Inhibitory Effects And Mechanism Research Of Hydroxysafflor Yellow A On H22 Cell Growth Of Hepatic Carcinoma In Mouse Model

Objective:Hydroxysafflor yellow A (HSYA), a flavonoid derived and isolated from traditional Chinese medicine Carthamus tinctorius L., possesses anti-tumor activity. Howerver, its effects on hepatocellular carcinoma (HCC) have not been investigated. Our study will reveal the mechanism of HSYA on inhibiting the growth of the hepatocellular carcinoma (HCC) in H22 tumor-bearing mice.Methods:1. After establishing the hepatoma model for 24 h, the mice were randomly divided into 5 groups. The vehicle-treated group received 0.9% normal saline via intraperitoneal injection. The positive control group was treated with sorafenib at a dosage of 50 mg/kg by intragastric administration. And the low, medium and high dose HSYA group received intraperitoneal injection at different dosages (1.125,2.25, 4.5 mg/kg), respectively. All these groups were treated one time daily for 14 days successively in a volume of 0.2 ml. After the last administration for 24 h, all the mice were sacrificed, and the weight change of mice, tumor growth inhibition rate and immune organ index were calculated.2. The mechanisms of HSYA on proliferation and apoptosis on tumor cell were investigated by immunohistochemical staining and flow cytometry.3. Immunohistochemical staining, enzyme-linked immunosorbent assays and western blotting were applied to measure the expression of angiogenesis related factors (VEGF, bFGF) and expression of ERK/MAPK (p-c-Raf, c-Raf, p-ERK1/2, ERK1/2).4. Western blot was used to evaluate the expression of NF-?B (p65, I?B and p-I?B) signaling pathway in H22 tumor-bearing mice among different groups.Results:1. Compared with saline group, HSYA could improve the tumor inhibitory rates, inhibit proliferation and promote apoptosis of tumor cells, especially the medium dose HSYA group, followed by the high dose HSYA group,but not better than the sorafenib-treated group.2. Compared with saline group and the sorafenib-treated group, HSYA could enhance the immunity of mice and have a dose-related beneficial influence on immune organ.3. HSYA could considerably inhibit the secretion of angiogenesis factors (VEGF A, bFGF) and restrain c-Raf and ERK1/2 phosphorylation.4. HSYA could block the activation of NF-?B and its nuclear translocation by down-regulating the expression of p65 in the nucleus, up-regulating p65 level in the cytoplasm, inhibiting I?B phosphorylation and cytoplasmic degradation of I?B-?.Conclusions:1. HSYA could improve the tumor inhibitory rates, inhibit proliferation and promote apoptosis of tumor cells in a certain concentration range.2. HSYA could inhibit the growth of tumor by enhancing the body immunity, inhibiting the angiogenesis of tumor and blocking ERK/MAPK and NF-?B signaling pathway in H22 tumor-bearing mice.