Effect of miR-9 on Chondrocyte Apoptosis in Rats with Osteoarthritis by Regulating Notch1

2020 ◽  
Vol 10 (3) ◽  
pp. 371-378
Author(s):  
Xiao Zhong ◽  
Xiaodong Deng ◽  
Jian Luo ◽  
Ming Xiong ◽  
Wen Li ◽  
...  
Keyword(s):  
Western Blot ◽  
Caspase 3 ◽  
Notch Signaling Pathway ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Chondrocyte Apoptosis ◽  
Control Group ◽  
Matrix Degradation ◽  
Qrt Pcr ◽  
Protein Expressions

Objective: Notch1 is an important receptor in Notch signaling pathway. Abnormal Notch1 expression or function is associated with the pathogenesis of OA. There is evidence that the abnormal expression of miR-9 is related to the pathogenesis of OA. Bioinformatics analysis showed that there was a targeting relationship between miR-9 and Notch1’s 3′-UTR, suggesting that there might be a mutual regulation between them. In this study, we established an OA rat model to investigate whether miR-9 plays a role in regulating Notch1 expression, affecting cartilage matrix degradation and chondrocyte apoptosis in OA. Methods: OA model rats were divided into three groups: OA group, OA + agomir group, OA + miR-9 agomir group. The content of Hyp in articular fluid was measured by ELISA, the activity of caspase-3 was detected by kit, the mRNA expressions of miR-9, COL2A1, Notch1 and Hes5 was detected by qRT-PCR, and the protein expressions of Notch1 and Hes5 in articular cartilage was detected by Western blot. Chondrocytes were separated into control group, IL-1β +miR-NC group, IL-1β +miR-9 mimic group followed by analysis of mRNA expressions of miR-9, COL2A1, Notch1 and Hes5 were detected by qRT-PCR, protein expressions of Notch1 and Hes5 by Western blot, and cell apoptosis by flow cytometry. Results: Compared with Sham group, the levels of IL-1β , Hyp, Notch1, Hes5 and Caspase-3 in the synovial fluid of rats in OA group were increased significantly, while the mRNA expressions of miR-9, COL2A1 and Notch1 and Hes5 level in cartilage tissue were decreased significantly. Intra-articular injection of miR-9 agomir could significantly reduce the content of Hyp, the mRNA expressions of Notch1 and Hes5 and the activity of caspase-3 in cartilage tissue of OA rats, increase the mRNA expressions of miR-9 and COL2A1, and decrease the protein expressions of Notch1 and Hes5. After treated with IL-1β , the expressions of Notch1 and Hes5 were increased, the mRNA expressions of miR-9 and COL2A1 were decreased, and apoptosis of chondrocyte was increased in chondrocyte. After miR-9 mimic transfection, miR-9 and COL2A1 mRNA expressions were increased, Notch1 and Hes5 expressions were decreased, and apoptosis of chondrocyte was decreased under IL-1β treatment. Conclusion: Reduced miR-9 expression upregulates Notch1 expression and promotes the pathogenesis of OA. Increased miR-9 expression can inhibit Notch1 expression, reduce cartilage matrix degradation and inhibit chondrocyte apoptosis.

Celecoxib Attenuates Cartilage Matrix Damage in Arthritis Rats by Inhibiting NF-κ B

2020 ◽  
Vol 10 (4) ◽  
pp. 531-537
Author(s):  
Lukuan Du ◽  
Zhenghui Jiang ◽  
Zhaohui Wang ◽  
Liming Wang
Keyword(s):  
Caspase 3 ◽  
Sham Group ◽  
Culture Supernatant ◽  
Cartilage Tissue ◽  
Chondrocyte Apoptosis ◽  
Control Group ◽  
Therapeutic Effects ◽  
Protein Expressions ◽  
Col2a1 Expression ◽  
Matrix Damage

