Hospital of Putian University, Putian 351100, Fujian Province, China.
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China.
Neural Regen Res. 2013 Oct 25;8(30):2820-6. doi: 10.3969/j.issn.1673-5374.2013.30.004.
Wistar rats were intragastrically perfused with Chinese medicines used for tonifying the kidney. These included 0.180 g/mL of Herba Epimedii (Epimedium), Semen Cuscutae (Dodder Seed), or Herba Cistanches (Desertliving Cistanche), 0.04 mg/mL monoamine oxidase-B inhibitor selegiline, or distilled water for 14 consecutive days to prepare drug-containing serum or blank serum. MES23.5 cells in the logarithmic phase were cultured in media supplemented with 15% drug-containing serum for 24 hours, followed by incubation in culture solution containing 100 μmol/L H2O2 for 3 hours. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow tometry results showed that all drug-containing serums improved the survival rate of H2O2-injured MES23.5 cells, inhibited pro-apoptotic FasL and caspase-3 expression, promoted anti-apoptotic Bcl-2 expression. However, drug-containing serums had little influence on Fas expression in H2O2-injured MES23.5 cells. Enzyme-linked immunosorbent assay results showed that serum containing Herba Cistanches or Herba Epimedii increased the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor in injured MES23.5 cells; serum containing Semen Cuscutae only increased brain-derived neurotrophic factor expression; while expression of the above neurotrophic factors remained the same in cells treated with serum containing selegiline. These findings indicate that Chinese medicines used to tonify the kidney can protect nerve cells by regulating the expression of apoptosis-related factors and neuro-trophic factors in MES23.5 cells.
Wistar 大鼠连续 14 天经口给予补肾中药(淫羊藿 0.180g/mL、菟丝子 0.04mg/mL、肉苁蓉 0.180g/mL)、单胺氧化酶-B 抑制剂司来吉兰 0.04mg/mL 或蒸馏水制备含药血清或空白血清。对数生长期 MES23.5 细胞用含 15%含药血清的培养基培养 24 小时,然后在含 100μmol/LH2O2 的培养液中孵育 3 小时。MTT 法和流式细胞术结果表明,各含药血清均可提高 H2O2 损伤 MES23.5 细胞的存活率,抑制促凋亡 FasL 和 caspase-3 表达,促进抗凋亡 Bcl-2 表达。但含药血清对 H2O2 损伤 MES23.5 细胞 Fas 表达影响较小。酶联免疫吸附试验结果显示,含肉苁蓉或淫羊藿的含药血清可增加损伤 MES23.5 细胞神经生长因子、脑源性神经营养因子和胶质细胞源性神经营养因子的表达;含菟丝子的含药血清仅增加脑源性神经营养因子的表达;含司来吉兰的含药血清对上述神经营养因子的表达无影响。这些结果表明,补肾中药可通过调节 MES23.5 细胞凋亡相关因子和神经营养因子的表达来保护神经细胞。