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Background and Objectives@#Mesenchymal stem cells (MSCs) have the multipotent capacity to differentiate into multiple tissue lineages as well as to self-renew, which is the main origin of adipocytes. IL6/IL6R pathway exerts a significant role in tissue regeneration and cell differentiation. Whereas, the underlying mechanism between IL6/IL6R pathway and MSCs adipogenesis differentiation remains elusive. @*Methods@#MSCs from healthy donors were cultured in adipogenesis differentiation medium for 0∼14 days, during which their adipogenesis differentiation degree was evaluated by Oil Red O staining. The expression of IL6R was detected in MSCs during adipogenesis differentiation. Knockdown and overexpression of IL6R were respectively performed using siRNA and lentivirus to investigate its effect on MSCs adipogenesis differentiation. The adipogenesis marker genes expression and MAPK pathway activation were detected by Western blotting. The role of P38 pathway in the adipogenesis differentiation of MSCs was determined using the specific inhibitor SB203580. @*Results@#The expression of IL6 and IL6R increased during adipogenesis differentiation in MSCs, which were positively correlated with Oil Red O quantification result. Knockdown and overexpression experiments demonstrated a positive correlation between the expressions of IL6R and MSCs adipogenesis differentiation, accompanied by same trend of P38 phosphorylation. Besides, the specific P38 inhibitor SB203580 markedly inhibited the adipogenesis differentiation potential of MSCs. @*Conclusions@#This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.
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Ankylosing spondylitis (AS) is a type of autoimmune disease that predominantly affects the spine and sacroiliac joints. However, the pathogenesis of AS remains unclear. Some evidence indicates that infection with bacteria, especially Gram-negative bacteria, may have an important role in the onset and progression of AS. Recently, many studies have demonstrated that mesenchymal stem cells (MSCs) dysfunction may contribute to the pathogenesis of many rheumatic diseases. We previously demonstrated that MSCs from AS patients exhibited markedly enhanced osteogenic differentiation capacity in vitro under non-inflammatory conditions. However, the properties of MSCs from AS patients in an inflammatory environment have never been explored. Lipopolysaccharide (LPS), a proinflammatory substance derived from the outer membrane of Gram-negative bacteria, can alter the status and function of MSCs. However, whether MSCs from AS patients exhibit abnormal responses to LPS stimulation has not been reported. Autophagy is a lysosome-mediated catabolic process that participates in many physiological and pathological processes. The link between autophagy and AS remains largely unknown. The level of autophagy in ASMSCs after LPS stimulation remains to be addressed. In this study, we demonstrated that although the basal level of autophagy did not differ between MSCs from healthy donors (HDMSCs) and ASMSCs, LPS-induced autophagy was weaker in ASMSCs than in HDMSCs. Specifically, increased TRAF4 expression in ASMSCs impaired LPS-induced autophagy, potentially by inhibiting the phosphorylation of Beclin-1. These data may provide further insight into ASMSC dysfunction and the precise mechanism underlying the pathogenesis of AS.
Subject(s)
Humans , Autoimmune Diseases , Autophagy , Bacteria , Gram-Negative Bacteria , In Vitro Techniques , Membranes , Mesenchymal Stem Cells , Pathologic Processes , Phosphorylation , Rheumatic Diseases , Sacroiliac Joint , Spine , Spondylitis, Ankylosing , Tissue Donors , TNF Receptor-Associated Factor 4ABSTRACT
Objective To investigate the relation between microembolic signals (MES) and vertebral basilar artery ste?nosis in patients with brainstem infarction. Methods A total of 156 patients with acute brainstem infarction, who were de?termined the cerebral infarction lesion and vertebral basilar artery stenosis by cranial magnetic resonance imaging and CT an?giography, and were monitored by transcranial Doppler via occipital window of basilar arterial MES monitoring in 7 days of the onset, were divided into microembolus signal negative group (n=136) and positive group (n=20). The clinical data were compared between two groups. The differences of different degrees of stenosis were analyzed in two groups. The differences of different locations of stenosis in patients with vertebral basilar artery stenosis were analyzed in two groups. Logistic regres?sion analysis was used to analyse the factors affecting MES. Results There were no significant differences in age, gender, history of hypertension and diabetes mellitus between the two groups (P<0.05). There were significant differences in the dif?ferent degrees of stenosis between two groups, no or mild stenosis was found in MES-negative group and severe stenosis in MES-positive group (P<0.05). There were 70 cases with no vertebral basilar artery stenosis, 86 cases with mild, moderate and severe stenosis, in which 14 cases were MES-positive and 72 cases were negative. There were significant differences in different locations of stenosis between the two groups. The proportion of multiple infarctions was significantly higher in MES-positive group than that of MES-negative group (P<0.05). The intracranial vertebral basilar artery stenosis and 75%of ver?tebral basilar artery stenosis were the independent risk factors of MES-positive. Conclusion Severe stenosis of the verte?bral basilar artery is more vulnerable to occur MES of posterior circulation, leading to cerebral infarction. Microemboli may be the cause of multiple infarctions in patients with vertebral basilar artery stenosis.
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BACKGROUND:Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is stil unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE: To explore the immunoregulation capability of bone marrow mesenchymal stem cels from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cels from healthy donors on ankylosing spondylitis. METHODS: Bone marrow mesenchymal stem cels were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cels were used in subsequent experiments. A human monocytic cel line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cels and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6 (TSG-6) proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the mRNA level of cytokines secreted by bone marrow mesenchymal stem cels and macrophages. RESULTS AND CONCLUSION:The typical mesenchymal stem cel surface markers were expressed in both bone marrow mesenchymal stem cels from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and mRNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cels from patients with ankylosing spondylitis were obviously higher than those from healthy donors (P < 0.05). TSG-6 secreted by bone marrow mesenchymal stem cels from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cels from healthy donors in both RNA transcriptional and protein levels (P < 0.05). Our study demonstrates that bone marrow mesenchymal stem cels from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting the tumor necrosis factor-α secretion from macrophages, which reveals a mechanism of immune disorder in ankylosing spondylitis. The therapeutic mechanism of bone marrow mesenchymal stem cels from healthy donors may work by secreting enough TSG-6 to inhibit the activation of macrophages in patients with ankylosing spondylitis, and thereby to decrease the secretion of tumor necrosis factor-α. Cite this article:Sun SH, Wang P, Su CY, Xie ZY, Li YX, Li D, Wang S, Su HJ, Wu XH, Deng W, Wu YF, Shen HY. Bone marrow mesenchymal stem cels derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):13-19.
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Objective To investigate the clinical effect of repairing soft tissue defects of foot and ankle by sural neurovascular adipofascial flap.Methods Application of sural neurovascular adipofascial flap to repaire soft tissue defects of foot and ankle in 19 patients,inclouding 3 csaes with the soft tissue defects of medial malleolus and 16 cases of dorsum pedis from May,2006 to May,2013.The size of soft tissue defects was 3 cm ×3 cm-7 cm ×9 cm.The skin of the donor site was sutured directly.The recipient area was reshaped granulation and underwent free skin graft after the adipofascial flap survived.Results After followed up of 3-18 months,the adipofascial flaps were all survived,the recipient area was almost flat with the surrounding tissue,the donor area only leaving linear scar,the appearance and function of donor area and adopt area were satisfactory.The sensation of lateral dorsal region of foot decreased to S3,and the sensation of recipient site recovered to S2.There were some changes in pigmentation with the skin graft region.Conclusion Sural neurovascular adipofascial flap can be effectively repaired soft tissue defects of foot and ankle,and it can avoid irregularity of donor area and enlargement of adopt area,the appearance and function were satisfactory.