Effect of Mir-663 on the Differentiation of Bone Marrow Mesenchymal Stem Cells in Inflammatory Environment Through Regulating Oxidative Stress and Transforming Growth Factor β1

2019 ◽  
Vol 9 (11) ◽  
pp. 1600-1606
Author(s):  
Xiuhong Sun ◽  
Yujie Liu ◽  
Shujuan Zheng
Keyword(s):  
Oxidative Stress ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Control Group ◽  
Sod Activity ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

BMSCs are beneficial for the treatment of Osteoarthritis (OA). Mir-663 involves in various diseases. However, the role of Mir-663 in the differentiation of BMSCs in an inflammatory environment remains unclear. Rat BMSCs were isolated and divided into control group, and inflammation group which was treated with 1 μg/ml lipopolysaccharide (LPS), Mir-663 group and Mir-663 siRNA groups which was respectively transfected with Mir-663 plasmid and Mir-663 siRNA into LPS-treated BMSCs followed by analysis of the survival rate of BMSC by MTT, Caspase 3 activity, ALP activity, expression of osteogenic genes Runx2 and OP by Real time PCR, ROS content and SOD activity, TGF-β1 level by Real time PCR and ELISA, TNF- and IL-1 secretion by ELISA. LPS treatment significantly increased Mir-663 expression, inhibited cell proliferation, increased Caspase 3 activity, and inhibited ALP activity. Meanwhile it also significantly decreased Runx2 and OP expression, increased ROS content, decreased SOD activity and TGF-β1 expression as well as elevated TNF-α and IL-1β secretion compared to control (P < 0.05); Mir-663 plasmid transfection can further promote the above changes (P < 0.05); whereas, Mir-663 siRNA could reverse the above changes (P < 0.05). Mir-663 expression was increased in BMSCs in inflammatory environment. Down-regulation of Mir663 expression can regulate oxidative stress, up-regulate TGF-β1 level, inhibit the secretion of TNF-α and IL-1β, and promote the proliferation and osteogenic differentiation of BMSCs in inflammatory environment.

Effect of Long-Chain Non-Coding RNA GAS5 on Osteogenic/Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Oxidative Stress Through Targeting MiR-365

2019 ◽  
Vol 9 (12) ◽  
pp. 1751-1757
Author(s):  
Wenming Wu ◽  
Dongming Liang
Keyword(s):  
Oxidative Stress ◽  
Cell Proliferation ◽  
Real Time ◽  
Real Time Pcr ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Sod Activity ◽  
Stress Group ◽  
Alp Activity ◽  
Lncrna Gas5

Oxidative stress affects BMSCs. LncRNA GAS5 regulates cell proliferation and apoptosis. However, the effect of LncRNA GAS5 on osteogenesis/adipogenic differentiation of BMSCs under oxidative stress has not been reported. Rat BMSCs were cultured and randomly divided into 4 groups, normal control group; oxidative stress group; GAS5 siRNA group; GAS5 siRNA+ miR-365 inhibitor group followed by analysis of LncRNA GAS5 expression by Real time PCR, cell proliferation by MTT assay, Caspase3 activity, GAS5 and miR-365 targeting relationship by luciferase reporter assay, ALP activity, expression of Runx2, OP and PPAR 2 by Real time PCR, as well as ROS content and SOD activity. In oxidative stress group, GAS5 expression was significantly increased along with inhibited cell proliferation, increased Caspase3 activity, decreased ALP activity and the expression of Runx2 and OP, increased PPAR 2 expression and ROS content, and decreased SOD activity compared to control group (P < 0 05). miR-365 was the target miRNA of GAS5. GAS5 siRNA down-regulated GAS5 expression, significantly promoted cell proliferation, inhibited Caspase3 activity, increased ALP activity and Runx2 and OP expression, decreased PPAR 2 expression and ROS content, and increased SOD activity. (P < 0 05). However, GAS5 siRNA+ miR-365 inhibitor group reversed the effect of GAS5 siRNA. Oxidative stress promotes LncRNA GAS5 expression in BMSCs. LncRNA GAS5 regulates oxidative stress by targeting miR-365. Knockdown of GAS5 can promote BMSCs proliferation and osteogenic differentiation and inhibit adipogenic differentiation under oxidative stress.

