| Purpose:1.To investigate the effect of the supernatant of peripheral blood mononuclear cells(PBMCs)pretreated with Bozhi glycopeptide on the morphology of gastric cancer cells AGS.2.By the co-culture of PBMCs with gastric cancer cells AGS,we set the time factors and the grouping factors to investigate the effect of PBMCs pretreated with Bozhi glycopeptide on the relative proliferation activity of gastric cancer cells AGS.3.The wound healing assay was used to determine the effect of the PBMCs supernatant pretreated with Bozhi glycopeptide on the migration of gastric cancer cells AGS.4.To predict the possible targets and pathways of Bozhi glycopeptide on PBMCs with the network pharmacological methods,which provided a theoretical basis for the follow-up clinical research.Methods:1.The PBMCs supernatant pretreated with Bozhi glycopeptide was taken as a grouping factor and was cultured for 24 h.After 24 h,the IPP 6.0 software was used to measure the area,nuclear area,minimum feret,maximum feret,average feret,spindle cell area and other morphological parameters of gastric cancer cells AGS,and the statistical differences of the morphological parameters between different groups were compared.2.The PBMCs pretreated with Bozhi glycopeptide was taken as a grouping factor and a co-culture of PBMCs with AGS was performed.The proliferation activity data of AGS cells at 4h,8h,12 h,24h and 48 h were measured,and the anti-cancer activity data of the PBMCs with or without the pre-incubation of Bozhi glycopeptide were compared.3.The AGS cells were divided into two groups for intervention.The experimental group was a culture of PBMCs supernatant intervened with 150mg/L Bozhi glycopeptide.The control group was cultured with PBMCs supernatant untreated by Bozhi glycopeptide,but the wound healing was performed.By taking photos at the same 10 x field of vision at 0h,12 h and 24 h and using the IPP 6.0 software to process the photos,the migration distance of AGS cells was measured and the mobility was calculated.4.After reviewing literature,the active ingredients of Bozhi glycopeptide were screened and selected.Based on the network pharmacology analysis platform,the final prediction and screening on the main targets of Bozhi glycopeptide were completed through target function and pathway analysis.Results:1.The cell morphology experiment showed that the ratio of nucleus to cytoplasm,the spindle cell area,minimum Feret,maximum Feret and average Feret of AGS cells in the experimental group were all lower than those of the control group(t =-6.875,-8.102,-6.459,-8.191,-9.511)The P values of five cell morphological indicators were all less than 0.01,and the differences were statistically significant.2.The P value of the interaction between the grouping factor of the PBMCs pretreated with Bozhi glycopeptide and the time factor of the gastric cancer cell culture was less than 0.05,indicating that there was interaction between the grouping and the time factors,and the difference was statistically significant.In terms of the effect of the PBMCs pretreated with Bozhi glycopeptide on the relative proliferation activity of gastric cancer cells,the P value was less than 0.05,and the difference was statistically significant.The P value between the time factor and the grouping factor was less than 0.05,and the difference was statistically significant.The curve of the relative proliferation activity of AGS cells showed that the overall data of the relative proliferative activity of the experimental group,namely the AGS cells intervened with the PBMCs pretreated with Bozhi glycopeptide,was lower than that of the control group.3.The repeated measurement analysis of the average width of the scratch area showed the interaction between the grouping factor and the time factor,F=1.045,P=0.395,and indicated that there was no interaction between the two factors.The F value of the factors between groups of the PBMCs supernatant pretreated with Bozhi glycopeptide was 11.277,P<0.05,and the difference was statistically significant,indicating that the PBMCs supernatant pretreated with Bozhi glycopeptide significantly inhibited the migration of AGS cells.The F value of the time factor was 64.243,P<0.05,and the difference was statistically significant,indicating that the proliferative activity of AGS cells varied during different action time periods.The two-way ANOVA on the migration of gastric cancer cells AGS showed that the P value of interaction was 0.7323,indicating that there was no interaction between the treatment factor and the time factor of the PBMCs supernatant pretreated with Bozhi glycopeptide.The F value of the comparison between groups was 12.96 and the P value was 0.0070,indicating that the PBMCs supernatant liquid pretreated with Bozhi glycopeptide could significantly inhibit the mobility of AGS cells.The F value and P value of the time factor were 23.07 and 0.0014 respectively.The results showed that the mobility of AGS cells was different at different time points of culture.After 12 h incubation of the PBMCs supernatant liquid,the cell mobility data of the control and the experimental groups were 0.6220 and 0.3801 respectively,and the difference between them was-0.2419.The 95% confidence interval was(-.4800 ~-0.003872),t value 2.796,P<0.05.The difference was statistically significant.After 24 h incubation,the cell mobility data of the control and the experimental groups were 0.8942 and 0.6956,respectively.The difference was-0.1986,95% confidence interval(-0.4367 ~ 0.03947),t value 2.295.The P value was greater than 0.05.The mobility of the 24 h treatment group was lower than that of the control group,but the difference was not statistically significant(P>0.05).4.The function and pathway of ganoderpurine,one of the ingredients of Bozhi glycopeptide,were predicted.The cell composition,molecular function and biological process were also predicted,and it was found that the pathway that promoted cell proliferation was related to the network pathway of Glypican-1.Conclusions:1.Bozhi glycopeptide can regulate the function of PBMCs,further affecting the various morphological indicators that inhibit AGS cells and inhibiting the cell proliferation and migration.2.The network pharmacological analysis showed that Bozhi glycopeptide injection could regulate the PBMCs of gastric cancer patients,and then inhibit the cell proliferation and migration,and it may play a role through various pathways such as Glypican-1 network pathway... |
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