Effect of Silent Information Regulator on the Survival and Osteogenic Differentiation of Inflammatory Bone Marrow Mesenchymal Stem Cells by Regulating NF-κB Signaling Pathway

2020 ◽  
Vol 10 (1) ◽  
pp. 121-126
Author(s):  
Wenkun Lu ◽  
Tao Wang ◽  
Xunjian Gao ◽  
Fuqiang Yang ◽  
Jianjian Ge
Keyword(s):  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Promote Cell Proliferation ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells ◽  
Inflammation Group

Osteogenic differentiation of BMSCs is beneficial for osteoarthritis (OA) treatment. Silent information regulator (SIRT1) plays a role in endocrine diseases and aging-related diseases. However, the role of SIRT1 in OA has not yet been elucidated. Rat BMSCs were isolated and divided into control group, inflammation group (BMSCs were cultured with IL-6), SIRT1 group (SIRT1 agonist Resveratrol was added under the action of IL-6) followed by analysis of cell proliferation by MTT assay, Caspase 3 activity, ALP activity, expression of osteogenic genes Runx2 and OC and adipogenic differentiation gene PPARγ2 by Real time PCR, NF-κB expression by western blot and secretion of TNF-α and IL-6 by ELISA. In inflammation group, SIRT1 expression was significantly decreased, cell proliferation was significantly inhibited, and Caspase 3 activity was increased. Meanwhile, ALP activity, Runx2 and OC expression was decreased, PPARγ2 and NF-κB expression was increased, along with elevated TNF-α and IL-6 secretion compared to control (P < 0.05). Resveratrol can significantly promote the expression of SIRT1 in BMSCs of inflammation group, promote cell proliferation, decrease Caspase 3 activity, and increase Runx2 and OC expression. In addition, it decreased PPARγ2 and NF-κB expression and reduced the secretion of TNF-α and IL-6 (P < 0.05). The expression of SIRT1 was decreased in BMSCs under inflammation. SIRT1 overexpression in BMSCs under inflammation inhibits inflammation, promotes proliferation and osteogenic differentiation of BMSCs through regulating NF-κB signaling pathway.

Effect of 25-Hydroxycholesterol on the Differentiation of Bone Marrow Mesenchymal Stem Cells Through Wnt5/TGF-β Signaling Pathway

2019 ◽  
Vol 9 (11) ◽  
pp. 1583-1588
Author(s):  
Shaoting Li ◽  
Jinhe Zhou ◽  
Zhiqing Ye ◽  
Shenglin Wu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Control Group ◽  
Nodule Formation ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) can be multi-directionally differentiated and are widely used in tissue engineering. 25-hydroxycholesterol (25-HC) can induce osteogenesis and is involved in osteogenic formation. However, the role of 25-hydroxycholesterol in BMSCs is unclear. Rat BMSCs were isolated and divided into control group and 25-HC treatment (2 and 4 μM) group. Cell proliferation was detected by MTT assay. Caspase-3 and ALP activity was analyzed. Real time PCR was done to analyze Runx2, OPN, FABP4 and PPARγ2 expression. Red staining detects the calcified nodule formation. Wnt5 level was detected by western blot and TGF-β secretion was analyzed by ELISA. 25-HC treatment significantly inhibited cell proliferation, increased Caspase 3 activity, decreased ALP activity and the expression of Runx2 and OPN, increased expression of FABP4 and PPARγ2, decreased formation of calcified nodules, secretion of TGF-β and reduced expression of Wnt5 compared to control group (P < 0.05), and the above changes were significant with the increase of the concentration of 25-HC (P < 0.05). 25-hydroxycholesterol regulates the proliferation and apoptosis of BMSCs by regulating Wnt5/TGF-β signaling pathway, inhibiting the differentiation of BMSCs into osteogenic direction and promoting its adipogenic differentiation.

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.

