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In order to find out the potential modulators which influence the secretion of TNF α , the relationship between the amount of secreted TNF α (sTNF α ) and the level of TNF α converting enzyme (TACE) gene expression was studied before and after the stimulation by lipopolysaccharides (LPS) to HL 60 cells and adhered cells isolated from human spleen using cytological and molecular biology techniques and methods (RT PCR, Dot blot hybridization, etc.). The experimental results showed that: (1) LPS could induce the increase of expression of TNF α mRNA and TACE mRNA, reaching the peak value at 6 h and 10 h respectively after addition of LPS into cell culture medium; (2) The anti sense oligodeoxyribonucleotide (A ODN) complementary to TACE mRNA sequence really inhibited the secretion of TNF α as a result of blocked translation of TACE gene; (3) Furthermore, it was also observed that RDQ, a kind of injection derived from Chinese traditional herb, had strongly inhibitory effects on the expression of TACE mRNA and secretion of sTNF α stimulated by LPS. The above results suggested that the TACE indeed involved in delivering and processing of pro TNF α during the period of LPS stimulation and the study about the regulator/inhibitors of TACE gene expression would be very important to develop new types of therapy agents against toxic and side effects of sTNFα during the peroid of infection/inflammation to human body.
In order to find out the potential modulators which which the secretion of TNF α, the relationship between the amount of secreted TNF α (sTNF α) and the level of TNF α converting enzyme (TACE) gene expression was studied before and after the stimulation by lipopolysaccharides (LPS) to HL 60 cells and adhered cells isolated from human spleen using cytological and molecular biology techniques and methods (RT PCR, Dot blot hybridization, etc.). The experimental results showed that: (1) LPS could induce the increase of expression of TNFα mRNA and TACE mRNA, the peak value at 6 h and 10 h respectively after addition of LPS into cell culture medium; (2) The anti sense oligodeoxyribonucleotide (A ODN) complementary to TACE mRNA sequence really inhibited the secretion of TNFα as a result of blocked translation of TACE gene; (3) Furthermore, it was also observed that RDQ, a kind of injection derived from Chinese traditional herb, had stron gly inhibitory effects on the expression of TACE mRNA and secretion of sTNF α stimulated by LPS. The above results suggested that the TACE indeed involved in delivering and processing of pro TNF α during the period of LPS stimulation and the study about the regulator / inhibitors of TACE gene expression would be very important to develop new types of therapy agents against toxic and side effects of sTNFα during the peroid of infection / inflammation to human body.