Effect Of Gynostemma Pentaphyllum Polysaccharides On Mouse Peritoneal Macrophages

To investigate the immunoregulatory function and possible mechanism of GPP in the hosts,and to provide theoretical basis for the clinic application and further development,the phagocytic capability,proliferation,NO,LDH,TNF-a and IL-1β,CaM and GSa of peritoneral macrophages were detected by neutral red,Griess assay,CCK8,ELISA and RT-PCR.The results showed as below:1.Phagocytic capability of mice macrophages was promoted by the GPP.There was no significant difference of dose-effect relationship among various concentrations of GPP treatment.However,when 200 μg/mL of GPP was given,the phagocytic activity of macrophages reached maximum,and there was a significant difference compared to blank control(P<0.05).2.As for the cell proliferation of mice macrophages,compared with blank,different concentrations of GPP treatment could increase mice peritoneal macrophage proliferation.When 50 μg/mL of GPP was given,there was a significant increase of cell counts(P<0.05),but there was no significant difference among different GPP groups with the increase concentration of GPP.3.NO concentration in the culture supernatant of mice macrophages was significantly upregulated by different concentrations of GPP(P<0.05),and it was GPP dose-dependent.4.The levels of LDH in the culture supernatant of mice macrophages were significantly increase stimulated by both LPS and GPP,however,lacking a positive correlation.When 50 μg/mL and 100 μg/mL of GPP was given,there was a significant difference compared to the blank control,(P<0.05).5.As for the TNF-a production,compared to the blank control,it could significantly increase the TNF-a production when 50 μg/mL and above of GPP was given,(P<0.05).What’s more,it could increase followed by the increase of GPP concentration.The levels of TNF-a reached the maximum when the cells were stimulated with 100 μg/mL of GPP.However,the TNF-a production will tend to decrease as the GPP concentration continually increased.6.As for the IL-1β production,compared to the blank control,it could significantly increase the IL-1β production when 25 μg/mL and above of GPP was given,(P<0.05).What’s more,it could increase followed by the increase of GPP concentration.When 100 μg/mL of GPP was used,the level of IL-1βreached the maximum.The level of IL-1β was tend to decrease as the GPP concentration increased.7.The mRNA levels of CaM were altered by all concentrations of GPP in mice peritoneal macrophages.Compared to the blank control,it was significantly different when 25 μg/mL and above of GPP was given,(P<0.05).Amomg them the relative expression of CaM gene reached highest when the GPP dose was 100 μg/mL,The mRNA level of CaM was tend to decrease as the GPP concentration increased.8.The mRNA levels of GSa were significantly increased by all concentrations of GPP in mice peritoneal macrophages,(P<0.05).Especially treated with 25 μg/mL of GPP showing the strongest promotion to GSa expression.Conclusions:GPP has an activation effect on mice peritoneal macrophages.The cell proliferation of peritoneal macrophages can be promoted by GPP stimulation and therefore promotes the phagocytic capability of macrophages.GPP can also increase the secretion of NO and cytokines as LDH,TNF-a and IL-1β,and upregulated the gene expression of CaM and GSa in mice peritoneal macrophages.Based on the experimental results of this study,the effect was obvious when the GPP concentration was 50 μg/mL,and the best dose was 100 μg/mL.