Effect of Simvastatin on Bone Marrow Mesenchymal Stem Cells in Osteoporosis Rats

2019 ◽  
Vol 9 (9) ◽  
pp. 1311-1316
Author(s):  
Yuechuang Liang ◽  
Liang Ma ◽  
Yu Wu ◽  
Youwei Tian ◽  
Dongyue Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Lipid Lowering ◽  
Control Group ◽  
Activity Increase ◽  
Promote Cell Proliferation ◽  
Alp Activity ◽  
The Difference

Osteoporosis (OP) is a common and frequently-occurring disease in orthopedics. BMSCs play a role in OP. Simvastatin (SVA) is a commonly used lipid-lowering drug, but its role in OP remains unclear. Our study intends to assess SVA’s effect on BMSCs in osteoporosis rats. SD rats were randomly and equally divided into control group and OP group. BMSCs in control group and OP group were cultured in vitro treated with 5 μM and 10 μM SVA followed by analysis of cell proliferation by MTT assay, apoptosis activity by Caspase 3 activity, Wnt5/TGF-β signaling pathway protein expression by Western blot, ALP activity; Runx2 and OC expression by Real time PCR as well as BMP-2 and TGF-β secretion by ELISA. OP rat BMSCs showed significantly inhibited cell proliferation, increased Caspase 3 activity, decreased Wnt5, Runx2 and OC expression and ALP activity, as well as reudced BMP-2 and TGF-β secretion (P < 0.05). SVA can promote cell proliferation, inhibit Caspase 3 activity, increase Wnt5, Runx2 and OC expression and ALP activity, as well as promote BMP-2 and TGF-β secretion in OP rat BMSCs. Compared with OP group, the difference was statistically significant with more significant changes with increasing concentration (P < 0.05). Simvastatin activates Wnt5/TGF-β signaling pathway, regulates BMSCs proliferation and apoptosis and promotes their differentiation into osteogenic direction in OP rats.

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.

Long Noncoding RNA Metastasis-Associated Lung Carcinoma Transcript 1 (MALAT1) Inhibits Bone Marrow Mesenchymal Stem Cell Formation Through Regulating Smad/Runx2 Signaling Pathway

2020 ◽  
Vol 10 (5) ◽  
pp. 730-736
Author(s):  
Qingyun Pan ◽  
Biao Dong ◽  
Yong He ◽  
Xiaohui Wang
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Control Group ◽  
Alp Activity ◽  
Lncrna Malat1 ◽  
Sirna Group

Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is controlled by elaborate genetic programs. lncRNA MALAT1 plays an important role in many diseases. However, the role of lncRNA MALAT1 in BMSCs remains unclear. Isolated Rat BMSCs were cultured and randomly divided into control group, MALAT1 group and MALAT1 siRNA group, in which lncRNA MALAT1 plasmid and lncRNA MALAT1 siRNA were transfected into BMSCs followed by analysis of lncRNA MALAT1 expression by real time PCR, cell proliferation by MTT assay, Caspase 3 activity, ALP activity was analyzed, calcified nodules by alizarin red staining, expression of Smad1 and Smad7 by Western blot as well as Runx2 expression by real time PCR. In MALAT1 group, MALAT1 expression was significantly increased along with significantly inhibited cell proliferation, increased Caspase 3 activity, decreased ALP activity and calcified nodules, reduced expression of Smad1, Smad7 and Runx2 compared with control group (P < 0 05). MALAT1 expression in MALAT1 siRNA group was decreased with significantly promoted cell proliferation, decreased Caspase 3 activity, increased ALP activity and calcified nodules, as well as significantly elevated expression of Smad1, Smad7 and Runx2 compared with control group (P < 0 05). Up-regulation of lncRNA MALAT1 expression inhibits the Smad/Runx2 signaling pathway, thereby inhibiting BMSCs proliferation and osteogenesis. Down-regulation of lncRNA MALAT1 expression promotes Smad/Runx2 signaling pathway activation, inhibits BMSCs apoptosis, promotes proliferation and osteogenic differentiation.