Objective: Celecoxib selectively inhibits the activity of COX-2 and the production of prostaglandin (PG), and plays a therapeutic role in treating osteoarthritis (OA). NF-κB signaling and IL-1α and TNFα are involved in OA pathogenesis. This study explored whether Celecoxib might exert therapeutic effects on OA through regulating NF-κB signaling and IL-1β and TNF release in OA rat model. Method : The contents of MMP-13, Hyp, IL-1β and TNFα in synovial fluid were detected by ELISA. The protein expressions of NF-κ B p-p65, COL2A1 and the activity of caspase-3 were detected. OA model rats were separated into OA group and OA+ Celecoxib group followed by analysis of MMP-13, Hyp, IL-1β and TNF level in articular fluid by ELISA and p-p65 and COL2A1 level and caspase-3 activity by western blot. Rat cartilage tissue was cultured and divided into control group, LPS group and LPS+ Celecoxib group followed by analysis of expressions of p-p65 and COL2A1 in cartilage tissue, IL-1 and TNF content in culture supernatant, and chondrocyte apoptosis. Results: Compared with Sham group, p-p65 expression and caspase-3 activity in cartilage tissue of OA rats was increased and COL2A1 level was reduced. Meanwhile the expression of MMP-13, Hyp, IL-1β and TNF in articular fluid of OA rats was increased. Compared to OA group, p-p65 expression and caspase-3 activity was decreased and COL2A1 expression was increased in OA+ Celecoxib treatment group along with decreased MMP-13, Hyp, IL-1β and TNF level in articular fluid. p-p65 expression and caspase-3 activity in LPS group was increased and COL2A1 expression was decreased with increased IL-beta; and TNF content. Compared to LPS group, p-p65 expression and caspase-3 activity was decreased and COL2A1 expression was increased in LPS+ Celecoxib group with decreased content of IL-1β and TNFα. Conclusion: Celecoxib can protect cartilage in OA by inhibiting NF-κB activation and IL-1β and TNF release, and decreasing cell apoptosis in inflammatory environment.

miR-211 Reduces the Degradation of Articular Cartilage Matrix by Targeting Matrix Metalloproteninase 9

2019 ◽  
Vol 9 (9) ◽  
pp. 1292-1297
Author(s):  
Yumei Tian ◽  
Jian Wang
Keyword(s):  
Cartilage Damage ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Control Group ◽  
Matrix Degradation ◽  
Mankin Score ◽  
Joint Cavity ◽  
Mmp9 Expression ◽  
Col2a1 Expression ◽  
And Cartilage

Matrix metalloproteninase 9 (MMP9) promotes osteoarthritis (OA) cartilage matrix degradation. Abnormal miR-211 expression is associated with OA. Bioinformatics analysis showed a targeted relationship between miR-211 and MMP9 mRNA 3′-UTR. Our study intends to evaluate miR-211’s role in the destruction of chondrocyte matrix in OA model rats. The OA model rats were divided into OA group, OA group+agomir-NC group, OA group+agomir-211 group, followed by analysis of the arthritis Mankin score, Hyp and IL-1β content by ELISA, MMP-9 and COL2A1 expression by western blot and miR-211 level by qRT-PCR. The cartilage tissues were divided into control group, IL-1β+ agomir-NC group, IL-1β+ agomir-211 group, and the expression of MMP-9 and COL2A1 in chondrocytes were detected. Compared with Sham group, IL-1β and Hyp levels in the joint cavity fluid of the OA group were significantly increased and miR-211 expression was significantly decreased with increased MMP9 expression and decreased COL2A1 expression. Injection of agomir-211 into the joint cavity of OA rats significantly reduced MMP9 expression in cartilage tissue, decreased Hyp content in the joint cavity fluid, increased COL2A1 expression, and decreased Mankin score and cartilage damage. IL-1β treatment significantly up-regulated MMP9 expression and decreased COL2A1 expression; miR-211 overexpression attenuated IL-1β-induced cartilage matrix degradation and cartilage destruction. miR-211 plays a role in regulating MMP9 expression and promoting OA. miR-211 overexpression inhibits MMP9 expression, reduces the degradation of cartilage matrix and increases collagen synthesis, thus ameliorating OA development.

miR-155 Regulates GSK-3β Expression and Affects Cartilage Matrix Degradation in the Pathogenesis of Osteoarthritis

2019 ◽  
Vol 9 (8) ◽  
pp. 1133-1139
Author(s):  
Xianjun Chen ◽  
Lin Shi ◽  
Qingjiang Pang ◽  
Xiao Yu ◽  
Ji Yang
Keyword(s):  
Caspase 3 ◽  
Sham Group ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Control Group ◽  
Traumatic Amputation ◽  
Normal Cartilage ◽  
Pathway Activity ◽  
Col2a1 Expression ◽  
Gsk 3Β

