| Alzheimer’s disease(Alzheimer’s disease,AD)is a kind of nervous system degenerative disease that closely related to age.AD is characterized by memory loss and cognitive impairment in clinical feature and intracerebral amyloid(β-amyloid,Aβ)deposition,neural fiber entanglement formation and neuron loss in pathological performance,respectively.AD includes two kinds of cases,familial AD and sporadic AD.Studies show that the pathological mechanism may be associated with a number of factors,such as Aβ deposition,ion channel,calcium metabolism disorder,vascular anomalies,abnormal tau protein,oxidative stress and so on.With the deepening of the research,we find that Aβ plays an important role in the process of AD.Due to the abnormal metabolism,the balance between generation to remove of Aβ was broken,triggering Aβ deposition.A variety of enzymes played a significant role in this dynamic process,including β-,γ-secretase related to generating and the NEP,IDE,MMP-2/9 related to degradation,etc.,which maintain the balance in the content of Aβ together.Until now,we still don’t know clearly whether lower testosterone level is a reason or a result in AD.Some studies suggest that the decrease in testosterone may as a risk factor for AD,which occur in the early stage of AD or before AD.A research of longitudinal prospective study on aging compared the diagnosis with the change of blood testosterone levels in AD.The men who were normal in the first clinical diagnosis were followed up for 4 to 37years(an average of 19 years),and the final results showed testosterone level dropped and that decline occurred 5 to 10 years before the diagnosis of AD,suggestting that testosterone decline in AD before clinical symptoms.Rosario and his colleagues found testosterone levels decline with age from 50 to 97-year-old male midbrain organization by autopsy,moreover,the extent of the decline in patients with AD is significantly higher than normal.Compared with the patients who suffered in early stage of AD,the patients who had mild nerve pathological changes with brain tissue had lower testosterone levels,suggestting that testosterone decline occurred before the pathological changes and may promote the progress of AD.Age-related testosterone decline is a risk factor for AD.Androgens play a role in AD development mainly through the following three ways:(1)Androgen receptor(Androgen receptor,AR)pathways.There are androgen receptor in the hypothalamus,hippocampus and temporal lobe cortex and the testosterone can be tansformed into biological efficacy dihydrotestosterone under the action of 5α-reductase.Both testosterone and dihydrotestosterone play a role through combine with AR.(2)Estrogen receptor(Estrogen receptor,ER)pathways.As a kind of prohormone,testosterone can be transformed into 17β-estradiol under the action of aromatase and the latter influence indirectly the occurrence of AD development through combine with ER.(3)Gonadotropin pathways.Testosterone is one of the important hormones in the hypothalamus – pituitary-gonadal axis,whose decreasing level could affect other hormones’ level by negative feedbacks.The results that luteinizing hormone and follicle-stimulating hormone decline may be associated with AD.There are some possible mechanism of androgen in the development of AD,such as adjusting Aβ gathered,reducing tau phosphorylation,increasing the synaptic plasticity,reducing oxidative stress,adjusting the mitogen activated protein kinase/extracellular signal regulating kinase signaling pathways,increase the expression of nerve growth factor and inhibiting the inflammatory response.A follow-up find from androgen deprivation therapy for prostate cancer patients who were in the mild nervous pathological changes of AD revealed that the level of serum testosterone and estradiol declined sharply,however the level of Aβ oviously