Effect of siRNA interference on nerve growth factor in intervertebral disc inflammation rats

ObjectiveTo investigate the inhibition effect of siRNA interference on NGF induced by inflammatory factor IL-6, and IL-1 so as to provide novel targets for clinical treatment of discogenic low back pain.MethodsThe intervertebral disc nucleus and annulus fibrosus cells of rats were separated. The cells were co-cultured with different concentrations (10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L) of IL-6 and IL-1β. The NGF-siRNA was leaded into the co-cultured cells with its import ability assessed by flow cytometry instrument tests, before and after which the NGF mRNA expression was detected by real-time Q-PCR and the NGF content was detected by ELISA.ResultsFlow cytometry instrument test results showed that the NGF-siRNA cell conversion rate was 99.8%. Real-time Q-PCR detection results showed that compared with negative control group, the NGF mRNA expression of co-cultured cells treated by 10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L IL-6 and IL-1β were respectively raised 3.4, 3.7, 4.7, 3.7 times which were all significantly down-regulated after the import of NGF-siRNA. EILSA detection results showed that compared with negative control group, the NGF content of co-cultured medium treated by 10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L I-L6 and IL-1β were respectively raised 2.9, 3.3, 4.5, 7.4 times which were all significantly decreased after the import of NGF-siRNA.ConclusionsThese molecular biological results suggest that inflammatory factor IL-6 and IL-1β could stimulate NGF on intervertebral disc cells in vitro culture model and its efficiency is concentration dependent, while siRNA interference can inhibit the stimulation effect of IL-6 and IL-1β on intervertebral disc cell, which provides a new targets for the clinical treatment of discogenic low back pain.