ABSTRACT
Objective:To observe the effects of Chinese herbal compound'Jisuikang'on the phagocytosis of microglia and the regeneration of injured neurons in co-culture system.Methods: Prepared drug serum of 'Jisuikang′ and isolated and identified the primary neuron and microglia.The neuron cells were induced apoptosis by glutamic acid and the microglia cells were predisposed by drug serum of'Jisuikang'.Then,the co-culture system of injured neurons and microglia cells was established.24 h and 96 h after co-culture,engulfment of neuron debris by microglia cells and regeneration of injured neurons were observed by immunofluorescence double labeling method.Results: 24 h after co-culture,middle and high dose of'Jisuikang' showed greater phagocytic percentage and phagocytic index than that of control.In comparison of LPS,high dose of'Jisuikang' showed no significant difference.96 h after co-culture,first grade of neuritis of middle and high dose of'Jisuikang' were more than that of control,and there were no significant difference in comparison of LPS.Neuritis' mean length per cell of middle and high dose of'Jisuikang' were larger than that of con-trol.Neuritis' mean length per cell of high dose of'Jisuikang' showed significant difference in comparison of LPS.Conclusion:Traditional Chinese medicine compound'Jisuikang'may enhance engulfment of neuron debris by microglia to improve local microenvi-ronment,which promote the repair and regeneration of injured neurons.
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Objective To investigate the effect of Chinese herbal compound"Jisuikang"on the phagocyto-sis of neuronal debris by microglial cells. Methods To prepare serum containing drugs of JSK and divide them into the low,middle and high dose groups,the blank serum group and LPS+blank serum group. BV2 was labeled by lentiviral vectors containing the green fluorescent protein gene (GFP). To establish the damage neuron model and mix injured neurons with the transfected microglia. To observe the situation of microglia which was affected by serum containing drugs devour the neuronal debris. Results The middle and high dose of JSK showed greater phagocytic percentage and phagocytic index than those of the control group(P<0.001). In comparison of LPS+blank serum group,no significant difference was found in the middle and high dose of JSK. However,to the phagocytic index, which was better than that of LPS+blank serum group(P<0.05). Conclusion JSK may enhance the engulfment of neuron debris by BV2,which could provide a better living environment for the growth of neurons.
ABSTRACT
Objective To investigate the effect of Chinese herbal compound"Jisuikang"on the phagocyto-sis of neuronal debris by microglial cells. Methods To prepare serum containing drugs of JSK and divide them into the low,middle and high dose groups,the blank serum group and LPS+blank serum group. BV2 was labeled by lentiviral vectors containing the green fluorescent protein gene (GFP). To establish the damage neuron model and mix injured neurons with the transfected microglia. To observe the situation of microglia which was affected by serum containing drugs devour the neuronal debris. Results The middle and high dose of JSK showed greater phagocytic percentage and phagocytic index than those of the control group(P<0.001). In comparison of LPS+blank serum group,no significant difference was found in the middle and high dose of JSK. However,to the phagocytic index, which was better than that of LPS+blank serum group(P<0.05). Conclusion JSK may enhance the engulfment of neuron debris by BV2,which could provide a better living environment for the growth of neurons.
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ABSTRACT Cyclin-dependent kinase-like 5 (CDKL5) is a protein kinase that is homologous to mitogen-activated protein kinases (MAPKs) and cyclin-dependent kinases (CDKs). Mutations in the CDKL5 gene cause X-linked infantile spasms and phenotypes that overlap with that of Rett syndrome, which is a neurodevelopmental disorder caused primarily by mutations in the methyl CpG binding protein 2 gene (MECP2). Previous studies in transfected cell lines showed that CDKL5 interacts with MeCP2 and DNA (cytosine-5)-methyltransferase 1 (Dnmt1). However, little is known about the relationships of CDKL5 with interacting proteins in primary neuronal cultures. In this study, we investigated the expression patterns of CDKL5, MeCP2 and Dnmt1, and their interaction in cultured rat cortical neurons. Using real-time PCR analysis, we found that CDKL5, MeCP2 and Dnmt1 have similar expression patterns at the mRNA level. In contrast, the expression patterns of those proteins at the protein level are different and could be inversely correlated, as shown by western blotting. Using co-immunoprecipitation, we further demonstrated that CDKL5 interacts with MeCP2 and Dnmt1 in primary rat cortical neurons. These data suggest that a functional link exists among CDKL5, MeCP2 and Dnmt1 during neuronal development and may provide further insight into the pathogenesis of Rett syndrome.
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Objective To observe the expression of peroxisome proliferator-activated receptor γ (PPARγ) in hippocampus neurons in rats after different time of neuron oxygen deprivation/oxygen supply, and to investigate the role of PPARγ in neuron ischemia reperfusion injury.Methods One day old newborn SD rats were chosen. Primary cultured hippocampus neurons were used to establish neuron ischemic reperfusion model in vitro by oxygen and glucose depriving 15 minutes and supplying again, and then the neuron structure was observed by transmission electron microscope of JEM-200EX.The expression of PPARγ mRNA and protein were detected by RT-PCR and Western blot, respectively.Results Neuron structure was damaged after neuron oxygen deprivation/oxygen supply. There was no significant difference between 0 h oxygen supply group and the control group.The expression of PPARγ was decreased both at mRNA and protein level after 6 h of oxygen supply. The difference between 6 h oxygen supply group and the control group was significant(P<0.01), which decreased with the length of reperfusion and the lowest was at 48 h after the reperfusion. The difference among the different reperfusion groups and the control group was significant(P<0.01). Conclusion PPARγ may participate in the pathological damage course of neuron ischemical reperfusion injury, and may become a new intervention target of treatment for ischemic cerebrovascular disease.