Effect of αCGRP on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Through Smad/Runx2 Signaling Pathway

2020 ◽  
Vol 10 (6) ◽  
pp. 868-873
Author(s):  
Shengxiang Huang ◽  
Haibo Mei ◽  
Rongguo He ◽  
Kun Liu ◽  
Jin Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

The α-calcitonin gene-related peptide (α-CGRP) regulates bone metabolism and has potential applications in enhancing bone remodeling in vivo. However, α-CGRP's role in bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation remain unclear. Rat BMSCs were separated into control group, α-CGRP group and α-CGRP siRNA group, in which BMSCs were transfected with α-CGRP plasmid and α-CGRP siRNA respectively followed by analysis of α-CGRP level by real time PCR and ELISA, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of calcified nodules by alizarin red staining, Smad1 and Smad7 level by Western blot and Runx2 by real time PCR. αCGRP transfection into BMSCs significantly up-regulated CGRP, which could promote cell proliferation, inhibit Caspase 3 activity, promote ALP activity, increase calcified nodules formation and upregulate Smad1, Smad7 and Runx2 compared to control (P < 0.05); transfection of αCGRP siRNA significantly down-regulated CGRP in BMSCs, inhibited cell proliferation, promoted Caspase 3 activity, inhibited ALP activity, inhibited calcified nodules formation and downregulate Smad1, Smad7 and Runx2 (P < 0.05). αCGRP overexpression promotes the Smad/Runx2 signaling, which in turn promotes BMSCs proliferation and osteogenesis. Decreased αCGRP level inhibits Smad/Runx2 signaling, promotes BMSCs apoptosis, inhibits proliferation and osteogenic differentiation.

The Regulatory Effect of Mir-149 on Bone Marrow Mesenchymal Stem Cells in Osteoporosis Rats and Its Related Mechanisms

2019 ◽  
Vol 9 (8) ◽  
pp. 1127-1132 ◽  
Author(s):  
Long Wang ◽  
Jian Yu
Keyword(s):  
Cell Proliferation ◽  
Real Time ◽  
Real Time Pcr ◽  
Mapk Signaling ◽  
Control Group ◽  
Regulatory Effect ◽  
Erk Mapk ◽  
Alp Activity ◽  
Sd Rats ◽  
Marrow Mesenchymal Stem Cells

BMSCs play an important role in osteoporosis (OP) and their differentiation can lead to OP progression. Mir-149 can participate in the regulation of BMSCs. However, the effect of Mir-149 on BMSCs in osteoporosis remains unclear. SD rats were divided control group and OP group. OP rat BMSCs were transfected with Mir-149 siRNA followed by analysis of Mir-149 expression by real time PCR, cell proliferation by MTT assay, apoptosis by flow cytometry, ERK/MAPK signaling protein expression by western blot, ALP activity, as well as expression of osteogenic genes Runx2 and OC by real time PCR. Mir-149 expression was significantly increased in BMSCs of OP rats, cell proliferation was inhibited, apoptosis was increased, as well as p-ERK1/2 expression, ALP activity and expression of Runx2 and OC was decreased compared to control group (P < 0.05). Transfection of Mir-149 siRNA into OP rat BMSCs reduced Mir-149 expression, promoted cell proliferation, decreased apoptotic rate, increased p-ERK1/2 expression, ALP activity and Runx2 and OC expression. Compared with OP group, the differences were statistically significant (P< 0.05). Mir-149 expression was increased in OP rat BMSCs. Down-regulation of Mir-149 promoted the activation of ERK/MAPK signaling pathway, inhibited apoptosis of BMSCs and promoted the proliferation and osteogenic differentiation of BMSCs in OP rats.

Effect of LncRNA DGCR5 on Bone Marrow Mesenchymal Stem Cells of Osteoporosis Rats Through Regulation of ERK/P38 Signaling Pathway

2019 ◽  
Vol 9 (11) ◽  
pp. 1589-1594
Author(s):  
Xu Tong ◽  
Renjian Zheng ◽  
Linjing Shu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Signaling Pathway ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Control Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation plays an important role in Osteoporosis (OP). LncRNA DGCR5 participates in OP development. However, LncRNA DGCR5's effect on BMSCs in osteoporosis rats and related mechanisms have not been elucidated. SD rats were divided into control group and OP group. Rat BMSCs were cultured and transfected with LncRNA DGCR5 siRNA followed by analysis of LncRNA DGCR5 expression by Real time PCR, cell proliferation by MTT assay, Caspase 3 activity, of ERK/P38 signaling pathway protein expression by Western blot, ALP activity, and the osteogenic genes Runx2 and OC expression by Real time PCR. LncRNA DGCR51 expression was increased in BMSCs of OP rats. Compared with control group, cell proliferation was significantly inhibited, Caspase 3 activity was increased, p-ERK1/2 and p-P38 were downregulated, ALP activity, Runx2 and OC expression was decreased (P < 0.05). DGCR51 siRNA transfection into OP rat BMSCs significantly reduced DGCR51 expression, promoted cell proliferation, decreased Caspase 3 activity, increased p-ERK1/2 and p-P38 expression, increased ALP activity, Runx2 and OC expression compared to OP group (P < 0.05). LncRNA DGCR51 expression is increased in OP rat BMSCs. Down-regulation of LncRNA DGCR51 promoted the activation of ERK/P38 signaling pathway, thereby inhibiting the apoptosis of BMSCs and promoting proliferation and osteogenic differentiation of BMSC in OP rats.

