Study On The Inhibitory Effect Of Polysaccharide Extract From Gigantifolia Stapf On Ovarian Cancer Cells Based On The Antioxidant Effect

As a traditional Chinese medicine with Guangxi characteristics,the Ardisia Gigantifolia Stapf,polysaccharide is its main active ingredient.The objective of this study was to explore the optimal technological conditions,antioxidant and anti-ovarian cancer activity of polysaccharides from Ardisia Gigantifolia Stapf.Methods:1.Optimization of extraction technology and antioxidant activity of polysaccharides from Ardisia Gigantifolia Stapf:Ardisia gigantifolia Stapf,a genuine medicinal material from Guangxi Province,was selected as the test material.Single factor test and response surface methodology were used to optimize the extraction process of polysaccharide from Lonicerae japonica.Single factor test and response surface methodology were used to optimize the extraction process of polysaccharide from Ardisia gigantifolia Stapf.And the antioxidant activity of the extract was investigated.Five factors carried out single factor analysis and response surface test:extraction temperature,extraction time,extraction times,liquid-material ratio,alcohol precipitation ratio,and the best parameters of Ardisia gigantifolia Stapf polysaccharide extraction technology were obtained.The antioxidant activity was evaluated by DPPH radical,ABTS radical and hydroxyl radical scavenging test.2.The study on anti-ovarian cancer activity:The proliferation and apoptosis of SKOV3 AND A2780 cells were detected by MTT assay,wound healing assay,Hoechst 33258 staining,and flow cytometry,and the m RNA expressions of Bcl-2,Bax were detected by real-time PCR.Western blot was used to see the protein expressions of Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9,ERK,p-ERK,JNK,p-JNK,AKT,p-Akt and PI3K.Results:1.The results showed that the extraction rate of Ardisia gigantifolia Stapf polysaccharide was the highest under the conditions of extraction temperature80℃,extraction time 150 min,extraction times 4 times,liquid-material ratio1:21 g·m L^-1,and alcohol precipitation concentration 83%.Under the optimum conditions,the average extraction rate of Ardisia gigantifolia Stapf polysaccharide was 3.03%.In vitro,antioxidant test showed that the Ardisia gigantifolia Stapf polysaccharide had a specific scavenging effect on DPPH radical,ABTS radical and hydroxyl radical in the concentration range of0.0625-1.0 m Lmg·m L^-1.2.The acetate extract of Ardisia gigantifolia Stapf can can inhibit the proliferation of SKOV3 and A2780 cells in a concentration-dependent manner(IC₅₀was 64.23μg·m L^-1and 74.53μg·m L^-1).The extract up-regulated the expression of Bax m RNA,down-regulated the expression of Bcl-2 m RNA,up-regulated the expression of Bax,down-regulated the expression of Bcl-2 protein.Activation of Caspase-3,Caspase-8,and Caspase-9.decreased p-ERK/ERK and p-JNK/JNK ratio,decreased p-AKT/AKT ratio,and decreased expression of PI3K protein.Conclusions:It is feasible to optimize the process of Ardisia Gigantifolia Stapf Polysaccharide by single factor test combined with response surface methodology,and the extracted Ardisia gigantifolia Stapf polysaccharide has a particular antioxidant effect.The extract of Ardisia gigantifolia Stapf polysaccharide can inhibit the proliferation of SKOV3 and A2780 cells and induce apoptosis.The mechanism may be related to Bcl-2,Caspase family,MAPK,AKT/PI3K signaling pathway.