Effects Of Jianpi Bushi Recipe On Matrix Metalloproteinase Activation And NF-κB Pathway In Olfactory Ensheathing Cells Of Hypoxia/Reoxygenation Injury Model

Objective:To investigate the effect of Strengthening Spleen prescription on influence and mechanism of activation of matrix metalloproteinases and NF-κB signal path in olfactory ensheathing cells with hypoxia/reoxygenation injury.Methods:Newborn<1d of SD rats were removed under sterile conditions with trypsin after bilateral olfactory bulb olfactory ensheathing cells after digestion to obtain a single cell suspension by differential adherent cytarabine nerve growth factor methods of olfactory ensheathing cells were purified and identified; explore non-toxic concentrations of serum containing a combination of MTT, the maximum non-toxic concentrations of the drug-containing serum concentrations of experimental work as cytology concentration(15%); olfactory ensheathing cells were randomly divided into five groups:A:normal serum control group, B:normal serum+hypoxia (10% O2) group, C:hypoxia edaravone containing serum group, D:Strengthening Spleen prescription contained serum group, E:normal serum+hypoxia+PDTC (NF-κB inhibitor,50μmol/L) group, the plates were placed into carbon dioxide incubator within an anaerobic box, bag added micro-aerobic capacity and oxygen indicator, after induction 24,48, return to normal culture conditions 4h, resulting in hypoxia/ reoxygenation. When compared to the end of each experimental group were MMP-2, MMP-9, TIMP-1, TIMP-2 protein expression; cytoplasm nucleus P65, and P50 of expression; cytosolic IκBα, P-IκBα, IκKβ of expression; NF-κB DNA binding activity.Results:(1) After experimental hypoxia group at each time point of MMP-2, MMP-9 protein/the cytoplasm and nucleus of P65, P50/the cytoplasm P-IκBα, IκKβ/NF-κB DNA binding activity expression were significantly increased, the difference was statistically significant (P<0.01); compared with hypoxia group, western medicine, traditional Chinese medicine group, PDTC group at each time point of MMP-2, MMP-9 protein expression were significantly lower, the difference was statistically significant (P<0.05).(2) After experimental hypoxia group at each time point TIMP-1, TIMP-2 protein and cytoplasmic IκBα expression were significantly lower, the difference was statistically significant (P<0.01); compared with hypoxia group, Western medicine group, TCM group, PDTC group at each time point TEMP-1, TIMP-2 protein and cytoplasmic IκBα expression were significantly increased, the difference was statistically significant (P<0.05).Conclusions:(1) Strengthening Spleen prescription in order olfactory ensheathing cells by inhibit the expression of MMP-2, MMP-9, and the promotion of TIMP-1, TIMP-2’s expression after reoxygenation injury;(2) Strengthening Spleen prescription inhibit excessive expression of olfactory ensheathing cells from hypoxia/reoxygenation injury P65, P50, P-IκBβ, IκKβ, and promote the expression of IκBα, thus reducing NF-κB DNA binding activity, inhibition of NF-κB activation.(3) Strengthening Spleen prescription in inhibiting olfactory ensheathing cells MMP-2 expression of hypoxia/reoxygenation injury, MMP-9, and the promotion of TIMP-1, TIMP-2 expression may be related to inhibition of NF-κB activation.