| Part ? Detection of serum miR-182 in patients with Alzheimer's disease and its clinical significance ObjectiveTo investigate the changes of miR-182 concentration in serum of patients with Alzheimer's disease(AD)and their clinical significances.MethodsCollected serum samples from 40 patients with first-time AD before and after memantine treatment.Forty healthy volunteers were collected as control group.The concentration of serum miR-182 was detected by realtime fluorescence quantitative PCR.ROC curve was used to evaluate the sensitivity and specificity of serum miR-182 expression level to the diagnosis of AD.At the same time,the scores of mini-mental state examination(MMSE)before and after treatment were evaluated in the patients with AD,and the correlation between the relative expression level of serum miR-182 and the scores of MMSE was analyzed.Results1.Serum miR-182 expression in patients with Alzheimer's disease was significantly higher than in the control group(z=-7.612,P<0.01).2.After 4 months of memantine treatment,the expression of miR-182 in the serum of AD patients was significantly lower than that before treatment(z=-4.302,P<0.05).3.There was a negative correlation between serum miR-182 expression level and MMSE score(r =-0.546,P<0.05).4.The area under the serum miR-182 ROC curve for AD diagnosis was 0.904(95%CI: 0.846~0.963,P<0.01),the sensitivity was 89.1%,and the specificity was 80.0%.ConclusionThis study shows that miR-182 can be used as a serum marker of AD and has good clinical application value in the diagnosis and evaluation of AD.Part ? Effects of mi R-182 on cell proliferation and BDNF expression in AD model cellsObjective By verifying the expression of mi R-182 in AD model cells,the effect of mi R-182 on the proliferation of AD model cells was discussed,and the relationship between mi R-182 and target genes was further explored,so as to preliminary explore the mechanism of mi R-182 in AD.Methods SH-SY5 Y cells,which cultured in vitro,were induced by different concentrations of A?1-40 to construct cell models.q PCR was used to detect mi R-182 expression in normal SH-SY5 Y cells and model cells.Lipofectamine2000 transient transfection method was used to transfect AD model cells,and the effect of transfection was observed by q PCR experiment.CCK-8 test was used to detect the effect of mi R-182 on the proliferation activity of AD model cells.The potential downstream target gene BDNF of mi R-182 was predicted by two bioinformatics databases,Miranda and targetscan.The effects of overexpression and knockdown of mi R-182 on BDNF m RNA and protein levels were detected by q PCR and Western blot.Results 1.The expression level of mi R-182 in AD model cells was significantly higher than that in normal SH-SY5 Y cells(P<0.05).2.Compared with the control group,the transfection of mi R-182 can significantly inhibit cell proliferation,while the si RNA-mi R-182 can significantly increase cell proliferation(P<0.05).3.The prediction results of online database show that there is a potential complementary base binding site between mi R-182 and 3'UTR of BDNF.4.Overexpression of mi R-182 significantly inhibited the expression of BDNF mRNA and protein,while knockdown of mi R-182 significantly increased the expression of BDNF m RNA and protein(P<0.05).Conclusion This study suggests that miR-182 can play a role in inhibiting cell proliferation and reducing BDNF expression in the pathogenesis of AD,providing new experimental support for further revealing the pathogenesis of AD. |
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