ABSTRACT
Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.
ABSTRACT
Objective To evaluate the effects of astragalus on liver injury and TGF-β1 mRNA expression in immature rats with obstructive jaundice(OJ).Methods Forty immature male Wistar rats were randomly divided into a control,a shamoperation,an OJ,and an OJ+A groups.Wistar rats in the OJ and OJ+A group were subjectedto common bile duct ligation(CBDL),while the sham group had the bile duct mobilized but not tied.The control,sham,and OJ groups were giyen 0.5ml of normal saline by intrapedtoneal injection daily.In the OJ+A group,250mg/100g body weight of astragalus injection was applied intraperitoneally daily from day 1 to 7 of the study.All animals were sacrificed on postoperative day 8.Liver tissue and peripheral blood were colleered.Serum Alanine aminotransferase(ALT),Aspartate aminotransferase(AST),and total cholic acid(TCA)level were detected.Hepatic morphological findings were observed by light microscopy.TGF-β1 mRNA were extracted from liver and measured by reverse transcription polymerase chain reaction(RT-PCR).Data were analyzed using chi-square test and student's test(-x±s),P＜0.05 was considered statistical significance.Results ①Serum AST,ALT,and TCA value in each OJ group were significantly higher than that of the control and sham operation groups.②In the control and sham operation group,normal structure of the liver remained,but in the OJ group,obvious degeneration of hepatocytes was detected,such as cholastasis,starch accumulation and fatty degeneration.Kupffer cells and dilated Disse'space were common in the OJ group.In the OJ group.histopathologic findings of the liver demonstrated intense degree of fibrosis,but in the OJ+A group.typical changes of large duct obstruction were significantly improved after astragalus injection treatment.③The TGF-β1 mRNA over-expression in the OJ group was seen compared with the control and the OJ+A groups.Conclusion ①In growing rat model of experimental obstructive jaundice,hepatic morphology and liver biochemical tests altered significantly.②The administration of astragaius can ameliorate liver damage and diminish expression of TGF-β1 mRNA of hepatic tissue,thus administration of astragalus may be effective in preventing hepatic injury in growing rats with obstructive jaundice.
