四物饮调控巨噬细胞极化抑制食管癌的机制研究OA

Objective It is to observe the inhibiting effect of Siwu-Yin on esophageal cancer and its influence on macro-phage polarization,and to explore the mechanism of action.Methods ①A control group and a Siwu group were set up in the experiment.Eca-109 highly differentiated human esophageal cancer cells(hereinafter referred to as Eca-109 cells)were inoculated in 12-well plates.After 25 hours of culture,the Siwu group was treated with 150 μg/mL Siwu-Yin,while the control group received no additional intervention.After another 24 hours of culture,a cell scratch assay was performed,and the percentage of unclosed scratches was calculated at 24 and 48 hours after scratch.Using the same groupings and cul-ture methods,the expressions of tumor-associated macrophage polarization proteins[interleukin(IL)-17B,macrophage colony-stimulating factor(MCSF),macrophage colony-stimulating factor receptor(MCSF R),and IL-4 receptor alpha subunit(IL-4 Ra)]in the Eca-109 cells were measured and analyzed by a 640-protein microarray.②Thirty C57BL/6 mice were randomly divided into control group(n=8),a carcinogen-induced group(n=11)and Siwu group(n=11).The experimental period lasted for 32 weeks.During the experiment,the control group was given free drinking of drinking water;the carcinogen-induced group and Siwu group were given free drinking of 0.1 mg/mL solution of 4-nitroquinoline-1-oxide(4NQO)from the 1st week to 16th week of the experiment.Starting from the 17th week,the carcinogen-induced group was given free drinking of drinking water,while the Siwu group was given free drinking of Siwu solution at a dose of 4.55 g/(kg·d).At the end of the 32th week,the mice were anesthetized and sacrificed.The histopathological morpholo-gy of esophageal tissue of mice in each group was observed,and the macrophage polarization(F4/80+CD68 double-posi-tive cells were defined as M1-type macrophages,and F4/80+CD163 double-positive cells were defined as M2-type macro-phages)in esophageal and splenic tissues of mice in the carcinogen-induced group and Siwu group were measured by flow cytometry.Results At 24 h and 48 h after scratch,the scratch unclosure rate of Siwu group was significantly higher than that of the control group(P<0.05);the protein expressions ofIL-17B,MCSF,MCSF R,IL-4 Ra of the Siwu group were significantly lower than those of the control group at the same period,the differences were all significant(a difference time greater than1.2 or less than0.8 indicated a statistically significant difference).HE staining showed that the esophageal cancer tissues in the carcinogen-induced group exhibited infiltrative growth in the form of nests or strands,with fibrous tis-sue proliferation and inflammatory cell infiltration in the stroma;the nests volume of esophageal cancer was relatively smal-ler,the depth of infiltration was shallower,and the degree of surrounding inflammatory cell infiltration was lower in the Si-wu group than those in the carcinogen-induced group.Flow cytometry results showed that the macrophages in esophageal and spleen tissues of the Siwu group mainly were M1 type,those in the carcinogen-induced group were M2 type,further-more,the proportion of M1-type macrophages was significantly higher(P<0.05),while the proportion of M2-type macro-phages was significantly lower in the Siwu group than those in the carcinogen-induced group(all P<0.05).Conclusion Siwu-Yin can obviously inhibit the occurrence and development of esophageal cancer,which may be achieved by improving the polarization of tumor associated macrophages.