Progranulin Reverses Decitabine-Induced Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Through Downregulation of NF-κB Signaling Pathway

2020 ◽  
Vol 10 (1) ◽  
pp. 87-92
Author(s):  
Shouping Zhang ◽  
Ying Wang ◽  
Lili Sun
Keyword(s):  
Bone Marrow ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells ◽  
Activity Decrease ◽  
Oil Red O Staining ◽  
Oil Red O

Decitabine can induce BMSCs adipogenic differentiation. Progranulin (PGRN) is a chondrogenic factor. However, the effect of Progranulin on the adipogenic differentiation of BMSCs induced by decitabine remains unclear. Rat BMSCs were isolated and divided into control group, Decitabine group, and Decitabine+PGRN group followed by analysis of survival rate of BMSCs cells by MTT assay, Caspase 3 activity, ALP activity, Runx2, OP and PPARγ2 expression by Real time PCR, lipids formation by Oil red O staining and the expression of NF-κB by Western blot. Decitabine treatment can significantly inhibit the proliferation of BMSCs, promote the increase of Caspase 3 activity, decrease ALP activity and the expression of Runx2 and OP, increase PPARγ2 expression, the ability of adipogenesis and NF-κB expression (P < 0005). Progranulin addition significantly promoted BMSCs proliferation, inhibited Caspase 3 activity, increased ALP activity and Runx2, OP expression, decreased PPARγ2 expression, adipogenic capacity and NF-κB expression, compared to Decitabine group (P < 0005). Decitabine inhibits BMSCs proliferation, promotes apoptosis, induces adipogenic differentiation, and inhibits osteogenic differentiation. Progranulin reverses the effect of defercitin on the induction of adipogenic differentiation of BMSCs by down-regulating the NF-κB signaling pathway.

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.

Progranulin Regulates the Differentiation of Bone Marrow Mesenchymal Stem Cells Under Inflammation Through Focal Adhesion Kinase (FAK)/Mitogen-Activated Protein Kinase (MAPK) Pathway

2020 ◽  
Vol 10 (2) ◽  
pp. 281-286
Author(s):  
Zirong Yang ◽  
Hangbo Qu ◽  
Hongting Jin
Keyword(s):  
Osteogenic Differentiation ◽  
Caspase 3 ◽  
Mapk Pathway ◽  
Mitogen Activated Protein Kinase ◽  
Control Group ◽  
Alp Activity ◽  
Mitogen Activated Protein ◽  
Marrow Mesenchymal Stem Cells ◽  
Group 5

Osteogenic differentiation of BMSCs is beneficial to the treatment of osteoarthritis. Progranulin (PGRN) is a chondrogenic factor. However, the role of progranulin in the differentiation of BMSCs under inflammation remains unclear. Rat BMSCs were isolated and divided into control group, inflammation group (treated with LPS), and PGRN group (5 and 10 /μM) followed by analysis of survival rate of BMSCs by MTT assay, Caspase 3 activity, ALP activity, expression of Runx2 and OP by real time PCR, level of MMP-3, TIMP-1, FAK and MAPK by Western blot and IL-6 and IL-10 secretion by ELISA. LPS treatment significantly inhibited BMSCs proliferation, increased Caspase 3 activity, decreased ALP activity, expression of Runx2 and OP, increased IL-6 secretion, decreased IL-10 secretion, increased MMP-3 expression, decreased expression of TIMP-1, FAK and p-P38 (P < 0.05). PGRN treatment on BMSCs under inflammation significantly promoted cell proliferation, decreased Caspase 3 activity, increased ALP activity, expression of Runx2 and OP, decreased IL-6 secretion, increased IL-10 secretion, decreased MMP-3 expression, and increased TIMP-1, FAK and p-P38 expression (P < 0.05) with more significant changes in the higher concentration. Under inflammation, BMSCs proliferation was inhibited, apoptosis was increased, and osteogenic differentiation was weakened. PGRN inhibits the proliferation of BMSCs and apoptosis, and promotes osteogenic differentiation by regulating FAK/MAPK pathway.

