Anti-tumor Effect Of Chinese Medicine Compound Ganqingning Formula On H22 Tumor-bearing Mice And Its Mechanism

Objective To investigate the anti-tumor effects and mechanisms of chinese medicine compound Ganqingning(GQN)formula on H22 tumor-bearing mice.Methods Hepatoma H22 tumor-bearing models were randomized into normal group,model group cyclophsphamide(CTX)group(0.02 g/kg),low-,middle-,and high-dose GQN groups(55.36,27.68,13.84 crude drug g/kg),each group had 10 mice.Mice in the normal and model groups were administrated by gavage normal saline.corresponding drug were given in other groups for 10 days.In 24 hours after the last administration,all mice were sacrificed and the blood,tumor,thymus and spleen were taken.The tumor,thymus and spleen were weighed to calculate the tumor inhibition rate and the organ index.Blood routine exam,liver and kidney function index of mice were determined by automatic biochemical analyzer,including alanine amino-transferase(ALT),aspartate aminotransferase(AST),blood urea nitrogen(BUN),uric acid(UA)and creatinine(CR).HE staining method was used to stain the tumor tissues and the cell structure and pathological changes of tumor tissues were observed.Western blot was used to detect the protein expression levels of Bax,Bcl-2,Cyt-c and Cleaved-caspase3 in the tumors.Immunohistochemistry was used to detect the expressions of VEGF and MVD.qRT-PCR was used to detect the mRNA expression levels of Bax,Bcl-2,Caspase3 and VEGF in tumor tissue.Result(1)Compared with the model group,low-,middle-,and high-dose GQN groups could effectively inhibit the tumor growth with inhibitory rate22.17%,38.26%,47.39%,respectively(P<0.01).(2)Compared with the model group,The ALT,AST,BUN,UA,and CR levels remained unchanged(P>0.05).(3)Tumor pathology slices showed,compared with the model group,three doses of GQN groups could kill and reduce the tumor cells.(4)Compared with the model group,western blotting analyses showed three doses of GQN groups could increased the protein expressions of Cyt-c and decreased the protein expressions of Bcl-2(P<0.01).The middle-and high-dose GQN groups could increase the protein expressions of Bax and Cleaved-caspase-3(P<0.01).Compared with the model group,immunohistochemistry revealed obviously down-regulated expressions of VEGF in three doses of GQN groups(P<0.01,P<0.05).(5)Compared with the model group,qRT-PCR results showed that the mRNA expressions of VEGF was decreased in three doses of GQN groups(P<0.01).The mRNA expressions of Bcl-2 was decreased(P<0.01).the mRNA expressions of Bax and Caspase-3 were increasing in High-dose GQN group(P<0.01).Conclusion GQN formula has anti-tumor activity in H22 tumor-bearing mice.Its anti-tumor effect may be attributed to inducing the mitochondrial apoptosis and inhibiting angiogenesis.