| 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(?-amyloid,A?)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 A? 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 A? plays an important role in the process of AD.Due to the abnormal metabolism,the balance between generation to remove of A? was broken,triggering A? deposition.A variety of enzymes played a significant role in this dynamic process,including ?-,?-secretase related to generating and the NEP,IDE,MMP-2/9 related to degradation,etc.,which maintain the balance in the content of A? 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 5?-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 17?-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 A? 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 A? oviously increased,which showed that the level of brain tissue A? was inversely proportional to the level of testosterone.As previously mentioned,androgen adjust A? level through direct AR pathways or indirect ER pathway.Testosterone can adjust the A? level eather by reduce A? production or by increase the A? remove.There are two main competing cracking way for A? precursor(Amyloid precursor protein,APP): APP transformes into A? thought the action of ?-and ?-enzymes,the other way dose not produce amyloid flexible through ?-secretase,which prevents the generation of A?.Testosterone adjusts the the APP originating from cerebral cortex neurons and reduces the generation of A? by increases non-amyloidosis APP fragment secretion.A? 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 A?,which could be recovered by DHT replacement therapy,suggestting that androgen play a role in regulation of A? by NEP.Androgen may also adjust A? 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 A? and mediation in AD,but its specific molecular mechanism is still not very clear.The research shows that CD147 is another subunit of ?-secretase complexs,participate in the negative regulation ?amyloid precursor protein(APP)to produce A?,and participate in the remove of A?.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 A?42 in SH-SY5Y cellsObjective: Take SH-SY5Y cells as the research object to observ the changes of cell vitality,morphology,apoptosis related proteins and A?42 in the cell culture medium supernatant,following by interfered with LV-BSG-RNAi lentivirus and exogenous A? cell damage.To investigate the relationship among CD147,A?42 clearance and cell damage.Methods:1.The effect of A?42 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 ?M A?42 treatment group(1?M),10 ?M A?42 treatment group(10 ?M),20 ?M treatment group(20 ?M)and 40 ?M A?42 treatment group(40 ?M).The cells were cultured in 100 ?l serum free medium containing different A?42 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 ?l 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 A?42 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 A?42 clear in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+A?42 group(Con+A?42),interference(RNAi),interference+A?42 group(RNAi+A?42).Based on the results of part one,we administrated A?42 of 10 ?M for 24 h and then collected the cell supernatant to test the changes of A?42 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+A?42 group(Con+A?42),interference(RNAi),interference+A?42 group(RNAi+A?42).The cells were cultured in 100 ?l serum free medium containing 10 ?M A?42.After disposure24 h,each hole was added MTS kit reagent 20 ?l 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+A?42 group(Con+A?42),interference(RNAi),interference+A?42 group(RNAi+A?42).The cells were incubated with 10 ?M A?42 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 A?42 in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+A?42 group(Con+A?42),interference(RNAi),interference+A?42 group(RNAi+A?42).The cells were incubated with 10 ?M A?42 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 A?42 in SH-SY5Y cellsExperiment choosed infected cells after 3 d and the cells were randomly divided into control group(Con),control+A?42 group(Con+A?42),interference(RNAi),interference+A?42 