ABSTRACT
Objective · To investigate the expression of the regulator of calcineurin 1 (RCAN1) and calcineurin A (CnA) in tissues of in-stent restenosis after intervention of arteriosclerosis obliterans (ASO), and to explore the relationship between their expression levels and the occurance of in-stent restenosis. Methods · Superficial femoral arterial tissues were collected from 15 ASO patients undergoing lower extremity amputation for in-stent restenosis in Department of Vascular Surgery, The First Affiliated Hospital of Chongqing Medical University from September 2013 to June 2016. H-E staining and Masson staining were performed on the stenosis tissues, as well as on the proximal and distal tissues, and the morphological changes of these tissues were observed under optical microscope. Western blotting was used to detect the protein levels of RCAN1, CnA and proliferating cell nuclear antigen (PCNA). The distribution of RCAN1 and CnA proteins was observed by immunohistochemistry and immunofluorescence methods. In addition, co-immunoprecipitation was used to validate the protein-protein interaction between RCAN1 and CnA in vascular tissues. Results · The expression of RCAN1 in the distal tissues was significantly elevated compared with the proximal tissues and the stenosis tissues (P<0.05). The expression of RCAN1 in the proximal tissues was higher than that in the stenosis tissues (P <0.05). The expression of CnA and PCNA in the stenosis tissues was significantly elevated compared with the proximal tissues and the distal tissues (P<0.05). Immunohistochemistry and immunofluorescence analyses showed that RCAN1 and CN proteins were mainly expressed in the cytoplasm of vascular smooth muscle cells. Co-immunoprecipitation analysis showed there is protein-protein interaction between RCAN1 and CnA in arterial tissues. Conclusion · The low expression of RCAN1 and the high expression of CnA are probably related to the occurrence of in-stent restenosis.
ABSTRACT
[Objective]To forecast the sponge mechanism mediated by LOC389023 in patients with intractable temporal lobe epilepsy(TLE),through investigating the expression of microRNA interacted with dipeptidyl peptidase 10(DPP10)and LOC389023.[Methods]The expression of DPP10 and Kv4.3 were detected in 15 temporal neocortex from patients with brain trauma (control group)and in 26 temporal neocortex from patients with intractable TLE(epilepsy group)by western blot(WB)and immunohisto?chemical(IHC)staining. The location of DPP10 and voltage dependent potassium channel 4.3(Kv4.3)was detected by immunofluo?rescent(IF)staining. The interaction between DPP10 and Kv4.3 was testified by co-immunoprecipitation(Co-IP). The expression of microRNA obtained by softwares(miRanda,Pita,TargetScan and miRDB)was detected by qPCR.[Results]IHC and WB showed an increased expression of DPP10(P0.05). But decreased expression of LOC389023 and miR-140-5p and increased expression of miR-25-3p and miR-367-3p were found in epilepsygroup compared to control group (P < 0.05).[Conclusion]miR-25-3p and miR-367-3p may be regulated by LOC389023 through the sponge mechanism followed by altered expression of DPP10 in intractable temporal lobe epilepsy.