Effects of LncRNA SNHG15 on Osteogenic/Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Oxidative Stress by TGFβ/Smad Signaling Pathway

2019 ◽  
Vol 9 (9) ◽  
pp. 1273-1278
Author(s):  
Chengqiang Zhang ◽  
Lihua Fang ◽  
Xiaoping Liu ◽  
Rui Li ◽  
Tingting Nie ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Bone Marrow ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Ros Production ◽  
Sod Activity ◽  
Smad Signaling ◽  
Alp Activity ◽  
Smad Signaling Pathway ◽  
Marrow Mesenchymal Stem Cells

Oxidative stress can affect bone marrow mesenchymal stem cells (BMSCs). LncRNA SNHG15 involves in a variety of cellular physiological and pathological processes. However, LncRNA SNHG15’s effect on osteogenesis/adipogenic differentiation of BMSCs under oxidative stress remains unclear. BMSCs were divided into normal control group; oxidative stress group; SNHG15 group in which SNHG15 plasmid was transfected under oxidative stress condition followed by analysis of the expression of LncRNA SNHG15, RUNX2 and OPN by real-time quantitative PCR, reactive oxygen species (ROS) and superoxide dismutase (SOD) activity, FABP4 and PPARγ2 mRNA expression, alkaline phosphatase (ALP) activity and TGFβ/Smad signaling by Western blot. Under oxidative stress, the expression of SHHG15, RUNX2 and OPN mRNA was significantly decreased, ROS production was increased, SOD activity was reduced, along with increased expression of FABP4 and PPARγ2 protein, decreased ALP activity, as well as reduced expression of TGFβ1, Smad2, and Smad7 compared to control group (P < 0.05). Transfection of LncRNA SNHG15 plasmid significantly up-regulated the expression of SNHG15, RUNX2 and OPN mRNA, decreased ROS production, increased SOD activity, decreased expression of FABP4 and PPARγ2 mRNA, and increased ALP activity as well as increased expression of TGFβ1, Smad2, and Smad7 compared to control group (P < 0.05). The expression of SNHG15 is decreased in BMSCs under oxidative stress. Up-regulation of SNHG15 expression can improve the redox balance through TGFβ/Smad signaling pathway, promote osteogenic differentiation of BMSCs under oxidative stress, and inhibit its differentiation into fat.

Overexpression of Heme Oxygenase-1 Promotes Osteoblast Differentiation of Osteoporosis Rat Bone Marrow Stromal Cells in Bone Morphogenetic Protein-Dependent Manner

2019 ◽  
Vol 9 (11) ◽  
pp. 1614-1620
Author(s):  
Jiangrong Fan ◽  
Yong Zheng ◽  
Jingyang You
Keyword(s):  
Cell Proliferation ◽  
Real Time ◽  
Heme Oxygenase ◽  
Real Time Pcr ◽  
Osteoblast Differentiation ◽  
Caspase 3 ◽  
Control Group ◽  
Heme Oxygenase 1 ◽  
Dependent Manner ◽  
Alp Activity

BMSCs play a role in osteoporosis (OP) and their differentiation can lead to OP progression. Heme oxygenase-1 (HO-1) involves in many diseases, but the effect of HO-1 on osteoblast differentiation of BMSCs in OP rats remains unclear. SD rats were divided into control group and OP group. Rats BMSCs in OP group were cultured in vitro, HO-1 expression was up-regulated by HO-1 agonist hemin, and BMPR inhibitor LDN-19318 was added followed by analysis of HO-1 expression by real time PCR and ELISA, cell proliferation by MTT assay, apoptosis by Caspase 3 activity, BMP-2 expression by Western blot, ALP activity, expression of Runx2 and OC by real time PCR. In OP group, HO-1 expression was significantly decreased, cell proliferation was inhibited, Caspase 3 activity was increased along with decreased ALP activity and expression of Runx2, OC and BMP-2 compared to control (P < 0.05). Up-regulation of HO-1 expression significantly promoted cell proliferation, reduced Caspase 3 activity, increased ALP activity, and expression of Runx2, OC and BMP-2 (P < 0.05). However, inhibition of HO-1 significantly promoted bone differentiation after the addition of BMPR inhibitor LDN-193189 (P < 0.05). HO-1 expression is decreased in BMSCs of OP group rats. Up-regulation of HO-1 promoted BMSCs proliferation in OP rats in BMP-dependent manner, inhibited apoptosis, and promoted osteoblast differentiation.

