| Objective: This study aims to explore new biomarkers and candidate therapeutic targets for Alzheimer’s disease by exploring whether dexmedetomidine can cause the apoptosis of neurons in Alzheimer’s disease by down-regulating Lnc RNA SNHG14.Method:1.Construct an in vitro cell model of Alzheimer’s disease,use Aβ1-42 to treat SH-SY5 Y cells to simulate the in vitro cell model of AD,detect cell viability by CCK8 assay,and detect the expression level of Lnc RNA SNHG14 by qPCR,so as to determine the AD cell model Whether the build is successful or not.2.Screen the appropriate concentration of dexmedetomidine,treat SH-SY5 Y cells with different concentrations(0,5,10 and 50 μM)of dexmedetomidine,and detect cell viability by CCK8 method to determine the efficacy of dexmedetomidine.optimal intervention concentration.3.To observe the inhibition of dexmedetomidine on the apoptosis of AD model cells,divide the experimental cells into three groups: Control group(control group),Aβ group(AD model cell group),Dex group(dexmedetomidine treatment group).The cell viability of the experimental samples was detected by the CCK8 method,the gene expression of SNHG14 was detected by the qPCR method,the apoptosis of the experimental cells was detected by flow cytometry,and the expression of apoptosis-related proteins was detected by Western Blot technology.4.To explore whether dexmedetomidine can play an anti-apoptotic role in AD cells by regulating the expression of Lnc RNA SNHG14.The experimental grouping is the same as above.The overexpression cell model of SNHG14 is constructed by transfecting oe-SNHG14,and whether SNHG14 is detected by qPCR technology.The transfection was successful,and the cell viability of the experimental samples was detected by the CCK8 method,the apoptosis of the experimental cells was detected by flow cytometry,and the expression of apoptosisrelated proteins was detected by Western Blot technology.Result:1.The AD cell model was successfully constructed,and the qPCR detection results of the dexmedetomidine-acting gene ADRA2 showed that there was no statistical difference in the expression of ADRA2 in SH-SY5 Y cells treated with different concentrations of Aβ1-42(P>0.05);CCK8 The experimental results showed that the cell viability of SH-SY5 Y cells treated with different concentrations of Aβ1-42 decreased sequentially(P<0.05),and the qPCR detection results of SNHG14 gene showed that the expression of SNHG14 was up-regulated in a concentration-dependent manner with Aβ1-42(P<0.05).2.Successfully screened out the appropriate concentration of dexmedetomidine to treat model cells,treated SH-SY5 Y tool cells with different concentrations of dexmedetomidine,and detected its cell activity by CCK8 method.The results showed that when exposed to At 50 μM Dex,the viability of SH-SY5 Y cells was dead,but there was no significant difference in cell viability after treatment with other concentrations(P>0.05),so we chose a relatively high concentration(10 μM)of dexmedetomidine for subsequent experiments.3.Dexmedetomidine can inhibit the apoptosis of AD model cells.The results of qPCR showed that compared with the control group,the expression of Lnc RNA SNHG14 in the cells of the Aβ group increased(P<0.001),and the expression of Lnc RNA SNHG14 in the cells of the Dex group Decreased(P<0.05);after adding DEX blocker,the expression of Lnc RNA SNHG14 in Aβ group and Dex group cells was significantly decreased compared with that before treatment(P<0.01).The results of CCK8 detection showed that compared with the control group,the cell viability of the Aβ group decreased,while the cell viability of the Dex group was significantly enhanced(P<0.05).The results of Annexin V-FITC showed that compared with the control group,the apoptosis of cells in Aβ group was significantly increased(P<0.001);the apoptosis of cells in Dex group was significantly decreased(P<0.01).The results of Western Blot showed that compared with the control group,the apoptosis-related protein Bax in the Aβ group was significantly increased,Bcl2 was significantly decreased,and the expression level of p-tau/tau,an important biomarker for AD diagnosis,was increased in the Aβgroup;while the Dex group could significantly antagonize the above-mentioned phenomenon(P<0.05).4.The overexpression of SNHG14 can promote the apoptosis of AD model cells.The results of qPCR examination showed that the expression of Lnc RNA SNHG14 in the Aβ+oeSNHG14 group was significantly increased compared with the Aβ group(P<0.001).The results of CCK8 showed that compared with the Dex group,the cell viability of the Aβ+Dex+oeSNHG14 group decreased(P<0.05).The results of Annexin V-FITC showed that compared with the Dex group,the apoptosis of cells in the Aβ+Dex+oe-SNHG14 group was significantly increased(P<0.05).The results of Western Blot showed that compared with the Dex group,the apoptosis-related protein Bax in the Aβ+Dex+oe-SNHG14 group was significantly increased,Bcl2 was significantly decreased,and the expression level of p-tau/tau,an important biomarker for AD diagnosis,was increased(P< 0.05).Conclusion: Dex can inhibit neuronal apoptosis in AD model by down-regulating the level of lnc RNA SNHG14. |
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