increased,which showed that the level of brain tissue Aβ was inversely proportional to the level of testosterone.As previously mentioned,androgen adjust Aβ level through direct AR pathways or indirect ER pathway.Testosterone can adjust the Aβ level eather by reduce Aβ production or by increase the Aβ remove.There are two main competing cracking way for Aβ precursor(Amyloid precursor protein,APP): APP transformes into Aβ thought the action of β-and γ-enzymes,the other way dose not produce amyloid flexible through α-secretase,which prevents the generation of Aβ.Testosterone adjusts the the APP originating from cerebral cortex neurons and reduces the generation of Aβ by increases non-amyloidosis APP fragment secretion.Aβ can be degraded into small molecular peptide under the role of some protein enzymes and cleared from the body,while the dysfunction of these enzymes contribute to the pathogenesis of AD.The present study shows that there are some main degradation enzymes as followed,NEP,IDE and MMPs.The decline of androgen leaded to the decrease level of NEP and the increase level of Aβ,which could be recovered by DHT replacement therapy,suggestting that androgen play a role in regulation of Aβ by NEP.Androgen may also adjust Aβ by the thyroid hormone carrier protein.Extracellular matrix metalloproteinases inducing factor(Extracellular matrix metalloproteinase inducer,CD147)is a kind of type I transmembrane glycoprotein,belonging to the immunoglobulin superfamily,stimulates tumor cells secrete matrix metalloproteinase and fibroblasts in the metal protein enzymes(MMPs),leading to degradate basement membrane and stroma cells components,which play an important role in the process of tumor invasion and metastasis in.But in recent years,studies have reported that CD147 has related to the production of Aβ and mediation in AD,but its specific molecular mechanism is still not very clear.The research shows that CD147 is another subunit of γ-secretase complexs,participate in the negative regulation βamyloid precursor protein(APP)to produce Aβ,and participate in the remove of Aβ.Part one The effect of dihydrotestosterone on the expression of extracellular matrix metalloproteinase inducer in SH-SY5Y cellsObjective: Take SH-SY5Y cells as the research object to observe the effect of DHT on the expression of CD147 by immunofluorescence and Western blot.Whether flutamide,a classic androgen nuclear receptor inhibitor,could block the increasing level of CD147 protein mediated by DHT or not.Further using Chip technology to prove the existence of CD147 gene promoter region where AR could bind.Methods:1.Dose-effect experiment of DHT on the expression of CD147 in SH-SY5Y cellsExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(0 h),1 n M treatment group(1 n M),10 n M treatment group(10 n M),100 n M treatment group(100 n M)and control group was treated with the same amount of DMSO.The administration time was 6 h.After disposure,protein was extracted and performed Western blot to detect the protein changes of CD147 by different dose DHT.2.The effect of pretreatment flutamide on the expression of CD147 by immunofluorescence cytochemistryExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(Con),flutamide+DHT group(F+DHT),dihydrotestosterone(DHT).According to the result of the first part of the experiment,the DHT administration time and dose in F+DHT and DHT groups were 6 h and 100 n M,respectively.Flutamide pretreated for 1 h.After disposure,immunofluorescence cytochemical staining of CD147 was conducted,for the purpose of observing the alternation of CD147 by DHT and flutamide.3.The effect of pretreatment flutamide on the expression of CD147 by Western blotExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(Con),flutamide+DHT group(F+DHT),dihydrotestosterone(DHT).According to the result of the first part of the