ABSTRACT
Objective:To study the change of related molecular about apoptosis we reduce the expression of CD59 on acute T lymphocyte Jurkat cell lines .Methods: RNA interference (RNAi) was used to reduce the expression of CD59 gene by lentivirus,confocal was applyed to observe the transfection efficiency and the location of CD59 molecular then Real-time-PCR and Western blot were used to select the most effective group to do the rest experiment;Western blot was used to detect the change of expression about Bcl-2,Bax,Caspase-3 and Survivin;ELISA was used to investigate the expression of IL-3 and TNF-β.Results: Confocal observed each group′s transfection efficiency over 90%,CD59 molecules were mainly located in cell membrane;Real-time-PCR and Western blot showed group A had the best down-regulation efficiency;we defined RNAi-CD59-A as experimental group for subsequent experiments named RNAi-CD59;the experimental group can enhance the expression of Bax,caspase-3 (P<0.05),inhibit the expression of Bcl-2 and Survivin(P<0.05);ELISA showed that the expression of IL-3 in the down-regulation group increase(P<0.05),the expression of TNF-β decrease (P<0.05).Conclusion: Down-regulation CD59 can promote the expression of apoptosis molecular in acute leukemia Jurkat cell lines restrain the expression of proliferation molecular.
ABSTRACT
BACKGROUND:The mechanism underlying Wal erian degeneration fol owing peripheral nerve injury is complex. Immune regulation on Wal erian degeneration is beneficial for early repair of perpheral nerve injury. OBJECTIVE:To investigate the effects of Tol-like receptor 4 (TLR4) antagonist on Wal erian degeneration and axonal regeneration after early peripheral nerve injury in rats. METHODS:Fifty male Wistar rats were recruited and randomly divided into treatment group (n=20), model group (n=20) and sham group (n=10). The right sciatic nerves of rats in treatment and model groups were cut and sutured end-to-end, while the sciatic nerves of rats in sham group were only exposed. In the treatment group rats were intravenously injected with 0.15 mg/kg TAK-242 via tail vein 1 hour preoperatively and 7 days postoperatively, and the rats in the other two groups were given intravenous injection of the same volume of normal saline. The sciatic nerves were removed at 24 hours, 3, 4 and 7 days after surgery. RESULTS AND CONCLUSION:Real-time PCR indicated that the mRNA expressions of interleukin-1βand monocyte chemoattractant-1 were significantly increased in the model group compared with the sham group at 24 hours after surgery (both P<0.001), while the expressions were significantly decreased after TAK-242 injection (both P<0.001). Immunofluorescence showed that compared with the model group, down-regulated expression of CD68+and iba1+cel s appeared in the treatment group at 3 days after surgery (P<0.01, P<0.05). Luxol fast blue staining revealed that demyelination at the sciatic nerve stump appeared in both model and treatment groups at postoperative 7 days, but myelin debris clearance in the treatment group was significantly reduced compared with the model group (P<0.05). Hematoxylin-eosin staining showed that a lot of inflammatory cel s, Schwann cells and regenerated nerve fibers at the sciatic nerve stump were found in the model group, while there were few inflammatory cells, Schwann cel s and regenerated nerve fibers in the treatment group at 7 days after surgery. Immunohistochemistry found that the expression of growth-associated protein-43 in the treatment group was significantly lower than that in the model group at 4 days postoperatively (P<0.05). Besides, compared with the model group, a significantly decreased sciatic functional index was found in the treatment group at 20, 30 and 40 days after surgery (P<0.05). These results show that TLR4 antagonists delay early nerve regeneration in rats after sciatic nerve injury probably by inhibiting the TLR4 signaling pathway.