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Objective:To investigate the effects of berberine on Sprague Dawley (SD) rat retinal Müller cells cultured by high concentration glucose.Methods:The cultured SD rat retinal Müller cells were divided into normal-glucose group, high-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, and the cells were cultured in 5 mmol/L glucose, 25 mmol/L glucose, 25 mmol/L glucose+ 10 μmol/L berberine, and 25 mmol/L glucose+ 25 μmol/L berberine, respectively.After 72 hours cultured, cell apoptosis rate was detected by flow cytometry; the expressions levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the culture supernatant were detected by enzyme linked immunosorbent assay (ELISA); the L-glutamate-L-aspartate transporter (GLAST) and the related protein expression levels were detected by real-time fluorescence quantitative PCR; the expressions levels of GLAST, protein phosphatase magnesium-dependent 1A (PPM1A), nuclear factor-κB (NF-κB) and cleaved caspase-3 in the cytoplasm, and the expression level of NF-κB protein in the nucleus were detected by Western blot.Results:The cell apoptosis rate was (1.37±0.21)%, (17.67±1.17)%, (10.60±0.17)% and (5.57±0.35)% in the normal-glucose group, high-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, respectively, and the overall comparative difference was statistically significant ( F=375.97, P<0.01). The cell apoptosis rates in the high-glucose group was increased in comparison with those in the normal-glucose group ( P<0.01). The cell apoptosis rates in the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group were significantly reduced in comparison with that in the high-glucose group (both at P<0.01). And the cell apoptosis rate in the high-glucose+ 25 μmol/L berberine group was lower than that in the high-glucose+ 10 μmol/L berberine group ( P<0.01). ELISA results revealed that the overall comparative differences of the concentrations of TNF-α, IL-8 and COX-2 among the four groups were statistically significant ( F=28.36, 35.88, 41.59; all at P<0.01). The concentrations of TNF-α, IL-8 and COX-2 in the high-glucose group were significantly higher than those in the normal-glucose group ( P<0.01). Compared with the high-glucose group, the concentrations of TNF-α and COX-2 were significantly decreased in the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (both at P<0.05). The concentrations of TNF-α and IL-8 in the high-glucose+ 25 μmol/L berberine group were lower than those in the high-glucose+ 10 μmol/L berberine group (both at P<0.05). Real-time fluorescence quantitative PCR showed that the relative expression levels of GLAST mRNA in Müller cells among the four groups were statistically significant ( F=268.60, P<0.01). Compared with the normal-glucose group, the relative expression level of GLAST mRNA was significantly decreased in the high-glucose group ( P<0.01). In the high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group, the relative expression level of GLAST mRNA was significantly elevated compared with the high-glucose group (both at P<0.05). Western blot analysis results showed that the overall comparative differences of the GLAST, PPM1A, cleaved caspase-3, NF-κB in cytoplasm and NF-κB in nucleus among the four groups were statistically significant ( F=135.20, 156.98, 80.96, 128.07, 47.36; all at P<0.01). The relative expression levels of GLAST, PPM1A and NF-κB protein in cytoplasm in the high-glucose group were significantly lower than those in the normal-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (all at P<0.05). The relative expression levels of cleaved caspase-3 and NF-κB protein in nucleus in the high-glucose group were significantly higher than those in the normal-glucose group, high-glucose+ 10 μmol/L berberine group and high-glucose+ 25 μmol/L berberine group (all at P<0.05). Conclusions:High-concentration glucose can induce cell apoptosis and inflammatory response of SD rats retinal Müller cells in vitro.However, berberine can inhibit cell apoptosis and inflammatory response induced by high-concentration glucose via suppressing NF-κB translocation and transcription activity, and thereby inhibiting the expression of inflammatory cytokines.
RESUMO
Objective:To evaluate a new nano-carbon lymphatic tracing method to increase the number of lymph nodes acquired in patients with neoadjuvant chemotherapy for gastric cancer.Method:From Jan 2015 to Mar 2016, 159 patients with gastric cancer were recruited including 66 cases in study group receiving nano carbon injection under the mucosa layer one day before the operation, and 93 cases with intraoperative subserosal layer injection as control.Results:The average number of lymph nodes obtained in the study group was 47.0±14.7, while that in control was 38.0±14.5, P<0.05. The number of fibrotic lymph nodes obtained in the study group was 3.1 ± 1.9, compared with 3.0±1.8 in control, P>0.05. The number of black-stained lymph nodes in the former was 22.3±4.4, and the later was 14.7±4.8, P<0.05. The lymph nodes harvested in the first station in study group was 26.6±8.5, while that in the control group was 24.1±9.9, P>0.05. The lymph nodes obtained in the second station was 20.4±6.9 in study group, while in control was 13.8±5.7, P<0.05. Conclusions:The submucosal injection of nanocarbon one day before surgery increase the number of lymph nodes obtained in gastric cancer patients with neoadjuvant chemotherapy.