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Objective To investigate the effect of tripterygium glycosides (TG) contained serurn on the pathological boneforming related inflammatory markers and miR-21.Methods Previous isolated and cultured AS fibroblasts were stimulated using IL-1 of 1ng/ml for 24h,different concentrations of blank serum (5%,10%,and 15%) and TG contained serum (5%,10%,and 15%) were added for 48h.PGE-2,IL-17,IL-22,IL-23,CCL19 and CCL21 proteins were examined by Western blot.The osteogenesis marker BMP and microRNA-21 mRNAs were tested.Results 48 h after intervention,the expressions of inflammatory markers were obviously inhibited by TG contained serum;the boncforming related inflammatory markers,expressions of BMP-2 and miR-2t were all inhibited in a dose-dependent manner.Conclusions TG could inhibit the expressions of boneforming related inflammatory markers,BMP-2 and miR-21,thus providing theoretical basis to treat AS pathological boneforming.
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Transplantation of mesenchymal stem cells [MSCs] can promote functional recovery of the brain after hypoxic-ischemic brain damage [HIBD]. However, the mechanism regulating MSC migration to a hypoxic-ischemic lesion is poorly understood. Interaction between stromal cell-derived factor-1 alpha [SDF-1 alpha] and its cognate receptor CXC chemokine receptor 4 [CXCR4] is crucial for homing and migration of multiple stem cell types. In this study, we investigate the potential role of SDF-1 alpha /CXCR4 axis in mediating MSC migration in an HIBD model. In this experimental study, we first established the animal model of HIBD using the neonatal rat. Bone marrow MSCs were cultured and labeled with 5-bromo-21-deoxyuridine [BrdU] after which 6×10[6] cells were intravenously injected into the rat. BrdU positive MSCs in the hippocampus were detected by immunohistochemical analyses. The expression of hypoxia-inducible factor-1 alpha [HIF-1 alpha] and SDF-1 alpha in the hippocampus of hypoxic-ischemic rats was detected by Western blotting. To investigate the role of hypoxia and SDF-1 alpha on migration of MSCs in vitro, MSCs isolated from normal rats were cultured in a hypoxic environment [PO[2]=1%]. Migration of MSCs was detected by the transwell assay. The expression of CXCR4 was tested using Western blotting and flow cytometry. BrdU-labeled MSCs were found in the rat brain, which suggested that transplanted MSCs migrated to the site of the hypoxic-ischemic brain tissue. HIF-1 alpha and SDF- 1 alpha significantly increased in the hippocampal formations of HIBD rats in a time-dependent manner. They peaked on day 7 and were stably expressed until day 21. Migration of MSCs in vitro was promoted by SDF-1 alpha under hypoxia and inhibited by the CXCR4 inhibitor AMD3100. The expression of CXCR4 on MSCs was elevated by hypoxia stimulation as well as microdosage treatment of SDF-1 alpha . This observation illustrates that SDF-1 alpha /CXCR4 axis mediate the migration of MSCs to a hypoxic-ischemic brain lesion in a rat model
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Animals, Laboratory , Receptors, CXCR4 , Mesenchymal Stem Cells , Hypoxia-Ischemia, Brain , Rats , Models, AnimalABSTRACT
Objective To explore the role of HIF-1 and its downstream SDF-1α/CXCR4 and VEGF/VEGFR pathway in mediating MSC mobilization with DMOG .Methods Male SD rats were randomly divided into five groups:Normal saline control group , DMOG group, YC-1 group, AMD3100 group, SU5416 group.We used CFU-F assay and flow cytometry to determine the number of MSCs in rat bone marrow ( BM ) and peripheral blood ( PB ) in each group , respectively.The concentrations of SDF-1αand VEGF both in BM and PB serum in each group were detected by ELISA . Western blotting was used to test protein levels of HIF-1α, SDF-1αand VEGF in BM.Results Compared with NS group, the number of CFU-Fs as well as the percentage of CD 45 -CD90 +cells increased in DMOG group ( P <0.05);Compared with DMOG group, the number of CFU-Fs as well as the percentage of CD 45 -CD90 +cells decreased in YC-1 group, AMD3100 group and SU5416 group (P <0.05).Compared with DMOG group, the concentration and protein expression of HIF-1αdecreased significantly in YC-1 group ( P <0.05 ) , the concentration and protein expression of SDF-1αdecreased significantly in AMD 3100 group ( P <0.05 ) , the concentration and protein expression of VEGF decreased significantly in SU5416 group ( P <0.05 ).Conclusion DMOG can induce MSCs mobilization possibly via up-regulating the expression of HIF-1αand activating its downstream SDF-1α/CXCR4 and VEGF/VEGFR pathway .
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Objective To explore the expression of Ang -2 and CD105 in breast cancer and their po-tential associations with the cancerization and progression of breast cancer .Methods Thirty patients with breast cancer ( cancerous tissue ) ,15 intraductal papilloma cases ( precancer tissue ) and 15 normal breast tissue were col-lected for each group ,respectively .Immunohistochemistry ,western blot and RT-PCR techniques were used to ex-amine the expressions of Ang -2 and CD105 in cancerous breast tissue ,precancer breast tissue and normal breast tissue at both protein and mRNA levels .The possible correlations of Ang -2 and CD105 expression with clinico-pathological characteristics of the breast cancer were analyzed .Results The expressions of CD105 and Ang-2 in cancer and precancer tissues were higher than in normal tissues at both protein and mRNA levels ,and the trend of their expressions was increased from normal to precancer ,and cancer tissue .The expressions of CD 105 and Ang-2 protein and mRNA were positively correlated with tumor size ,invasion,lymph node metastasis,and negatively correlated with the differentiation of the cancer .Conclusion CD105 and Ang-2 are involved in the canceriza-tion and pregression of breast cancer .These two biomarkers can serve as two useful indicators in assisting diagno-sis and prognosis of breast cancer .
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Objective To explore the role of MMP-9 in blood-brain barrier (BBB) disruption and to promote rMSCs migration.Methods To investigate the effect of MMP-9 on the permeability of BBB,an in vitro model of BBB was established and performed Transwell experiments.To observe the pathologic changes of the cerebral cortex,rat brains from Sham treated group,HIBD group,MMP-9 treated group and TIMP-1 (intervention) treated group at different time points (0.5,1,3,7,14 d) were prepared and stained with Hematoxylin-Eosin and quantified the brain water content.Permeability of BBB examined by EB values measurement.MMP-9 protein level of rat cortex was detected by Western Blot.The number of Brdu labeled rMSCs in the rat cerebral cortex of each group was quantified by Immunohistochemistry (IHC).Results Transwell experiments results showed that migration of rMSCs increased remarkably in hypoxic condition compared to that of normal control (P<0.01).The number of rMSCs migrated in the MMP-9 treated group was much more than that of negative control group and TIMP-1 group (P<0.01).The results of pathology showed that compared to HIBD group,brain water content and the permeability of the BBB were increased in MMP-9 treated group but reduced in TIMP-1 treated one.The expression of MMP-9 protein in MMP-9 treated group was reached the peak at 3 d point,which was higher than that of HIBD group and TIMP-1 group (P<0.05,P<0.01).The quantity of Brdu labeled rMSCs crossed BBB in MMP-9 treated group was extremely higher than that of HIBD group (P<0.05,P<0.01).Conclusions The expression level of MMp-9 protein was improved after hypoxic-ischemic.MMP-9 could facilitate the migration of rMSCs through vitro BBB and control the opening of BBB,which indicate that MMP-9 may facilitate the migration of rMSCs through BBB into brain.