scholarly journals The mechanism of miR-889 regulates osteogenesis in human bone marrow mesenchymal stem cells

2019 ◽  
Vol 14 (1) ◽  
Author(s):  
Gang Xu ◽  
Zheng Ding ◽  
Hui-feng Shi
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Calcium Deposition ◽  
Control Group ◽  
Related Protein ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Abstract Background Bone marrow mesenchymal stem cells (BMMSCs) can be used for bone regeneration in the specified condition. Osteogenic differentiation of BMMSCs is controlled by microRNAs (miRNAs) and other factors. This study was aimed to identify the role and mechanism of miR-889 in regulating the osteogenic differentiation of BMMSCs. Methods Osteoporosis patients and normal control bone tissues were collected and used PCR techniques to identify the change of miR-889 and WNT7A. Moreover, the dynamic change of miR-889 and WNT7A during osteogenic differentiation of BMMSCs was also measured. Bioinformatic analysis was performed to identify the target genes and potential pathways of miR-889. Then, we constructed miR-889 mimic and inhibitor, ALP staining, ARS, osteoblastic-related protein, and Wnt β-catenin signaling pathway-related protein were also measured. WNT7A siRNA was also used to verify the function of miR-889. Results In the present study, we showed that miR-889 expression was upregulated in osteoporosis patients than healthy control. However, the miR-889 expression was downregulated during osteogenic differentiation. Bioinformatics analysis found that miR-889 targets 666 genes and mainly through Wnt β-catenin signaling pathway. Administrated miR-889 mimic, the ALP activity, and calcium deposition were decreased than the control group, while miR-889 inhibitor shown the opposite trend. And miR-889 could bind the 3′UTR of WNT7A. We further used WNT7A siRNA to explore the function of miR-889, and the results revealed that co-cultured with miR-889 inhibitor and WNT7A siRNA was associated with a reduction of ALP activity and calcium deposition and osteoblastic-related proteins than miR-889 inhibitor alone. Conclusion Our results revealed that miR-889 plays a negative role in inducing osteogenic differentiation of BMSCs through Wnt β-catenin signaling pathway.

Overexpression of Guanine Nucleotide Binding Protein, Alpha Stimulating (GNAS) Regulates Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in High Glucose Environment by Regulating ERK/P38 Signaling Pathway

2020 ◽  
Vol 10 (2) ◽  
pp. 259-264
Author(s):  
Wei Zhang ◽  
Yuanbo Wang ◽  
Song Jin ◽  
Hui Xin ◽  
Changxin Wang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Western Blot ◽  
Signaling Pathway ◽  
High Glucose ◽  
Caspase 3 ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) can treat osteoporosis. Whether GNAS affects BMSCs osteogenic differentiation under high glucose condition is unknown. Rat BMSCs were isolated and randomly divided into control group, high glucose group and GNAS group. The BMSCs were transfected with GNAS plasmid in high glucose environment followed by analysis of GNAS expression by Real time PCR and Western blot, BMSCs proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of calcified nodules by alizarin red staining, OC and BMP-2 expression by Real time PCR and expression of ERK/P38 signaling pathway protein by Western blot. In high glucose environment, GNAS expression was significantly decreased, cell proliferation was inhibited, Caspase 3 activity was increased, along with decreased ALP activity, calcified nodules formation and expression of OC, BMP-2, p-ERK1/2 and p-P38 (P < 0.05). GNAS plasmid transfected into high glucose environment BMSCs can significantly promote GNAS expression and cell proliferation, decrease Caspase 3 activity, increase p-ERK1/2 and p-P38 expression, ALP activity and calcified nodules formation as well as increase OC and BMP-2 expression (P < 0.05). GNAS1 expression is decreased in BMSCs cells in a high glucose environment. Overexpression of GNAS1 can inhibit the apoptosis of BMSCs by regulating the ERK/P38 signaling pathway, promote its proliferation and differentiation into osteogenic direction.

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