experiment,the DHT administration time and dose in F+DHT and DHT groups were 6 h and 100 n M,respectively.Flutamide pretreated for 1 h.After disposure,protein was extracted and performed Western blot to detect the relative expression level of CD147 in each group.4.To detect the androgen receptor binding site of CD147 promoter region by oprecipitation of chromatin experimentsExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into blank group(Blank),Ig G group(Ig G),Inpu group(Input)and AR group(AR).Put AR specificity antibody to AR group,set homology Ig G to Ig G group,put positive control antibody provided by kit to Input group kit,join sterile water to the Blank groupe.Through crosslinking,cracking,ultrasonic treatment fracture of DNA,crosslinked protein/DNA precipitation,protein/DNA complexes of elution,protein/DNA complexes with free DNA against crosslinking,purification,Realtime-PCR,agarose gel electrophoresis to detect CD147 promoter region exists androgen receptor binding site.The products were performed sequencing.Results:1.Dose-effect experiment of DHT on the expression of CD147 in SH-SY5Y cellsApplied to different doses of DHT to SH-SY5Y cells for 6 h,relative expression values of CD147 were respectively 0.26±0.02(0 n M),0.27±0.01(1 n M),0.31±0.01(10 n M),0.35±0.02(100 n M).The data of 100 n M group were significantly higher than other three groups(P<0.05).The data of 10 n M group was increased to some extent compared with 1 n M and 0 n M group(P<0.05).But there was no significant difference between 0 n M and 1 n M group(P>0.05).2.The effect of pretreatment flutamide on the expression of CD147 The immunofluorescence staining results showed that: compared with Con group(0.046±0.007),the average of fluorescence intensity increased slightly in F+DHT group(0.054±0.003),while increased significantly in DHT group(0.069±0.005)(P<0.05).Compared with DHT group(0.069±0.005),F+DHT group(0.054±0.003)fluorescence intensity was reduced(P <0.05).3.The effect of pretreatment flutamide on the expression of CD147 The Western blot results showed that: compared with Con group(0.54±0.02),the protein level of CD147 in F+DHT group(0.60±0.04)was increased and also in DHT group(0.67 ± 0.04)was obviously increased(P<0.05).Compared with DHT group(0.67 ± 0.04),the protein level of CD147 in F+DHT(0.60±0.04)group was reduced(P<0.05).Each band is relative to the internal optical density value of GAPDH.4.To detect the androgen receptor binding site of CD147 promoter region The scope of the DNA fragments size mainly presented within 200 to1000 bp,which could be used for follow-up study.Real-time PCR results showed that: the PCR amplification peak appeared earliest in Input group(24.93±0.09),followed by AR group(27.32±0.07),Ig G group(30.51±0.24)and Blank group(43.13±4.00).Agarose gel electrophoresis showed that: the banding of Input group was brightest and the IOD value was 50645±7550.64,followed by AR group 27042 ± 7683.61 and Ig G group 13692 ±1886.75,but Blank group did not show the stripe.Amplification products by sequencing test analysis were consistent with expected base sequence AAGGACGTTCTGACC.Part two The role of extracellular matrix metalloproteinase inducer on regulation of the cellular damage induced by Aβ42 in SH-SY5Y cellsObjective: Take SH-SY5Y cells as the research object to observ the changes of cell vitality,morphology,apoptosis related proteins and Aβ42 in the cell culture medium supernatant,following by interfered with LV-BSG-RNAi lentivirus and exogenous Aβ cell damage.To investigate the relationship among CD147,Aβ42 clearance and cell damage.Methods:1.The effect of Aβ42 on the cell vitality in SH-SY5Y cells Experiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(0 h),1 μM Aβ42 treatment group(1μM),10 μM Aβ42 treatment group(10 μM),20 μM treatment group(20 μM)and 40 μM Aβ42 treatment group(40 μM).The