Effect Of TGF-?1 SiRNA In Caspase-1 And IL-1? Of HSC

Objective : To investigate the expression of caspase-1,IL-1? in HSC-T6 after TGF-?1 was knocked down by TGF-?1 siRNA.Methods:The Lentivirus vectors which contained TGF-?1-siRNA were constructed.These constructed Lentivirus-TGF-?1-siRNA vectors and Lentivirus were transfected into HSC-T6 and selected by puromycin.The transfected cell lines were divided into five groups:control group,control+TGF-?1 group,Lentivirus group,Lentivirus-TGF-?1-siRNA group,Lentivirus-TGF-?1-siRNA+TGF-?1 group,respectively.The qRT-PCR and Western-blot were used to detect the expression of caspase-1,pro-IL-1?,IL-1? and COL-? in each group.Collecting the cell supernatant of each group,ELISA was used to detect the expression of IL-1? and IL-18.HSC-T6 were treated with different concentrations of IL-1? for 24 hr,the proliferation was detected by CCK-8.And COL-I mRNA and protein expression were detected by qRT-PCR and Western-blot.Results: The expression of caspase-1 mRNA and protein was significantly higher in Lentivirus-TGF-?1-siRNA group than that inLentivirus group.After 24 hr treatment with TGF-?1,The expression of caspase-1 was lower inLentivirus-TGF-?1-siRNA+TGF-?1 group than that in Lentivirus-TGF-?1-siRNA group untreated with TGF-?1(P < 0.05).The results of ELISA showed that IL-1 ? in the cell supernatant of Lentivirus-TGF-?1-siRNA group was higher than that in Lentivirus group.After treatment with TGF-?1,the expression of IL-1? was lower in control+TGF-?1 group and Lentivirus-TGF-?1-siRNA+TGF-?1 group than in Lentivirus-TGF-?1-siRNA group untreated with TGF-?1(P < 0.05).There was no significant difference in the expression of IL-18 among different groups.There was no significant difference in expression of IL-1?mRNA in each group.The Western-blot results salso showed that there was no significant difference in the expression of pro-IL-1? among groups.When the HSC-T6 was treated with different concentrations of IL-1?,the proliferation was activated and the expression of COL-?was increased(P < 0.05).And both were positively correlated with IL-1?concentration.Conclusions: This result revealed that TGF-?1 inhibited the release of IL-? by inhibiting caspase-1 to the cleavage of pro-IL-1?,had no effect on the expression of pro-IL-?.