The Main Monomer Compounds In Vitro Anti-detoxifying Qi Fang Effect Of Endometrial Cancer

[Purpose]The macrophage can suppress tumor growth, however,tumor microenvironment can make macrophage polarize into tumor-associated macrophages (TAMs),which may promote tumor growth and metastasis. It is known that a certain number of TAMs exist in endometrial carcinoma (EC), and the number of TAMs is positively correlated with the malignant degree of endometrial carcinoma. Mr Zhiqiang Guo,who is a famous veteran teran doctors of TCM, summarized Toxin-Resolving and Qi-Boosting prescription according his several decades of clinical practice. By adding or subtracting some herbal medicines on the basis of the prescription, the clinical efficacy of the prescription in the treatment of endometrial carcinoma is effective. In this experiment, the inhibition effect of the main monomeric compounds, that are scutellaria barbata flavonoid, scutellaria barbata polysaccharides, spreading hedyotis herb polysaccharides and spreading hedyotis herb flavonoid, in the prescription on Ishikawa cell and TAMs, which incubated in conditioned medium of Ishikawa cell will be observed. The Toxin-Resolving and Qi-Boosting prescription’s anti-cancer effect in vitro experiment will be preliminary discussed. Accordingly, the experiment can provide certain theoretical and experimental basis for developing Traditional Chinese Medicine preparations against EC.[Method]1. The inhibition effect of4main monomeric compounds on Ishikawa cells after24h and48h was tested by MTT method.2. Lymphoma U973cells was incubated with conditioned medium of Ishikawa cells to establish the TAMs activation cell model in vitro.3. The expression of phenotypic molecules CD163and CD206in four groups was tested by flow cytometric assay.4. The level of cytokine IL-10in macrophage supernatant was tested by Elisa.[Result]1. Scutellaria Barbata flavonoid and Spreading Hedyotis Herb flavonoid (1.6mg/ml,800μg/ml,400μg/ml,200μg/ml,100μg/ml,50μg/ml,25μ g/ml,12.5μ g/ml) can suppress proliferation of Ishikawa cells. And compared with control group, the scutellaria barbata flavonoid (1.6mg/ml,800μ g/ml,400μ g/ml,p<0.001;200μ g/ml, p<0.05) groups have obvious inhibiting effect, difference was statistically significant. The spreading hedyotis herb flavonoid (1.6mg/ml,800μ g/ml,400μ g/ml, p<0.05) groups have obvious inhibiting effect, difference was statistically significant. The inhibition of cell proliferation is not obvious in Scutellaria barbata polysaccharides (5mg/ml,2.5mg/ml,1.25mg/ml,0.625mg/ml,0.3125mg/ml,156.25μg/ml,78.125μ g/ml,39.0625μ g/ml) and spreading hedyotis herb polysaccharides (10mg/ml,5mg/ml,2.5mg/ml,1.25mg/ml,0.625mg/ml,0.3125mg/ml,156.25μg/ml,78.125μ g/ml).2. The conditioned medium of Ishikawa cells can up-regulated the expression of phenotypic molecules CD163and CD206(p<0.05), difference was statistically significant. That is to say the conditioned medium of Ishikawa cells can make macrophage polarize into TAMs.3. Compared with control group, the scutellaria barbata flavonoid group and spreading hedyotis herb flavonoid group can down-regulated the expression of phenotypic molecules CD163and CD206(p<0.001), difference was statistically significant.4. Compared with control group, the scutellaria barbata flavonoid group (300μ g/ml) and spreading hedyotis herb flavonoid group (800μ g/ml) can obviously down-regulated the levels of cytokine IL-10(p<0.01), difference was statistically significant.[Conclusion]1.The scutellaria barbata flavonoid and spreading hedyotis herb flavonoid can inhibit proliferation of Ishikawa cells,2. Successfully established the tumor-associated macrophage model in vitro.3. The scutellaria barbata flavonoid and spreading hedyotis herb flavonoid play a role of antineoplastic effect in vitro by inhibiting TAMs’form.