Analysis Of HSP40 Expression In Leucocytes Of Aggressive Periodontitis

It has been demonstrated that aggressive periodontitis (AP) is one of the majorreasons to cause the loss of teeth for healthy people. Several studies showed thataggressive periodontits has strong family linkage and related to genetic disorders.Others showed that the dysfunction of cellular immune system including PMN mayinvolve in the pathogenesis of aggressive periodontitis. Howver, the exact pathogenicmechanism is not clear until today. In order to better understanding the pathogenicbasis of aggressive periodontitis at a molecular level, we examed the gene expressionof normal and aggressive periodontitis subjects by using microarray technology.Objective: To explore differentially expressed genes of Peripheral bloodleucocytes in aggressive periodontitis, and investigate the pathopoiesis mechanism ofdysfunctions in leucocytes which may cause aggressive periodontitis in gene level andthe molecular mechanism of this disease, especially to find some gene which isassociated with Aggressive periodontitis.Methods: 30 ml Peripheral blood sample were taken from three patients(10mlper one) and 10 ml from the health control. The leucocytes were isolated, then we canget total RNA and protein from the leucocytes. Total RNA of leucocytes was drawnby one-step method then the product mRNA was purified, reversely transcribed tocDNA. We use real-time fluorescent quantitative PCR(FQ-PCR) to detect the expresslevel of DNAJCll and PTHLH, then choose HSP40 which is protein production from DNAJC11 for western-blot.Result: We choose DNAJC11 and PTHLH which come from the result of genechip for FQ-PCR. As in Aggressive periodontitis DNAJC11 was up-regulated neal4.5 times in mRNA level than health controls, we choose HSP40 which is proteinproduction from DNAJC11 for Western-blot.HSP40 was up-regulated significantly inprotein level.Conclusion: Gene chip is an effective way to explore differentially expressedgenes of peripheral blood leucocytes in aggressive periodontitis,but there are falsepositive and false negative in the result of gene chip.Real-time fluorescentquantitative PCR is a good way to eliminate false positive. Through FQ-PCR andWestern-blot, we can find DNAJC11 is up-regulated in mRNA and protein level ininperipheral blood leucocytes in aggressive periodontitis. This result can provide thetheory basis for further studies of DNAJC11...