Effect of Chordin-Like 1 on Differentiation of Bone Marrow Mesenchymal Stem Cells Through Wnt5/TGF-β Signaling Pathway

2019 ◽  
Vol 9 (9) ◽  
pp. 1304-1310
Author(s):  
Qing Yang ◽  
Lei Wu ◽  
Yang Liu ◽  
Bing Yuan
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Control Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Chordin-like 1 (CHRDL1) functions in multiple tissues and organs. However, whether CHRDL1 affects bone marrow mesenchymal stem cells (BMSCs) differentiation remain unclear. Rat BMSCs were isolated and divided into control group, CHRDL1 group and CHRDL1 siRNA group followed by analysis of CHRDL1 level by real time PCR and ELISA, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, expression of o Runx2, OC and PPARγ2 by Real time PCR, TGF-β secretion by ELIS, and Wnt5 protein expression by Western blot. CHRDL1 expression was significantly increased in CHRDL1 group, along with significantly promoted cell proliferation, decreased Caspase 3 activity, increased ALP activity and expression of Runx2 and OC, decreased PPARγ2 expression, increased TGF-β secretion and Wnt5 expression compared to control group (P < 0.05). However, CHRDL1 siRNA transfection significantly decreased CHRDL1 expression, inhibited cell proliferation, increased Caspase 3 activity, decreased ALP activity and Runx2 and OC expression, increased PPARγ2 expression, decreased TGF-β secretion and Wnt5 expression. (P < 0.05). Down-regulation of CHRDL1 expression in BMSCs promotes Wnt5/TGF-β signaling transduction, which in turn increases BMSCs proliferation and osteogenic differentiation. Up-regulation of CHRDL1 expression in BMSCs inhibited the activation of Wnt5/TGF-β signaling pathway, promoted BMSCs apoptosis, and inhibited BMSCs proliferation and osteogenic differentiation.

Down-Regulation of Notch-2 Promotes the Osteogenic Differentiation of Bone Marrow Stromal Cells in High Glucose Environment by Targeting Fat Mass and Obesity-Associated Protein

2019 ◽  
Vol 9 (10) ◽  
pp. 1459-1465
Author(s):  
Zhenhua Zhang ◽  
Dianguo Li ◽  
Xiaogang Sun ◽  
Jialong Xu
Keyword(s):  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
High Glucose ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Down Regulation ◽  
Sod Activity ◽  
Alp Activity ◽  
High Glucose Group

BMSCs Differentiation into osteoblasts is beneficial for treating osteoporosis. Notch signaling pathway regulates the proliferation of BMSCs. However, Notch-2's effect on osteogenic differentiation of BMSCs in high glucose environment remains unclear. Rat BMSCs were isolated and divided into control group, high glucose group, Notch-2 siRNA group, in which BMSCs cells were transfected with Notch-2 siRNA under high glucose environment followed by analysis of Notch-2 mRNA expression by Real time PCR, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, expression of osteogenic genes Runx2 and BMP-2 by Real time PCR, TNF-α and IL-2 secretion by ELISA, myeloperoxidase (MPO) and superoxide dismutase (SOD) activity and FTO protein expression by Western blot. In high glucose group, Notch-2 expression was increased with inhibited cell proliferation, increased Caspase 3 activity and decreased ALP activity. Meanwhile, high glucose significantly increased MPO content, decreased SOD activity, increased TNF-α and IL-2 secretion and FTO expression, and reduced Runx2 and BMP-2 expression (P < 0.05); Notch-2 siRNA transfection significantly down-regulated Notch-2 expression, promoted BMSCs cell proliferation, decreased Caspase 3 activity and increased ALP activity. In addition, it also significantly decreased MPO content, increased SOD activity, decreased TNF-α and IL-2 and FTO expression as well as increased Runx2 and BMP-2 expression (P < 0.05). Notch-2 is upregulated in BMSCs under high glucose. Down-regulation of Notch-2 in BMSCs under high glucose environment could promote BMSCs proliferation and osteogenic differentiation.

