ABSTRACT
Objective: To construct the rat Pdcd4 knockdown model of programmed cell death 4 (Pdcd 4) in vivo for further study of the gene's functions. Methods: E3 rats were intranasally instilled with saline or plasmids during antigen induced pulmonary inflammation model induction. The rats were anaesthetized and killed on day 21. The lungs in each group were lavaged by instillation and withdrawal of 2 mL ice-cold PBS for three times through the tracheal route, and the bronchoalveolar lavage fluid (BALF) was collected. After centrifugation, the BALF cell pellet was used to isolate the total RNA. And the mRNA level of Pdcd4 was assessed by real-time quantitative PCR. Results: Pdcd4 expression decreased significantly in BALF cells at the mRNA level after Pdcd4 RNAi in vivo. Conclusion: Rat Pdcd4 knockdown model in vivo was constructed successfully, which can lay foundation for further studies on the functions and mechanism of Pdcd4 gene.
ABSTRACT
Objective To study the significance of HSP47 gene in keloid formation after in vivo study with RNAi technology and the recombinant HSP4 7 siRNA against heat shock protein 47 to keloid in a nude mice model.Methods We injected RNAi mixture into the keloid of a nude mice model in experimental group and PBS water(0.25 ml)into control group at the 16th days after establishing the models.After interference we observed the specimens and harvested specimens at 7th days for biochemical and pathological analysis.Results The expression of HSP47 mRNA reduced obviously and the collagen content also reduced in the experimental group.The rusults had statistical significance.Conclusion We can suppress the expression of HSP47 gene and then reduce the production of collagen after in vivo interfering experiment with HSP4 7 siRNA in keloid nude mice models using RNAi technique.This study cornfirms the mechanism that HSP47 promotes the keloid formation,which provides a new target to treat keloid.