Abnormal expression of GSK-3β is associated with the pathogenesis of osteoarthritis (OA). The miR-155 expression in cartilage tissue of OA patients was significantly elevated. A relationship between miR-155 and GSK-3β is predicted by the software. This study investigated whether miR155 regulates GSK-3β expression, affects Wnt/β-catenin pathway activity in OA. The cartilage tissues of OA patients and normal cartilage tissues after traumatic amputation were collected. The OA rat model was established and divided into OA + antagomir control group, OA + miR-155 antagomir group followed by analysis of expression of miR-155, GSK-3β, β-catenin and COL2A1 by qRT-PCR, Hyp content by ELISA, caspase-3 activity, and GSK-3β, β-catenin, COL2A1 expression by western blot. Compared to control group, the expression of miR-155 and -catenin in cartilage tissue of OA patients was significantly elevated and GSK-3β level was reduced. There was a targeted regulation relationship between miR-155 and GSK-3β. OA group had significantly higher miR-155 and β-catenin expression and lower GSK-3 and COL2A1 level in the cartilage than sham group. Compared with OA + antagomir control group, miR-155 and β-catenin expression and caspase-3 activity in OA + miR-155 antagomir group were significantly decreased, with increased expression of GSK3β and COL2A1 and reduced Hyp content. Increased miR-155 expression can decrease GSK-3β expression, enhance Wnt/β-catenin pathway activity, promote the degradation and destruction of cartilage matrix and inhibit miR-155 expression, thus ameliorating the development and progress of OA.

scholarly journals Effect of Sulfated Polysaccharide from Undaria pinnatifida (SPUP) on Proliferation, Migration, and Apoptosis of Human Prostatic Cancer

2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Xiaolin Xu ◽  
Xin Zhu ◽  
Wenglong Lu ◽  
Yandong He ◽  
Yihan Wang ◽  
...  
Keyword(s):  
Western Blot ◽  
Cancer Cells ◽  
Prostatic Cancer ◽  
Caspase 3 ◽  
Undaria Pinnatifida ◽  
Sulfated Polysaccharide ◽  
Control Group ◽  
Caspase 9 ◽  
Qrt Pcr ◽  
Du145 Cells

Objective. To observe the effect of sulfated polysaccharide from Undaria pinnatifida (SPUP) on proliferation, migration, and apoptosis of human prostatic cancer. Methods. DU145 human prostate cancer cells were cultured in vitro, and the proliferation activity both in the control group and the SPUP treatment groups (25, 50, 100, 200 μg/ml) was measured by CCK-8 assay. The wound healing assay was conducted to detect the cell migration. Cell apoptosis was measured by flow cytometry. The protein and mRNA expressions of matrix metalloproteinase-9 (MMP-9) and apoptosis-related factor Bax were detected by qRT-PCR and Western blot. The expressions of cleaved caspase-3 and cleaved caspase-9 were also determined by Western blot. Results. (1) CCK-8 results showed that the proliferative activity of DU145 cells was significantly decreased with the increase of SPUP treatment concentration (P<0.05) in a dose-dependent manner and that the inhibitory effect of SPUP was most significant at 72 h (P<0.05) as compared with the control group; (2) the migration rate of SPUP-treated cells was significantly decreased (P<0.05) as compared with the control group. And the results of qRT-PCR and Western blot assays showed that SPUP inhibited the expression of MMP-9 in DU145 cells; (3) compared with the control group, the SPUP-treated groups had increased apoptosis of the cells. The expressions of apoptosis-related factors cleaved caspase-3, cleaved caspase-9, and Bax were upregulated (P<0.05), and the mRNA expression of Bax was increased (P<0.05). Conclusion. SPUP showed an antitumor activity in prostatic cancer, and the underlying mechanism may be pertaining to inhibition of migration, proliferation, and induction of apoptosis of cancer cells.

scholarly journals miR-146a-5p Promotes Chondrocyte Apoptosis and Inhibits Autophagy of Osteoarthritis by Targeting NUMB

Cartilage ◽  
2021 ◽  
pp. 194760352110235
Author(s):  
Hongjun Zhang ◽  
Wendi Zheng ◽  
Du Li ◽  
Jia Zheng
Keyword(s):  
Caspase 3 ◽  
Cartilage Tissue ◽  
Luciferase Reporter ◽  
Chondrocyte Apoptosis ◽  
Knee Cartilage ◽  
Before And After ◽  
Related Proteins ◽  
Human And Mouse

