Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods ...Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods After cultured in vitro for 14 d, the rat organotypic midbrain slices were pretreated with different concentrations （0, 1, 3, 6 or 10 ng/mL） of LPS for 24 h followed by treatment with 100 ng/mL LPS for 72 h. The whole slice viability was detelmined by measurement of the activity of lactic acid dehydrogenase （LDH）. Tyrosine hydroxylase-immunoreactive （TH-IR） neurons and CD 1 1 b/c equivalent-immunoreactive （OX-42-IR） microglia in the slices were observed by immunohistochemical method, and tumor necrosis factor-α （TNF-α levels in the culture media were detected by enzymelinked immunosorbent assays （ELISA）. Results In the slices treated with 100 ng/mL LPS for 72 h, the number of TH-IR neurons reduced from 191± 12 in the control slices to 46±4, and the LDH activity elevated obviously （P 〈 0.01）, along with remarkably increased number of OX-42-IR cells and production of TNF-α （P 〈 0.01）. Preconditioning with 3 or 6 ng/mL LPS attenuated neuron loss （the number of TH-IR neurons increased to 126± 12 and 180± 13, respectively） and markedly reduced LDH levels （P 〈 0.05）, accompanied by significant decreases of OX-42-IR microglia activation and TNF-α production （P 〈 0.05）. Conclusion Low-dose LPS preconditioning could protect dopaminergic neurons against inflammatory damage in rat midbrain slice culture, and inhibition of microglial activation and reduction of the proinflammatory factor TNF-α production may contribute to this protective effect. Further understanding the underlying mechanism of LPS preconditioning may open a new window for treatment of Parkinson＇s disease.展开更多
OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS...OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS Ventral mesencephalic precursors from Ell inbred rat embryos and BMSCs from adult rats were cultured both separately and in co-culture. After a 7-day incubation in vitro, three conditioned culture media were obtained, termed VMP or common medium, BMSC medium, and BMSC±VMP medium. Ventral mesen- cephalic precursors cells were cultured in each of these media and the effects on proliferation and VMP differentiation were assessed. The relative yield of TH± cells was calculated and compared by immunocytochemical staining. RESULTS After a 7-day culture and induction of VMPs, the total cell counts were increased by （44.13±4.75）-fold （common）, （60.63±5.25）-fold （BMSC）, and （64.00±7.63）-fold （BMSC±VMP）. The proportions of TH＋ cells were （18.76±5.20）%, （23.49±4.10）%, and （28.08± 5.42）%, respectively, with statistically significant differences among the treatment groups. CONCLUSION BMSCs release factors that promote the proliferation of VMPs and facilitate the committed differentiation of VMPs into dopaminergic neurons.展开更多
基金the Foundation of Beijing Municipal Commission of Education,China (No.200410025011)
文摘Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods After cultured in vitro for 14 d, the rat organotypic midbrain slices were pretreated with different concentrations （0, 1, 3, 6 or 10 ng/mL） of LPS for 24 h followed by treatment with 100 ng/mL LPS for 72 h. The whole slice viability was detelmined by measurement of the activity of lactic acid dehydrogenase （LDH）. Tyrosine hydroxylase-immunoreactive （TH-IR） neurons and CD 1 1 b/c equivalent-immunoreactive （OX-42-IR） microglia in the slices were observed by immunohistochemical method, and tumor necrosis factor-α （TNF-α levels in the culture media were detected by enzymelinked immunosorbent assays （ELISA）. Results In the slices treated with 100 ng/mL LPS for 72 h, the number of TH-IR neurons reduced from 191± 12 in the control slices to 46±4, and the LDH activity elevated obviously （P 〈 0.01）, along with remarkably increased number of OX-42-IR cells and production of TNF-α （P 〈 0.01）. Preconditioning with 3 or 6 ng/mL LPS attenuated neuron loss （the number of TH-IR neurons increased to 126± 12 and 180± 13, respectively） and markedly reduced LDH levels （P 〈 0.05）, accompanied by significant decreases of OX-42-IR microglia activation and TNF-α production （P 〈 0.05）. Conclusion Low-dose LPS preconditioning could protect dopaminergic neurons against inflammatory damage in rat midbrain slice culture, and inhibition of microglial activation and reduction of the proinflammatory factor TNF-α production may contribute to this protective effect. Further understanding the underlying mechanism of LPS preconditioning may open a new window for treatment of Parkinson＇s disease.
基金This work was supported by grants from Natural Science Foundation of Jiangsu Province （No.BK2004043）
文摘OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS Ventral mesencephalic precursors from Ell inbred rat embryos and BMSCs from adult rats were cultured both separately and in co-culture. After a 7-day incubation in vitro, three conditioned culture media were obtained, termed VMP or common medium, BMSC medium, and BMSC±VMP medium. Ventral mesen- cephalic precursors cells were cultured in each of these media and the effects on proliferation and VMP differentiation were assessed. The relative yield of TH± cells was calculated and compared by immunocytochemical staining. RESULTS After a 7-day culture and induction of VMPs, the total cell counts were increased by （44.13±4.75）-fold （common）, （60.63±5.25）-fold （BMSC）, and （64.00±7.63）-fold （BMSC±VMP）. The proportions of TH＋ cells were （18.76±5.20）%, （23.49±4.10）%, and （28.08± 5.42）%, respectively, with statistically significant differences among the treatment groups. CONCLUSION BMSCs release factors that promote the proliferation of VMPs and facilitate the committed differentiation of VMPs into dopaminergic neurons.