cells were cultured in 100 μl serum free medium containing different Aβ42 concentrations and the control group culture contained the amount of PBS,with each concentration included three parallel holes.After disposure 24 h,each hole was added MTS kit reagent 20 μl and then incubated for 2 h in 37 ℃.Absorbance value were obtained by Infinite200 enzyme standard meter in 490 nm in order to observe the influence of Aβ42 on the cell vitality.2.The detection of infection efficiency for lentivirus LV-BSG-RNAi in SH-SY5Y cellsExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(Con)and interference group(RNAi).Control group and RNAi groups were added negative control slow virus Con077(1E+9 TU/ml)and interference LV-BSG-RNAi lentivirus(8E+8TU/ml),respectively.After disposure 12 h,the culture was replaced by conventional culture medium and then incubated for 72 h in the condition of37℃ 5% CO2 incubator.Efficiency of infection was observed by inverted microscope and tested by Western blot and RT-PCR methords.3.The role of interference CD147 expression for extracellular Aβ42 clear in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+Aβ42 group(Con+Aβ42),interference(RNAi),interference+Aβ42 group(RNAi+Aβ42).Based on the results of part one,we administrated Aβ42 of 10 μM for 24 h and then collected the cell supernatant to test the changes of Aβ42 by Elisa.4.The role of interference CD147 expression for cell viability SH-SY5YExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+Aβ42 group(Con+Aβ42),interference(RNAi),interference+Aβ42 group(RNAi+Aβ42).The cells were cultured in 100 μl serum free medium containing 10 μM Aβ42.After disposure24 h,each hole was added MTS kit reagent 20 μl and then incubated 2 h in 37℃.Absorbance value were obtained by Infinite200 enzyme standard meter in 490 nm in order to observe the cell vitality changes in different groups.5.The role of interference CD147 expression for cell morphology in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+Aβ42 group(Con+Aβ42),interference(RNAi),interference+Aβ42 group(RNAi+Aβ42).The cells were incubated with 10 μM Aβ42 for 24 h and then were observed by inverted microscope for the cellular morphological changes.6.The role of interference CD147 expression for cellular nucleus morphology induced by Aβ42 in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+Aβ42 group(Con+Aβ42),interference(RNAi),interference+Aβ42 group(RNAi+Aβ42).The cells were incubated with 10 μM Aβ42 for 24 h and then were performed Hoechst 33258.The cells were observed by inverted microscope for the cellular nucleus morphological changes.7.The role of interference CD147 expression for the regulation of apoptosis related proteins due to Aβ42 in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+Aβ42 group(Con+Aβ42),interference(RNAi),interference+Aβ42 group(RNAi+Aβ42).The cells were incubated with 10 μM Aβ42 for 24 h and then were collected to detect apoptosis related proteins Bax and Bcl-2 by Western blot methord.Results:1.The effect of Aβ42 on the cell vitality in SH-SY5Y cells According to the results of MTS test,the cellular vitality was decrease followed by incubated for 24 h 10 μM Aβ42.Compared with the control group(0.80±0.04),1 μM group(0.75±0.01)did not change obviously(P>0.05).The rest of other groups were 10 μM 0.65±0.01,20 μM group 0.59±0.01 and 40μM group 0.48±0.01,respectively(P<0.05).Subsequent experiments were selected 10 μM Aβ42 incubated 24 h for test.2.The detection of infection efficiency for lentivirus LV-BSG-RNAi in SH-SY5Y cellsInfection rate can reach more than 80% after infected for 3 days.RT-PCR test results showed that: compared with Con group,CD147 m RNA level decreased by 65.60±4.26% in RNAi group(P<0.01);Western blot test results showed that: compared with Con group,CD147 the protein levels decreased by 73.62±6.31% in RNAi group(P<0.01).Subsequent