situ hybridization were analyzed with the image analysis system (IAS). Results: The plasma NE and E contents in patients with LYAS were more obviously increased than those in LKYDS and normal controls. The TH gene amplified and the type A1 TH microsatellite D11S4046 was markedly higher in the LYAS group than those in the other groups. The TH mRNA protein expression in the adrenal tissue of the AP induced LYAS model in SHR elevated. Conclusion: The LYAS of hypertension has the characters as amplified TH gene and increased TH mRNA and protein expression, which suggested that the overexpression of TH is probably the mechanism of LYAS in hypertension.