Inhibition effect of mouse orbital fibroblasts TLR4 gene silencing on the thyroid-associated ophthalmopathy

Abstract

AIM:To construct shRNA lentivirus interference vector of mice orbital fibroblasts TLR4 and to research the therapeutic effect and mechanism of TL4-/-fibroblasts on thyroid-associated ophthalmopathy.METHODS:Optimal shRNA interference expression plasmid of mouse orbital fibroblasts TLR4 gene was designed, built, and screened. Then the best shRNA was introduced into lentiviral expression vector by Gateway method and recombinant lentiviral vector was used to infect mouse orbital fibroblasts. Its capability of negative regulating the immune inflammatory response was researched. At last the method of fibroblast TLR4 gene silencing in the model mice of thyroid-associated ophthalmopathy was used, the in vivo therapeutic effect was observed. RESULTS: ShRNA sequences with the best effect of gene silencing were selected and introduced into lentiviral vectors(virus titer was 1.5×106TU/mL). Balb/c mice orbital fibroblasts transfected lentivirus could negatively regulate the immune response, inhibit immune inflammatory response. The proceeding of thyroid-associated ophthalmopathyof the mice transfected TLR4-/-recombinant lentivirus was obviously prior to that of the control mice.CONCLUSION: Mouse fibroblast TLR4-/- siRNA lentiviral vectors are successfully obtained, which has the favourable inhibitory effect on immune inflammatory responses. The recombinant lentivirus could protect the proceeding of thyroid-associated ophthalmopathy, therefore the TLR4 expression interference is a novel potential target for thyroid-associated ophthalmopathy.

Keywords

• shRNA
• TLR4
• fibroblast
• thyroid-associated ophthalmopathy
• negative regulation to immune responses