experiments were selected the cells infected the lentivirus after 3 days.3.The role of interference CD147 expression for extracellular Aβ42 clear in SH-SY5Y cellsElisa results showed that: the Con(8.67±2.02 pg/ml)and RNAi(8.92±1.48 pg/ml)group were not added exogenous Aβ42 and the Aβ42 content in culture medium supernatant was lower Aβ42,with no obvious changes between the thess two groups(P>0.05).the Con+Aβ42(72.94±4.64)and RNAi+Aβ42(92.48±6.29)groups were added the exogenous Aβ42 and the Aβ42content in culture medium supernatant was higher(P>0.05).,Compared to Con+Aβ42 group(72.94±4.64),the Aβ42 content was obviously higher in RNAi+Aβ42 group(92.48±6.29),with statistically significant difference(P<0.05).4.The role of interference CD147 expression for cell vitality SH-SY5YMTS test results showed that: compared with Con group(0.77±0.05),the cell vitality declined obviously in Con+Aβ42 group(0.57±0.06)and RNAi+Aβ42 group(0.39±0.03)(P<0.01).But the RNAi group(0.72±0.03)had no obvious change(P>0.05).Compared with RNAi group(0.72±0.03),the cell vitality declined obviously in Con+Aβ42 group(0.57±0.06)and RNAi+Aβ42 group(0.39±0.03)(P<0.01).Compared with Con+Aβ42 group(0.57±0.06),the cell vitality declined obviously in RNAi+Aβ42 group(0.39±0.03)(P<0.01).5.The role of interference CD147 expression for cell morphology in SH-SY5Y cellsCells from Con group presented fusiform or triangular appearance with larger cell body and plump cytoplasm,mixed axons.Most cells from RNAi group presented fusiform or triangle appearance with some round soma and short axiaons.When incubated with 10 μM Aβ42 for 24 h,some of cell showed soma retraction,loss of refraction,emerge particle and axial dinimution in Con+Aβ42 group.While most of the cell showed soma retaction,aggregation,nearly circular or irregular shape,loss of refraction,short axial and even part cell fragments.6.The role of interference CD147 expression for cellular nucleus morphology induced by Aβ42 in SH-SY5Y cellsHoechst 33258 stainning showed that: nucleus were large,oval and uniformity dyeing in Con group;most of the nucleus had oval shape and uniformity dyeing in Con+Aβ42 except for some nucleus were pyknosis;most of the nucleus had oval shape and uniformity dyeing in RNAi group except for some nucleus were pyknosis and fragmentation;relative to the RNAi group,more pyknosis nucleus were present in RNAi+Aβ42 group.7.The role of interference CD147 expression for the regulation of apoptosis related proteins due to Aβ42 in SH-SY5Y cellsCD147 Western blot results show that: compared with Con group(0.82±0.05),the protein levels of CD147 in RNAi group(0.23±0.02)and RNAi+Aβ42 group(0.21±0.05)were significantly reduced(P<0.01).But there was no obviously change in Con+Aβ42 group(0.89±0.06)(P>0.05).There was no obviously change between RNAi group(0.23±0.02)and RNAi+Aβ42 group(0.21±0.05)(P>0.05).Compared with Con+Aβ42 group(0.89±0.06),the protein level of CD147 in RNAi+Aβ42 group(0.21±0.05)significantly reduced(P<0.01).Bax Western blot results showed that: compared with Con group(1.57±0.12),the protein levels of Bax enhanced in RNAi group(1.97 ± 0.34),Con+Aβ42 group(1.83 ± 0.19),RNAi+Aβ42 group(2.32 ± 0.20)(P<0.01).Compared with RNAi group(1.97±0.34),Con+Aβ42 group(1.83±0.19)had no obvious change(P>0.05),RNAi+Aβ42group(2.32 ± 0.20)increased significantly(P<0.01).Compared with Con+Aβ42group(1.83 ± 0.19),the level of Bax increased significantly in RNAi+Aβ42 group(2.32 ± 0.20)(P<0.01).Bcl-2 Western blot results showed that: compared with Con group(0.84±0.02),the Bcl-2 expression levels were lower in RNAi group(0.66±0.02),Con+Aβ42 group(0.64 ± 0.03),RNAi+Aβ42 group(0.33 ± 0.02)(P<0.01).Compared with RNAi group(0.66±0.02),Con+Aβ42 group(0.64±0.03)had no obvious change(P>0.05),RNAi+Aβ42 group(0.33 ± 0.02)significantly reduced(P<0.01).Compared with Con+Aβ42 group(0.64±0.03),the