group(RNAi+A?42).The cells were incubated with 10 ?M A?42 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 A?42 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 ?M A?42.Compared with the control group(0.80±0.04),1 ?M group(0.75±0.01)did not change obviously(P>0.05).The rest of other groups were 10 ?M 0.65±0.01,20 ?M group 0.59±0.01 and 40?M group 0.48±0.01,respectively(P<0.05).Subsequent experiments were selected 10 ?M A?42 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 A?42 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 A?42 and the A?42 content in culture medium supernatant was lower A?42,with no obvious changes between the thess two groups(P>0.05).the Con+A?42(72.94±4.64)and RNAi+A?42(92.48±6.29)groups were added the exogenous A?42 and the A?42content in culture medium supernatant was higher(P>0.05).,Compared to Con+A?42 group(72.94±4.64),the A?42 content was obviously higher in RNAi+A?42 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+A?42 group(0.57±0.06)and RNAi+A?42 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+A?42 group(0.57±0.06)and RNAi+A?42 group(0.39±0.03)(P<0.01).Compared with Con+A?42 group(0.57±0.06),the cell vitality declined obviously in RNAi+A?42 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 ?M A?42 for 24 h,some of cell showed soma retraction,loss of refraction,emerge particle and axial dinimution in Con+A?42 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 A?42 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+A?42 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+A?42 group.7.The role of interference CD147 expression for the regulation of apoptosis related proteins due to A?42 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+A?42 group(0.21±0.05)were significantly reduced(P<0.01).But there was no obviously change in Con+A?42 group(0.89±0.06)(P>0.05).There was no obviously change between RNAi group(0.23±0.02)and RNAi+A?42 group(0.21±0.05)(P>0.05).Compared with Con+A?42 group(0.89±0.06),the protein level of CD147 in RNAi+A?42 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+A?42 group(1.83 ± 0.19),RNAi+A?42 group(2.32 ± 0.20)(P<0.01).Compared with RNAi group(1.97±0.34),Con+A?42 group(1.83±0.19)had no obvious change(P>0.05),RNAi+A?42group(2.32 ± 0.20)increased significantly(P<0.01).Compared with Con+A?42group(1.83 ± 0.19),the level of Bax increased significantly in RNAi+A?42 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+A?42 group(0.64 ± 0.03),RNAi+A?42 group(0.33 ± 0.02)(P<0.01).Compared with RNAi group(0.66±0.02),Con+A?42 group(0.64±0.03)had no obvious change(P>0.05),RNAi+A?42 group(0.33 ± 0.02)significantly reduced(P<0.01).Compared with Con+A?42 group(0.64±0.03),the level of Bcl-2 reduced in RNAi+A?42 group(0.33±0.02)(P< 0.01).Part three Extracellular matrix metalloproteinase inducer takes part in the regulation of dihydrotestosterone against A?42 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 A?42 in the cell culture medium supernatant,following by interfered with LV-BSG-RNAi lentivirus and exogenous A? cell damage with or withour pretreated DHT.To investigate the role of CD147 in the progress of DHT against the cell damage induced by A?42.Methods:1.The role of interference CD147 expression in the progress of DHT against the damage about cell vitality induced by A?42 in SH-SY5Y cellsExperiment choosed cells with a degree of 60-70% fusion and the cells were randomly divided into control group(Con+A?42),interference group(RNAi+A?42),control group plus RNAi group(Con+DHT+A?42)and interference group plus DHT(RNAi+DHT+A?42).