Na B Activates Katp Channels To Regulate The Mechanism Of Pd Microglia Model Inflammation

Objective:By observing the effect of sodium butyrate（Na B）on the inflammation of microglia in Parkinson’s inflammatory cell model induced by MPP⁺（1-methyl-4-phenylpyridine）,to investigate whether the mechanism of sodiumbutyrate in regulating inflammation is related to ATP-sensitive potassium channel（KATP）.Methods:1.Use MPP⁺to prepare a Parkinson’s inflammatory cell model on BV2 microglia.The CCK8 method was used to detect the effects of different concentrations of MPP+and sodium butyrate on the activity of BV2 microglia after 24 hours,and select theappropriate concentration.2.Observe the morphological changes of BV2 cells in the blank control group and MPP⁺building block under an optical microscope.Cellular immunofluorescence technique was used to detect the expression of ionized calcium binding adapter molecule 1（Iba-1）on blank control group,MPP⁺module,Na B pretreatment group and pinacidil pretreatment group（positive control）3.A NO kit was used to compare the amount of NO produced by BV2 cells among five different groups.4.Detect the m RNA expression levels of inflammatory factors IL-1β,IL-6 and TNF-αin five groups by real-time fluorescence quantitative（q PCR）method,and observe the effect of sodium butyrate on the release of inflammatory factors.5.Colorimetric method was used to detect the changes of intracellular ATP content in blank control group,pinacidil pretreatment group（ie positive control group）,sodium butyrate group,MPP⁺module and sodium butyrate pretreatment group,and observe butyric acid The effect of sodium on the opening of K_ATPchannels on microglia.6.Using real-time fluorescence quantitative PCR（q PCR）and western blot methods to detect the expression of K_ATPchannel subunits in the blank control group,MPP⁺module and sodium butyrate pretreatment groupResults:1.The results of CCK8 show that after treatment of BV2 microglia with different concentrations（50μmol/L-2mmol/L）of MPP⁺,it is divided from the blank area,and the average value of MPP⁺at various concentrations decreases the cell activity and has statistical significance（P<0.05）;different concentrations of sodium butyrate（1.25mmol/L-10mmol/L）have no toxic effect on BV2 microglia.Choose 100μmol/L MPP⁺for experiment.2.Divide BV2 microglia into 4 groups,namely blank control group,MPP⁺model,Na B pretreatment group and pinacidil pretreatment group（ie positive control group）.3.Observe the morphological changes of BV2 cells in the blank control group and MPP⁺module under the light microscope.Cell immunofluorescence was used to detect the expression of ionized calcium binding adapter molecule 1（Iba-1）on BV2microglia cells in 4 groups.3.Use NO kit to compare the amount of NO produced by BV2 cells in different groups.4.Detect the m RNA expression levels of inflammatory factors IL-1β,IL-6 and TNF-αin the four groups by real-time fluorescence quantitative（q PCR）method,and observe the effect of sodium butyrate on the release of inflammatory factors.5.Colorimetric method was used to detect the changes of ATP content in the four groups of cells,and to observe the effect of sodium butyrate on the opening of K_ATPchannels on microglia.6.Compared with the blank control group,the ATP content in BV2 microglia in the MPP⁺model was significantly reduced,and like the pinacilil pretreatment group,the sodium butyrate pretreatment group reversed the BV2 microglia The decreased ATP content in the cells indicates that sodium butyrate promotes the opening of K_ATPchannels on BV2 microglia.Conclusion:1.Sodium butyrate has a protective effect on MPP⁺activated BV2 microglia.2.Sodium butyrate can inhibit the activation of BV2 microglia induced by MPP+and inhibit the release of NO and inflammatory factors.3.Activation of K_ATPchannels can also inhibit MPP⁺-induced activation of BV2microglia,reducing the production of NO and inflammatory factors.4.In the PD inflammatory cell model,sodium butyrate regulates K_ATPchannels byup-regulating the expression of Kir6.1 and Kir6.2 subunits and promoting the opening of K_ATPchannels,thereby inhibiting the activation of microglia and reducing The NO and pro-inflammatory factors released by microglia reduce the inflammatoryresponse.