level of Bcl-2 reduced in RNAi+Aβ42 group(0.33±0.02)(P< 0.01).Part three Extracellular matrix metalloproteinase inducer takes part in the regulation of dihydrotestosterone against Aβ42 in damage progrese in SH-SY5Y cellsObjective: Take SH-SY5Y cells as the research object to observ the changes of cell vitality,morphology,apoptosis related proteins and Aβ42 in the cell culture medium supernatant,following by interfered with LV-BSG-RNAi lentivirus and exogenous Aβ cell damage with or withour pretreated DHT.To investigate the role of CD147 in the progress of DHT against the cell damage induced by Aβ42.Methods:1.The role of interference CD147 expression in the progress of DHT against the damage about cell vitality induced by Aβ42 in SH-SY5Y cellsExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(Con+Aβ42),interference group(RNAi+Aβ42),control group plus RNAi group(Con+DHT+Aβ42)and interference group plus DHT(RNAi+DHT+Aβ42).Control group and RNAi groups were added negative control lentivirus Con077(1E+9 TU/ml)or interference LV-BSG-RNAi lentivirus(8E+8 TU/ml),respectively.After disposure 12 h,the culture was replaced by conventional culture medium and then incubated for 72 h in the condition of 37℃ 5% CO2 incubator.After infected 3 days,the culture medium from Con+DHT+Aβ42 group and RNAi+DHT+Aβ42 group were replaced by serum free medium containing 100 nm DHT,while the culture medium from Con+Aβ42 group and RNAi+Aβ42group were replaced by serum free culture medium containing the same amount of DMSO.After exposed to 6 h,all culture medium was replaced by serum free medium containing 10 μM Aβ42 for 24 h and then cell vitality was detected in 490 nm.2.The role of interference CD147 expression in the progress of DHT adjust the clearance of extracelluar Aβ42 in SH-SY5Y cellsThe experimental group and drug delivery way were the same as experiment 1 Aβ42 content was detected by Elisa.3.The role of interference CD147 expression in the progress of DHT against the damage about cellular morphology induced by Aβ42 in SH-SY5Y cellsThe experimental group and drug delivery way were the same as experiment 1 cellular morphology was observed by microscope.4.The role of interference CD147 expression in the progress of DHT against the damage about cellular nucleus morphology induced by Aβ42 in SH-SY5Y cellsThe experimental group and drug delivery way were the same as experiment 1 cellular nucleus morphology was observed by microscope.5.The role of interference CD147 expression in the progress of DHT against the damage about Bax and Bcl-2 protein induced by Aβ42 in SH-SY5Y cellsThe experimental group and drug delivery way were the same as experiment 1 The protein levels were detected by Western blot.Results:1.The role of interference CD147 expression in the progress of DHT against the damage about cell vitality induced by Aβ42 in SH-SY5Y cellsMTS results showed that: compared with Con+Aβ42 compared group(0.49 ± 0.03),the leve of cell vitality of RNAi+Aβ42 group(0.34 ± 0.02)significantly decreased(P<0.05),but in Con+DHT+Aβ42group(0.63±0.03)and RNAi+DHT+Aβ42 group(0.56 ± 0.03)increased(P<0.05).Compared with RNAi+Aβ42 group(0.34 ± 0.02),the level of cell vitality of Con+DHT+Aβ42 group(0.63 ± 0.03)and RNAi+DHT+Aβ42 group(0.56 ±0.03)increased significantly(P<0.05).Compared with Con+DHT+Aβ42 group(0.63 ± 0.03),the cell vitality of RNAi+DHT+Aβ42 group(0.56 ± 0.03)reduced(P<0.05).2.The role of interference CD147 expression in the progress of DHT adjust the clearance of extracelluar Aβ42 in SH-SY5Y cellsElisa results showed that: compared with Con+Aβ42 group(65.40±4.16),the level of Aβ42 in RNAi+Aβ42 group(89.48±3.40)significantly increased(P<0.01),but the data in Con+DHT+Aβ42 group(35.37 ± 4.32)and RNAi+DHT+Aβ42 group(53.13 ± 2.34)significantly reduced(P<0.01).Compared to RNAi+Aβ42 group(89.48 ± 3.40),the level of Aβ42 