Study On The Common Mechanism Of The Method Of Tonifying The Kidney And Replenishing Essence In The Treatment Of Primary Osteoporosis And Alzheimer's Disease

Objective: 1.To compare the BMD between AD patients and non-AD controls.2.To explore the common disease-related genes between OP and AD patients,and to verify the common gene and its signaling pathway with OP and AD patients serum samples and animal models.3.To observe the effects of kidney tonifying formula on OP and AD mice models and its regulation of HIPK1 and its signaling pathway.Methods: 1.Cross-sectional studies,cohort studies or epidemiological investigations which contain BMD values of AD patients and non-AD controls by DXA,were searched through Pub Med,Clinical Trail,CNKI,Wan Fang,Sino Med and VIP.Information including title,author,publication year,state,study type,sample size,age,BMI,diagnosis of AD,BMD position and BMD were extracted.NOS is adopted to evaluate the quality of the researches,Rev Man 5.3 is used for statistical analyses.2.Chip information of OP and AD patients and their controls in NCBI GEO Data Sets were extracted.Differential gene expression of postmenopausal women was screened via comparison between high BMD and low BMD groups,or via comparison between AD.HI and non AD.HI.Moreover,gene enrichment analysis was made via KEGG pathway analysis.Blood serum of OP patients,AD patients and control women were collected to evaluate the expression level of TNF-α,and the expression patterns of its downstream molecules including HIPK1 and SENP1 were detected with AD and OVX mice model.Brain tissues and lumbar vertebra of 6-month old OVX mice,AD mice and their control mice were collected;IHC staining was performed to compare the protein expressions of HIPK1 and SENP1.3.3-month old APP/PS1 mice were divided into AD Model group,AD FF group,AD ALS group,AD+OP Model group,AD+OP FF group and AD+OP YX group.Wild-type mice were divided into AD Control group,OP Model group,OP Sham group,OP FF group and OP FSM group.Mice of OP Sham group,AD Model group,AD Control group and AD+OP Model group were intragastrically administrated with phosphate buffered saline;mice of OP FF group,AD FF group and AD+OP FF group were intragastrically administrated with kidney nourishing formula;mice of OP FSM group were intragastrically administrated with alendronate;mice of AD ALS group were intragastrically administrated with donepezil;mice of AD+OP YX group were intragastrically administrated with alendronate and donepezil together.Three months later,escape latency and numbers crossing platform were calculated through Morris water maze;bone mineral density of lumbar 5 vertebrae was scanned through Micro-CT;morphology of bone L5 vertebrae was observed through HE staining,and hippocampus area through Nissl staining;total protein expressions of HIPK1 and SENP1,expression of HIPK1 in cytoplasm and expression of SENP1 in nucleus were calculated.Results: 1.Compared with non-AD controls,AD patients’ whole body BMD were lower,MD(95%CI)is-1.32(-2.48,-0.16),the difference is statistically significant(P=0.03).Subgroup analysis also shows significant difference in femoral neck BMD between AD patients’ and non-AD,MD(95%CI)is-0.17(-0.18,-0.15),the difference is statistically significant(P<0.01).2.(1)Hipk1,Brsk2,Pcsk1 n,Phactr1,Rcan2 and Slc25a14 are the common genes in both OP and AD,of which Hipk1 is commonly up-regulated.(2)TNF-α in AD patients was higher than Sham group[(9.59±2.77)pg/m L vs(5.79±1.77)pg/m L,P=0.01],TNF-α in OP patients was also more than Sham group[(6.93±2.57)pg/m L vs(5.79±1.77)pg/m L,P=0.36],there was no significant difference in TNF-α between OP and AD patients(P=0.09).(3)Total protein expression of HIPK1 in hippocampus of AD Model group were more than Sham group(P<0.01).Total protein expression of HIPK1 in lumbar of OP Model was more than Sham group(P=0.006).Total protein expression of SENP1 in hippocampus of AD Model was more than Sham group(P<0.01).Total protein expression of SENP1 in lumbar of OP Model was more than Sham group(P=0.013).Expression of SENP1 in nucleus in hippocampus of AD Model was more than Sham group(P=0.002).Expression of SENP1 in nucleus in lumbar of OP Model was more than Sham group(P<0.01).Expression of HIPK1 in cytoplasm in hippocampus of AD Model was more than Sham group(P=0.002).Expression of HIPK1 in cytoplasm in lumbar of OP Model was more than Sham group(P<0.01).3.(1)Escape latency of AD ALS group and AD FF group were shorter than AD Model group(both P<0.01).Escape latency is significantly different among AD+OP YX,AD+OP FF and AD+OP Model group,however no difference among OP groups.Numbers crossing platform of AD Model and AD FF group are less than AD Control group(both P<0.05).(2)Discontinuitous,bone trabecula in OP Model,AD+OP Model is fewer and fractured,bone medullary cavity is bigger.Irregular arranged,cellular degeneration and necrosis were found in hippocampus of AD Model and AD+OP Model.Positive medicine and formula can improve the tissue formation and bone mass,BV/TV,Tb.N,Tb.Th in OP Model,AD+OP Model group is declined,while Tb.Sp lifted.(3)Total protein expression of HIPK1 and SENP1 were up-regulated in OP Model,AD Model,AD+OP Model in hippocampus and lumbar vertebra.HIPK1 is significantly up-regulatd in cytoplasm while SENP1 up-regulated in nucleus.Positive medicine and formula can also improve the total protein expression of HIPK1 and SENP1 in hippocampus and lumbar vertebra,expression of HIPK1 in cytoplasm and expression of SENP1 in nucleus.Conclusions: 1.AD patients had lower whole body and femoral neck BMD compared with non-AD controls.2.Hipk1 is one of the common up-regulated gene in both OP and AD patients,its upstream TNF-α is up-regulated in the serum of OP and AD patients.Total protein expressions of HIPK1 and SENP1,as well as intranuclear SENP1 expression and intracytoplasmic HIPK1 expression,in hippocampus and lumbar vertebrae were up-regulated.3.Bone mass in OVX mice and cognitive function in AD mice could be improved by the treatment of kidney nourishing formula.Total protein expression of HIPK1 and SENP1 in hippocampus and lumbar vertebra,as well as intranuclear SENP1 expression and intracytoplasmic HIPK1 expression,were reduced by kidney nourishing formula.Kidney nourishing formula might improve OP and AD mice through regulation of TNF-α-SENP1-HIPK1 pathway.