Experimental Study on Auti-angiogenic Effect in Hepatocellular Carcinoma of Xiaoaiping Injection Alone and When Combined with Oxaliplatin and Rh-Endostain
|School||Nanjing University of Traditional Chinese Medicine|
|Course||Traditional Chinese Medicine|
|Keywords||hepatocarcinoma LCI-D20 Xiaoaiping injection oxaliplatin rh-endostatin angiogensis|
Objective:To observe the effects of Xiaoaiping injection alone and when combined with Oxal-iplatin and rh-Endostain(endostar) to inhibit the growth, metastasis and angiogensis of tumor in nude mice with the high metastatic human HCC transplanted in situ (LCI-D20),and investigate the mechanism of Xiaoaiping injection alone and when combined with chemotherapy and antia-ngiogenic therapy to cure hepatocarcinoma.Methods:Forty-eight nude mice with the high metastatic human HCC transplanted in situ(LCI-D20) were divided into six groups randomly,and with eight mice each group. Group A:Xiaoaip-ing injection alone (15ml/kg,d1-21),Group B:oxaliplatin alone(5mg/kg,q2d,d1-14),Group C:X-iaoaiping injection (15ml/kg,d1-21) combine with oxaliplatin (5mg/kg,q2d,d1-14), Group D: oxaliplatin(5mg/kg,q2d,d1-14) combine with rh-endostatin (endostar)(10mg/kg,dl-21),Group E: Xiaoaiping injection(15ml/kg,dl-21) combine with oxaliplatin(5mg/kg,q2d,dl-14) and rh-endo-statin (endostar)(10mg/kg,dl-21), Group F:normal saline(10ml/kg,dl-21) nagative control group.The six groups were injected intraperitoneally per day for 3 weeks from the 11th day after the tumor transplantation. Body weight was measured every 3 days.After three weeks treatment, the mice were killed to take blood, the tissue of transplantation tumor,liver and lung.The conce-ntration of serum VEGF,HIF-la were measured by ELISA,the expression of VEGF,HIF-la and MVD in transplantation tumor were detected by immunohistochemical staining and western blot.Results:1.The transplanted tumor weight in A, B, C, D and E groups are significantly lower than that of the nagative control group(P<0.05). The transplanted tumor weight in B, C, D, E groups are lower than that of the A group (P<0.01). According to Weeb coefficient, Xiaoaiping injection combine with oxaliplatin has addition effect, Xiaoaiping injection combine with oxaliplatin and rh-endostatin has synergism effect.Compared with two drugs groups, the three drugs group has synergism tendency, but the difference between them is not significant(P> 0.05).2. Xiaoaiping injection combined with oxaliplatin and rh-endostatin can decrease intrahepatic metastasis and lung metastasis.Although there are not significant difference between treatment groups and negative control group (P>0.05), intrahepatic metastasis of B,C, D and E groups are lower than negative control group, and lung metastasis have not hanppend in D and E groups.3. The concentration of serum VEGF in treatment groups are not different from negative control group (P>0.05). Xiaoaiping injection can decrease the expression of serum HIF-la,but the difference with negative control group is not significant(P>0.05). The expression of serum HIF-la of B,C,D,E groups are significant lower than negative control group (P<0.01),and the expression of serum HIF-la of B,C,E groups are lower than the A group (P<0.05) 4.In immunohistochemical staining, the expression of VEGF, HIF-la and MVD in transplantation tumor of treatment groups are lower than that of negative control group, but the difference between them is not significant(.P>0.05). Compared with negative control group,the expression of VEGF in D group and the expression of HIF-la in E group have significant difference(.P<0.01),and so is the expression of MVD in A,B,D,E groups(P<0.05).5. In western blot, Xiaoaiping injection alone or combined with oxaliplatin and rh-endostatin can down-regulate the expression of VEGF and HIF-la in transplantation tumor,the difference compared with negative control group is significant (P<0.01, P<0.05). but there are not significant difference among treatment groups(P>0.05).Conclusion:1. Xiaoaiping injection has weakly inhibitory effect on the liver tumor growth of LCI-D20 nude mice. Combination of Xiaoaiping injection and oxaliplatin can strengthen the effect to suppress tumor growth. While combined the third drug-rh-endostatin,the effect is more significant.2. Xiaoaiping injection down-regulates the expression of VEGF and HIF-la of LCI-D20 nude mice,suppress tumor growth through anti-angiogenic approach. The expression of VEGF and HIF-la have not change significantly when combined with oxaliplatin or both and rh-endostatin, this suggests more complex mechanism inside, and need to investigate further.3. This research provided the theory and experimental foundation for the combination of TCM with cytotoxic drugs and anti-angiogenic drugs to cure hepatocarcinoma.