in Con+DHT+Aβ42 group(35.37±4.32)and RNAi+DHT+Aβ42 group(53.13±2.34)significantly decreased(P<0.01).Compared with Con+DHT+Aβ42group(35.37±4.32),the level of Aβ42 in RNAi+DHT+Aβ42 group(53.13±2.34)increased significantly(P<0.01).3.The role of interference CD147 expression in the progress of DHT against the damage about cellular morphology induced by Aβ42 in SH-SY5Y cellsThe cells from Con+Aβ42 group showed cell body retraction,loss of refraction,emerged particles and thin axon;most of the cell from RNAi+Aβ42group presented body retraction,accumulation,nearly circular or irregular shape,loss of refraction,emerged particles,thin axon and even cell debris.Most cells from Con+DHT+Aβ42 group showed fusiform,triangular and some of them retract.Most cells from RNAi+DHT+Aβ42 group showed retraction and decrescent,nearly circular or irregular shape with thin axon.4.The role of interference CD147 expression in the progress of DHT against the damage about cellular nucleus morphology induced by Aβ42 in SH-SY5Y cellsHoechst33258 stainning showed that: some of the nucleus became small in Con+Aβ42 and the chromatin fracture,while nuclear agglutination significantly increased in RNAi group with an intense fluorescence.Relative to the Con+Aβ42 group,the ratio of nuclear agglutination in Con+DHT+Aβ42group reduced.Relative to the RNAi+Aβ42 group,nuclear gathered slightly decreased in RNAi+DHT+Aβ42,but still more than Con+DHT+Aβ42 group.5.The role of interference CD147 expression in the progress of DHT against the damage about Bax and Bcl-2 protein induced by Aβ42 in SH-SY5Y cellsCD147 Western blot results showed: compared with Con+Aβ42 group(1.21±0.13),the level of CD147 was obviously reduced in RNAi+Aβ42 group(0.40 ± 0.07)and RNAi+DHT+Aβ42(0.40 ± 0.03)(P<0.01),while the Con+DHT+Aβ42 group(1.68 ± 0.05)increased(P<0.05).There was no obvious change between RNAi+Aβ42 group(0.40 ± 0.07)and RNAi+DHT+Aβ42 group(0.40 ± 0.03)(P>0.05).Compared with Con+DHT+Aβ42 group,RNAi+DHT+Aβ42 CD147 was obviously reduced(P<0.01).Bax Western blot results showed that: compared with Con+Aβ42 group(0.41 ± 0.04),the Bax level enhanced in RNAi+Aβ42 group(0.48 ± 0.03)(P<0.05),while the level reduced in Con+DHT+Aβ42group(0.26±0.02)and RNAi+DHT+Aβ42 group(0.34±0.03)(P<0.05).Compared with RNAi+Aβ42group(0.48±0.03),the Bax level reduced in RNAi+DHT+Aβ42 group(0.34±0.03)(P<0.05).Compared with Con+DHT+Aβ42 group(0.26 ± 0.02),RNAi+DHT+Aβ42 group(0.34±0.03)increased significantly(P<0.05).Bcl-2 Western blot experiments results showed that: compared with Con+Aβ42 group(0.91 ± 0.02),the Bcl-2 expression level reduced in RNAi+Aβ42 group(0.63±0.05)and RNAi+DHT+Aβ42 group(0.77±0.05)(P<0.01),while Con+DHT+Aβ42group(1.08 ± 0.12)elevated(P<0.01).Compared with RNAi+Aβ42 group(0.63 ± 0.05),the level of Bcl-2 in RNAi+DHT+Aβ42 group(0.77 ± 0.05)enhanced(P<0.01).Compared with Con+DHT+Aβ42 group(1.08±0.12)the level of Bcl-2 significantly reduced in RNAi+DHT+Aβ42 group(0.77±0.05)(P<0.01).Conclusions:1.DHT can cause the increase of CD147 protein expression in SH-SY5Y cells and flutamide can inhibit the protein increase of CD147 induced by DHT.The promoter region of CD147 exists androgen receptor binding site.2.Interference of CD147 can accelerate damage SH-SY5Y cell vitality induced by Aβ42,hinder the clearance of extracellular Aβ42,accelerate damage SH-SY5Y cell morphology,increase Bax protein level and reduce Bcl-2protein level in the progress of damage induced by Aβ42.3.Interference of CD147 can reduce the cell vitality protection,clear capacity and cell morphology protection of DHT in the progress of against the damage induced by Aβ42.Interference CD147 affects the protein regulation of DHT in the progress of against the damage induced by Aβ42,resulting increasing Bax protein level and reducing Bcl-2 protein level. |
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