Relationship Between Expression Of Zinc Finger Protein A20 MRNA And Apoptosis In Spinal Muscular Atrophy

Spinal muscular atrophy(SMA) is a autosomal recessivetransmission neuromuscular disorder.It's virulence gene was surrivalmotor neuron gene which encoded SMN protein. When the gene wasabsent, full-length surrival motor neuron protein reduced.The histonede-acetylation enzyme inhibitor—valproic acid(VPA) could activategene transcription.So it could activate the express and the transcription ofSMN protein.The degeneration and necrosis of cornu anterius medullae spinalismotoneuron were major patho-change.Apoptosis played an important rolein nerve cells' death. Caspase-3 was at the downstream of apoptotic linearprocess and led to DNA cutting. The SMN gene and zinc finger proteinA20 could inhibit the activity of caspase3 and apoptosis.Objective: We observed the relation between expression ofA20,SMN,Caspase-3mRNA and apoptosis to explore the relationbetween A20 and apoptosis in SMA.Also we would explore thepathogenesy of SMA.The research was to establish experimentfoundation on SMA therapeutics. Method:1 We separated MSCs with density gradient separation and thencultivated them. We appreciated them with CD34 and CD44 byimmunofluorescence staining method.2 We derivated MSCs into neuron-like cells with basic fibroblastgrowth fact, dimethyl sulphoxide and butylated hydroxyanisole. Then weappreciated them with NSE and NF by immunofluorescenee stainingmethod. At the same time,We observed their morphologic change.3 We drew the cell auxodrome to observe the change of cellproliferation.4 Observing the apotosis by TUNEL fluorescein stain.5 Applying RT-PCR to detent the expression of A20, SMN andCaspase-3 mRNA in SMA NLCs and the control NLCs respectively.6 Detecting the fixing information of A20,SMN and Caspase-3.Result:1 The primary cultural MSCs after been disconnected andpurificated displayed that CD34 was negative and CD44 was positive byimmunofluorescence staining method.2 NLCs displayed that majority emerged long ecphyma andcancellated connection.We had accredited them with NSE and NF. Theywere both positive. 3 Every group had two negative-proliferation process.In the secondnegative-proliferation process,SMA NLCs were more manifest comparedwith the control group NLCs (P＜0.05).The negative-proliferationreduced after VPA treatment(P＜0.05).4 The apoptotic rate of SMA NLCs was higher than the controlgroup normal NLCs(P＜0.05).The apoptotic rate of SMA NLCsdecreased(P＜0.05) and that of the control group NLCs increased aftergiven VPA (P＜0.05).5 The expression of A20 and fl-SMN mRNA in SMA NLCs werelower than the control NLCs (P＜0.05). The expression of△7-SMN andCaspase-3mRNA in SMA NLCs were higher than the control NLCs(P＜0.05).The expression of A20,fI-SMNmRNA (P＜0.05) and△7-SMNmRNA(P＞0.05) rose in VPA-treated SMA NLCs.Theexpression of Caspase-3 mRNA reduced in VPA-treated SMA NLCs(P＜0.05). The expression of A20mRNA rose in VPA-treated controlNLCs reduced ((P＞0.05).The expression of fl-SMN,△7-SMN andCaspase-3 mRNA rose in VPA-treated control NLCs.As the VPAconcentration rose,the expression of A20,fl-SMN and△7-SMNmRNArose in VPA-treated SMA NLCs.But the expression of Caspase-3 mRNAdecreased first,and then increased slightly.6 A20 protein had no common fixing relation with fl-SMNprotein.A20 protein had no common fixing relation with Caspase-3 protein by copolymerization microscope.Conclusion:1 In SMA,A20 had anti-apoptotic effect. A20 protein and SMNprotein,A20 protein and Caspase-3 protein maybe have no directinteraction.2 The expression increase of A20mRNA may be one ofanti-apoptotic mechanism of VPA in SMA.