Expression Of Bcl-2 Gene In Peripheral Blood Mononuclear Cells Of Patients With Relapsing Neuromyelitis Optica

Objective1.To approach the expression of Bc1-2 gene in peripheral blood mononuclear cells(PBMC) of patients with Neuromyelitis optica(NMO) and the correlation between Bc1-2 gene expression and relapse of the disease;2.To approach the correlation between the expression of Bc1-2 gene and the relapsing time.Methods1.To select 25 cases of definite NMO patients,and divided into 2 groups:14 cases in relapsed group and 11 cases at rest in remitting group.2.The 5 ml blood samples were used to separate PBMC by density-gradient centrifugation method for all patients.3.It was used by TRIzol method to extract mRNA in PBMC.4.Extracted mRNA was template,and mRNA was to be reverse transcription into cDNA by two-steps reverse transcription-PCR.Bc1-2 as target gene and GAPDH as inter control,two genes were respectively amplified through guidance of their primers.5.Amplified products were stained by Ethidium bromide,and observed by 1% agarose gel electrophoresis,also taken photos by gel imaging analyzer.6.Statistical data was the gray scale ratio of Bc1-2 gene and GAPDH gene in same sample.QUANTITY ONE was mainly used to process test pictures,and GENE TOOLS to confirm them.Data come from two software were statistical analysis by appropriate statistical methods in SPSS.Results1.Patients in relapsed group compared general information with remitting group,the age differences(t = 0.216,P＞0.05),gender differences(x~2=0.365,P＞0.05), differences in numbers of relapsing time(t=0.275,P＞0.05) were not statistically significant. 2.The expression of Bc1-2 gene in peripheral blood mononuclear cells of patients in relapsed group is statistically significantly higher than that in remitting group (t=6.096,P＜0.001).3.Based on the relapsing time,relapsed group can divided into＜2 days,2-4 days,4-6 days and≥6 days subgroups.Among the four subgroups,differences in Bc1-2 gene expression in peripheral blood mononuclear cells were statistical significant(F = 70.880,P＜0.001).4.Comparison between Subgroups showed as follow:Bc1-2 gene expression of the subgroup in 2 days after relapse was statistically significantly higher than that in other subgroups,including the 2-4 days subgroup,4-6 days subgroup and≥6 days subgroup(all P＜0.001).2-4 days subgroup,compared with 4-6 days subgroup,there was no statistical significance in difference(t=1.819,P＞0.05);and compared with≥6 days subgroup,there was a significant statistical significance(t=5.085,P＜0.01). Also,the contrast was not statistical significant between 4-6 days subgroup and≥6 days subgroup(t=2.766,P＞0.05).5.It can get test dada with processing test pictures by QUANTITY ONE and GENE TOOLS independently.Two groups of dada which come from different softwares came to the same statistical results after analysis by SPSS.All the data above was deal with QUANTITY ONE as the example.Conclusions1.There was detected enhancement of Bc1-2 gene expression after NMO relapse.So we could come to the conclusion that the relapse of NMO is relational with the expression of Bc1-2 gene in peripheral blood mononuclear cells.2.The enhanced extent of Bc1-2 gene in peripheral blood mononuclear cells was relational with the relapsing time.This tend was more significantly in the initial stage of relapse,and to weaken gradually with prolong of time after relapse.