Regulation Of Puerarin On Rat Cervical Intervertebral Disc Cell Apoptosis Induced By IL-1β

Objective:In recent years, the incidence of cervical disease is rising, and the age of onset of a younger age trend, impact on the patient’s physiological and psychological cause many serious impact on patients with normal work and life. The experiment by studying puerarin on apoptosis in rat carotid inflammatory cytokines IL-1β-induced apoptosis of intervertebral disc cells, Bcl-2and Caspase-3of the regulation, to explore its delay disc degeneration, prevention and treatment of cervical spondylosis the mechanism of action, and experimental basis and theoretical basis for the clinical use of Radix treatment of cervical spondylosis.Method:1.Isolated and cultured rat cervical intervertebral disc cells in vitro, stable passaged.2.To identified cells by toluidine blue staining and immunocytochemistry staining.3.Different concentrations of IL-1β-induced rat cervical intervertebral disc cell apoptosis and wst-1detector cell activity, select the appropriate concentration of the intervention..4.Wst-1method to detect different concentrations of puerarin treatment by IL-1β-induced apoptosis in rat cervical intervertebral disc cells vitality.5.To detected the expression levels of Bcl-2and Caspase-3of cells treated by IL-1and different concentration puerarin by Western Blot.Results:1.The cultured cells were stained with toluidine blue metachromasia, Purple stained cytoplasm and nuclei stained blue; Ⅰ、Ⅱ collagen immunocytochemistry results are visible expression:Mostly spindle-shaped cells, no nuclear staining, while the cytoplasm showed brown. The primary cultured cells was identified as rat intervertebral disc cells.2.Different concentrations of IL-1β group and Control group with a significant difference (P<0.05). Different times of the day, the difference between the same intervention group was significantly (P<0.05). With the increase in the dose and the time of intervention of the IL-1β, the apoptotic effect becomes more obvious, when the intervention time to a certain extent, the increase in the concentration of IL-1β and no obvious showing the role to enhance the apoptotic effect.3.The apparent protective effect of high doses of puerarin cells, with IL-1β-induced apoptosis group, a significant difference (P<0.05)4. Caspase-3protein expression levels with puerarin dose increased expression weakened; bcl-2protein expression levels in the normal group and pure IL-1β intervention group than Puererin groups express lower, with a significant difference (P<0.05).Conclusion:1.IL-1β can successfully induce apoptosis of intervertebral disc cells in rat cervical was positively correlated with the dose2. Puerarin by raised anti-apoptotic factor Bcl-2down the apoptotic factor Caspase3protein expression level to block apoptosis signal transduction, which protect cell activity, inhibit excessive apoptosis delay disc degeneration.3.PPuerarin effectively protect annulus fibrosus cells to resist the destruction of inflammation of the fibrous ring to protect the intervertebral disc, delaying the role of disc degeneration.