RESUMO
Objective To investigate influence of ginsenoside Rb1 on the proliferation and bioactivity of olfactory ensheathing cells (OECs).Methods OECs were primary cultured and purified from olfactory bulb of the adult SD rats.MTT assay was used to detect proliferation of OECs treated with ginsenoside Rb1 (intervention concentrations of 0,10,20,40,and 80 μg/ml and intervention time of 12,24,36,48,and 60 hours).Optimal concentration and intervention time of ginsenoside Rb1 was determined and performed in the succedent experiments.Purified cells were divided into blank control group and ginsenoside Rb1 group.RT-PCR was utilized to determine mRNA expressions of nerve growth factor (NGF),brain-derived neurotrophic factor (BDNF),glial derived neurotrophic factor(GDNF) and neural cell adhesion molecule (N-CAM) in the two groups.ELISA analysis was performed to measure secretion levels of NGF,BDNF and GDNF in the cultural supernatant.Results MTF analysis suggested ginsenoside Rb1 promoted proliferation of OECs with optimal effect at 20 μg/ml concentration for 48 hours (0.648±0.019,P ＜ 0.05).RT-PCR analysis demonstrated that mRNA expressions of NGF,BDNF,GDNF and N-CAM were significantly up-regulated in ginsenoside Rb1 group compared to those in blank control group (0.620 ± 0.011 vs 0.180 ± 0.011,0.511 ± 0.090 vs 0.293 ± 0.051,0.343 ± 0.042 vs 0.064 ± 0.005,0.839 ± 0.017 vs 0.717 ± 0.044) (P ＜ 0.05).ELISA analysis confirmed that secretions of NGF,BDNF and GDNF was increased in Rb1 group compared to those in blank control group (200.167 ± 8.361 vs 51.467 ± 3.815,156.700 ± 4.190 vs 96.500 ± 2.707,26.264 ± 5.864 vs 4.917 ± 10.894,P ＜ 0.05).Conclusion Ginsenoside Rb1 significantly promotes proliferation and bioactivity of OECs and hence benefits to spinal cord injury repair.
RESUMO
Florfenicol is a new type of broad-spectrum antibacterial that has been used in veterinary clinics. It shows immunosuppressive activity on the immune responses to ovalbumin (OVA) in mice. In the present study, florfenicol suppressed lipopolysaccharide (LPS)-stimulated splenocyte proliferation in a concentration-dependent manner in vitro and in vivo. BALB/c mice were immunized subcutaneously with OVA on days 1 and 4. Following the second immunization, mice were treated with a single daily oral dose of florfenicol (50, 100, and 200 mg/kg) for 10 consecutive days. On day 14, blood samples were collected to analyze OVA-specific IgG, IgG1, and IgG2b antibodies, and splenocytes were harvested to assess lymphocyte proliferation, CD3(+) T and CD19(+) B lymphocyte subsets. The results presented here demonstrate that florfenicol not only significantly suppressed Con A-, LPS- and OVA-induced splenocyte proliferation but also decreased the percentage of CD19(+) B cells in a dose-dependent manner and suppressed CD3(+) T cell at high doses. Moreover, OVA-specific IgG, IgG1 and IgG2b titers in OVA-immunized mice were reduced by florfenicol. These results suggest that florfenicol could suppress humoral and cellular immune responses in mice.