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+A?42 group and RNAi+DHT+A?42 group were replaced by serum free medium containing 100 nm DHT,while the culture medium from Con+A?42 group and RNAi+A?42group 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 ?M A?42 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 A?42 in SH-SY5Y cellsThe experimental group and drug delivery way were the same as experiment 1 A?42 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 A?42 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 A?42 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 A?42 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 A?42 in SH-SY5Y cellsMTS results showed that: compared with Con+A?42 compared group(0.49 ± 0.03),the leve of cell vitality of RNAi+A?42 group(0.34 ± 0.02)significantly decreased(P<0.05),but in Con+DHT+A?42group(0.63±0.03)and RNAi+DHT+A?42 group(0.56 ± 0.03)increased(P<0.05).Compared with RNAi+A?42 group(0.34 ± 0.02),the level of cell vitality of Con+DHT+A?42 group(0.63 ± 0.03)and RNAi+DHT+A?42 group(0.56 ±0.03)increased significantly(P<0.05).Compared with Con+DHT+A?42 group(0.63 ± 0.03),the cell vitality of RNAi+DHT+A?42 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 A?42 in SH-SY5Y cellsElisa results showed that: compared with Con+A?42 group(65.40±4.16),the level of A?42 in RNAi+A?42 group(89.48±3.40)significantly increased(P<0.01),but the data in Con+DHT+A?42 group(35.37 ± 4.32)and RNAi+DHT+A?42 group(53.13 ± 2.34)significantly reduced(P<0.01).Compared to RNAi+A?42 group(89.48 ± 3.40),the level of A?42 in Con+DHT+A?42 group(35.37±4.32)and RNAi+DHT+A?42 group(53.13±2.34)significantly decreased(P<0.01).Compared with Con+DHT+A?42group(35.37±4.32),the level of A?42 in RNAi+DHT+A?42 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 A?42 in SH-SY5Y cellsThe cells from Con+A?42 group showed cell body retraction,loss of refraction,emerged particles and thin axon;most of the cell from RNAi+A?42group 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+A?42 group showed fusiform,triangular and some of them retract.Most cells from RNAi+DHT+A?42 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 A?42 in SH-SY5Y cellsHoechst33258 stainning showed that: some of the nucleus became small in Con+A?42 and the chromatin fracture,while nuclear agglutination significantly increased in RNAi group with an intense fluorescence.Relative to the Con+A?42 group,the ratio of nuclear agglutination in Con+DHT+A?42group reduced.Relative to the RNAi+A?42 group,nuclear gathered slightly decreased in RNAi+DHT+A?42,but still more than Con+DHT+A?42 group.5.The role of interference CD147 expression in the progress of DHT against the damage about Bax and Bcl-2 protein induced by A?42 in SH-SY5Y cellsCD147 Western blot results showed: compared with Con+A?42 group(1.21±0.13),the level of CD147 was obviously reduced in RNAi+A?42 group(0.40 ± 0.07)and RNAi+DHT+A?42(0.40 ± 0.03)(P<0.01),while the Con+DHT+A?42 group(1.68 ± 0.05)increased(P<0.05).There was no obvious change between RNAi+A?42 group(0.40 ± 0.07)and RNAi+DHT+A?42 group(0.40 ± 0.03)(P>0.05).Compared with Con+DHT+A?42 group,RNAi+DHT+A?42 CD147 was obviously reduced(P<0.01).Bax Western blot results showed that: compared with Con+A?42 group(0.41 ± 0.04),the Bax level enhanced in RNAi+A?42 group(0.48 ± 0.03)(P<0.05),while the level reduced in Con+DHT+A?42group(0.26±0.02)and RNAi+DHT+A?42 group(0.34±0.03)(P<0.05).Compared with RNAi+A?42group(0.48±0.03),the Bax level reduced in RNAi+DHT+A?42 group(0.34±0.03)(P<0.05).Compared with Con+DHT+A?42 group(0.26 ± 0.02),RNAi+DHT+A?42 group(0.34±0.03)increased significantly(P<0.05).Bcl-2 Western blot experiments results showed that: compared with Con+A?42 group(0.91 ± 0.02),the Bcl-2 expression level reduced in RNAi+A?42 group(0.63±0.05)and RNAi+DHT+A?42 group(0.77±0.05)(P<0.01),while Con+DHT+A?42group(1.08 ± 0.12)elevated(P<0.01).Compared with RNAi+A?42 group(0.63 ± 0.05),the level of Bcl-2 in RNAi+DHT+A?42 group(0.77 ± 0.05)enhanced(P<0.01).Compared with Con+DHT+A?42 group(1.08±0.12)the level of Bcl-2 significantly reduced in RNAi+DHT+A?42 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 A?42,hinder the clearance of extracellular A?42,accelerate damage SH-SY5Y cell morphology,increase Bax protein level and reduce Bcl-2protein level in the progress of damage induced by A?42.3.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 A?42.Interference CD147 affects the protein regulation of DHT in the progress of against the damage induced by A?42,resulting increasing Bax protein level and reducing Bcl-2 protein level. |
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