| Alzheimer's disease(AD)is a progressive neurodegenerative disease with an increasing incidence,which brings tremendous pressure and challenges to AD patients' families and human society.The curative effect of modern medicine on AD is not ideal.Traditional Chinese medicine(TCM)has a long history of understanding dementia.With its multi-target,multi-channel,and synergistic characteristics,Chinese herbal medicine compounds can play an overall therapeutic effect on AD.Guilingji Capsule(GLJC),as a famous prescription for invigorating the kidney and anti-aging,has the functions of invigorating the kidney qi,strengthening the body,and replenishing the brain.Exploring its therapeutic effects and potential mechanisms for AD has important value to the prevention and treatment of AD with TCM.Objective:To evaluate the clinical efficacy of GLJC on the treatment of patients with AD,bioinformatics analysis was used to explore the potential mechanism of GLJC,thus promoting the clinical application of GLJC in AD therapy.Methods:1 Clinical research:we conducted a double-blind,double-simulation randomized controlled trial(RCT).60 AD patients with kidney deficiency and encephala reduction were enrolled and divided into experimental groups and control groups.The experimental group was treated with GLJC+Ginkgo biloba simulator,and the control group was treated with Ginkgo biloba+GLJC simulator.The total course of treatment was 24 weeks.At the baseline,midpoint,and endpoint,the changes of MMSE,ADAS-cog,ADL,and TCM syndrome were observed,as well as the safety of intervention drugs was evaluated.2 Screening and verification of bioinformatics analysis:All microarray datasets were obtained from the GEO database.DEGs were screened from the GSE132903 dataset that compared the gene expression of the middle temporal gyrus in AD patients and non-demented controls.These DEGs were further analyzed by WGCNA to obtain AD-related DEGs.GO and KEGG pathway analyses were conducted utilizing the DAVID tool.Whilst we constructed a PPI network through the STRING and used Cytoscape software to extract hub genes.DEGs in the GSE63060 dataset whose sample sources were blood tissues were intersected with hub genes screened from the brain tissues to screen out hub genes that were co-expressed in the brain and blood of AD patients.AD group and non-AD control group were compared to verify the expression of hub genes in the blood.The effect of GLJC on the expression of these hub genes was detected.Results:1 Results of clinical research:(1)Primary outcomes:compared with baseline,the MMSE scores of the experimental group and control group were significantly increased at the 12th weeks and 24th weeks(P<0.01).At the 12th weeks and 24th weeks,there was no statistical difference in MMSE scores among groups(P>0.05).Compared with baseline,the ADAS-cog scores of the experimental group and control group were significantly declined at the 12th weeks and 24th weeks(P<0.01).At the 12th weeks and 24th weeks,there was no statistical difference in ADAS-cog scores among groups(P>0.05).(2)Secondary outcomes:compared with baseline,the TCM syndrome scores of the experimental group and control group were significantly declined at the 12th weeks and 24th weeks(P<0.01).At the 12th weeks and 24th weeks,there was no statistical difference in TCM syndrome scores among groups(P>0.05).Compared with baseline,the ADL scores of the experimental group and control group were significantly declined at the 12th weeks and 24th weeks(P<0.01).At the 12th weeks and 24th weeks,there was no statistical difference in ADL scores among groups(P>0.05).(3)Safety evaluation:during the treatment period,the two groups of patients had no adverse reactions,and there were no abnormal changes in blood routine,urine routine,stool routine,liver function,renal function,and electrocardiogram.2 Results of bioinformatics analysis:755 DEGs were screened from GSE132903.The WGCNA of these DEGs showed that the 364 DEGs gathered in turquoise modules were significantly negatively correlated with AD.At biological process,these 364 DEGs significantly related to AD mainly involved the establishment of organelle localization,signal release,and synaptic organization,etc.At cellular components,DEGs mainly involved presynapse,glutamatergic synapses,cell leading edge,etc.At molecular function,DEGs mainly involved calmodulin binding,channel regulator activity,ion channel regulator activity,etc.The biological pathways that DEGs participate in mainly involved calcium signaling pathways,cGMP-PKG signaling pathways,GABAergic synapses,etc.The top 20 hub genes with the highest degree of enrichment in PPI network analysis:STY1,ITSN1,AMPH,DNM3,CLTA,NECAP1,REPS2,GABRG2,GFAP,GABRA1,BDNF,ITPR1,ITPR3,ATP2A2,PPP3CB,NRXN1,GAD1,SST,NPY,PPP3CA.The hub genes that are co-expressed in the brain and blood of AD patients include PPP3CB and ITPR3.3 RT-PCR verification and results after GLJC intervention:Compared with the non-AD control group,PPP3CB was significantly lower expressed in the blood of AD patients(P<0.01).There was no significant difference in the expression of ITPR3 in the blood between the non-AD control group and the AD group(P>0.05).After the intervention of GLJC,the expression of PPP3CB in the blood of AD group was significantly increased,with statistical difference(P<0.05).Conclusions:1 Conclusion of clinical research:GLJC can increase the MMSE score of AD patients with kidney deficiency and encephala reduction,reduce ADAS-cog,ADL scores,and TCM syndrome scores,and improve the cognitive function and ability of daily living in AD patients.During the treatment,there were no adverse reactions and the safety was good.2 Conclusion of bioinformatics analysis and RT-PCR verification:PPP3CB was significantly down-regulated in the blood of AD patients.GLJC can significantly up-regulate the expression of PPP3CB in AD patients.GLJC may exert a therapeutic effect on AD by regulating PPP3CB. |
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