Osteoprotegerin Protects H2O2-Induced Bone Marrow Mesenchymal Stem Cells Damage

2019 ◽  
Vol 9 (8) ◽  
pp. 1154-1159
Author(s):  
Leitao Qi ◽  
Hongqing An
Keyword(s):  
Oxidative Stress ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Caspase 3 ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Stress Model ◽  
Sod Activity ◽  
Marrow Mesenchymal Stem Cells

Oxidative stress causes damage and apoptosis of bone marrow mesenchymal stem cells (BMSCs). Osteoprotegerin (OPG) regulates bone differentiation. However, whether OPG plays a role in osteogenic/adipogenic differentiation under oxidative stress remains poorly understood. Rat BMSCs were randomly divided into control group; H2O2 group, in which H2O2 was used to prepare oxidative stress model; OPG group, in which 5 μM OPG was added to BMSCs based on oxidative stress model followed by analysis of BMSCs proliferation by MTT assay, BMSCs apoptosis, expression of RUNX2, OPN, FABP4 and PPARγ2 by real time PCR, OPG and RANKL protein expression by Western blot as well as SOD activity and ROS content by spectrophotometry. Compared with control group, H2O2 significantly decreased BMSCs proliferation, increased Caspase 3 activity, downregulated RUNX2 and OPN, upregulated FABP4 and PPARγ2, decreased SOD activity and increased ROS content (P < 0.05). Under H2O2 treatment, OPG addition significantly promoted BMSCs proliferation, decreased Caspase 3 activity, increased RUNX2 and OPN expression, decreased FABP4 and PPARγ2 expression, increased SOD activity, and decreased ROS content and RANKL expression (P < 0.05). OPG can promote osteogenic differentiation of BMSCs under oxidative stress, inhibit adipogenic differentiation, and protect BMSCs from damage and promote BMSCs proliferation by regulating OPG/RANKL signaling pathway.

Increased Expression of TXNIP Facilitates Oxidative Stress in Nasal Epithelial Cells of Patients With Chronic Rhinosinusitis With Nasal Polyps

2020 ◽  
pp. 194589242098241
Author(s):  
Hai Lin ◽  
Guangyi Ba ◽  
Ru Tang ◽  
Mingxian Li ◽  
Zhipeng Li ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Western Blotting ◽  
Real Time Pcr ◽  
Nasal Polyps ◽  
Sod Activity ◽  
Nasal Tissue ◽  
Sod Assay

Background Oxidative stress plays crucial roles in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). Thioredoxin-interacting protein (TXNIP) is essential in the process of triggering oxidative stress. However, its role and mechanism in CRSwNP remain unclear. The present study sought to explore the role and mechanism of TXNIP in the pathogenesis of CRSwNP. Methods Western blotting, real-time PCR and immunohistochemistry (IHC) were employed to assess TXNIP, thioredoxin (TRX) expression in nasal tissue samples from patients with CRSwNP and control subjects. MDA level and SOD activity in nasal tissue homogenates were measured using MDA and SOD Assay Kit. To evaluate the role and mechanism of TXNIP in CRSwNP, human nasal epithelial cells (HNECs) were cultured and stimulated using TXNIP siRNA, with or without N-acetylcysteine (NAC, an ROS scavenger). Western blotting, real-time PCR, ROS detecting dye DCFH-DA, MDA and SOD Assay Kit were performed to assess the effects and mechanisms of stimulators on the cells. Results We found significantly increased levels of TXNIP and decreased levels of TRX protein, mRNA, positive cells, increased MDA level and decreased SOD activity in CRSwNP patients compared with control subjects. In vitro study, significantly altered levels of TXNIP, TRX, MDA, SOD and ROS in HNECs were found following treatment of TXNIP siRNA with or without NAC on HNECs. Conclusion TXNIP expression was increased and TRX expression was decreased in CRSwNP at both protein and mRNA levels. MDA levels were increased and SOD activities were decreased in CRSwNP. TXNIP may have negative association with TRX, and then decrease SOD activities and increase MDA levels, resulting in the upregulation of ROS and oxidative stress in HNECs, which may play a pivotal role in the pathogenesis of CRSwNP. Future studies are expected to further explore the role and mechanism of TXNIP in CRSwNP.

Sign in / Sign up

Export Citation Format

Share Document