Objective miR-146a-5p was found to be significantly upregulated in cartilage tissue of patients with osteoarthritis (OA). NUMB was shown to be involved in the autophagy regulation process of cells. We aimed to learn whether NUMB was involved in the apoptosis or autophagy process of chondrocytes in OA and related with miR-146a-5p. Methods QRT-PCR was used to detect miR-146a-5p level in 22 OA cartilage tissues and 22 controls. The targets of miR-146a-5p were analyzed using software and the luciferase reporter experiment. The apoptosis and autophagy, and related proteins were detected in chondrocytes treated with miR-146a-5p mimic/inhibitor or pcDNA3.1-NUMB/si-NUMB and IL-1β, respectively. In vivo experiment, intra-articular injection of miR-146a-5p antagomir/NC was administered at the knee of OA male mice before and after model construction. Chondrocyte apoptosis and the expression of apoptosis and autophagy-related proteins were also detected. Results miR-146a-5p was highly expressed in knee cartilage tissue of patients with OA, while NUMB was lowly expressed and negatively regulated by miR-146a-5p. Upregulation of miR-146a-5p can promote cell apoptosis and reduce autophagy of human and mouse chondrocytes by modulating the levels of cleaved caspase-3, cleaved PARP, Bax, Beclin 1, ATG5, p62, LC3-I, and LC3-II. Increasing the low level of NUMB reversed the effects of miR-146a-5p on chondrocyte apoptosis and autophagy. Intra-articular injection of miR-146a-5p antagomir can also reverse the effects of miR-146a-5p on the apoptosis and autophagy of knee joint chondrocytes in OA mice. Conclusion Downregulation of miR-146a-5p suppresses the apoptosis and promotes autophagy of chondrocytes by targeting NUMB in vivo and in vitro.

scholarly journals Silica nanoparticles inducing the apoptosis of spermatocyte cell through microRNA-450b-3p targeting MTCH2-mediating mitochondrial signaling pathway

2020 ◽  
Author(s):  
Guiqing Zhou ◽  
Jianhui Liu ◽  
Xiangyang Li ◽  
Yujian Sang ◽  
Yue Zhang ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Silica Nanoparticles ◽  
Caspase 3 ◽  
Reproductive Toxicity ◽  
Control Group ◽  
Caspase 9 ◽  
Protein Expressions ◽  
Underlying Mechanisms

Abstract Background: Silica nanoparticles (SiNPs) are found in environmental particulate matter and are proven to have adverse effects on fertility. The relationship and underlying mechanisms between miRNAs and apoptosis induced by SiNPs during spermatogenesis is currently ambiguous. Experimental design: The present study was designed to investigate the role of miRNA-450b-3p in the reproductive toxicity caused by SiNPs. In vivo, 40 male mice were randomly divided into control and SiNPs groups, 20 per group. The mice in the SiNPs group were administrated 20 mg/kg SiNPs by tracheal perfusion once every 5 days, for 35 days, and the control group were given the equivalent of a normal luminal saline. In vitro, spermatocyte cells were divided into 0 and 5 μg/mL SiNPs groups, after passaged for 30 generations, the GC-2spd cells in 5 μg/mL SiNPs groups were transfected with miRNA-450b-3p and its mimic and inhibitor. Results: In vivo, the results showed that SiNPs damaged tissue structures of testis, decreased the quantity and quality of the sperm, reduced the expression of miR-450b-3p, and increased the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, and Caspase-3 in the testis. In vitro, SiNPs obviously repressed the viability and increased the LDH level and apoptosis rate, decreased the levels of the miR-450b-3p, significantly enhanced the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, Caspase-3; while the mimic of miR-450b-3p reversed the changes induced by SiNPs, but inhibitor further promoted the effects induced by SiNPs.Conclusion: The result suggested that SiNPs could induce the spermatocyte apoptosis by inhibiting the miR-450b-3p expression to target promoting the MTCH2 resulting in activating mitochondrial apoptotic signaling pathways in the spermatocyte cells.

Resveratrol Reduces Cartilage Matrix Degradation and Affects Chondrocyte Apoptosis in Rats with Traumatic Arthritis

2020 ◽  
Vol 10 (2) ◽  
pp. 239-245
Author(s):  
Zhendong Liu ◽  
Kongbin Zhang ◽  
Yuesong Weng ◽  
Jianqiang Huang ◽  
Lu You
Keyword(s):  
Protein Expression ◽  
Protein Level ◽  
Cell Apoptosis ◽  
Cartilage Tissue ◽  
Chondrocyte Apoptosis ◽  
Therapeutic Effects ◽  
Matrix Degradation ◽  
Mankin Score ◽  
Joint Cavity ◽  
Traumatic Arthritis

Objective: Resveratrol (Resv) is a polyphenolic compound with anti-inflammatory, anti-oxidant, and other pharmacological effects in rheumatoid arthritis (RA). However, whether Resv has protective and therapeutic effects in post-traumatic arthritis (PTA) remains poorly understood. We explored the function of Resv in the treatment of PTA and reducing chondrocyte matrix degradation and chondrocyte apoptosis in PTA rats. Methods: The PTA rat model was established and divided into sham group and Resv gavage group. MMP-13 and Hyp contents in the joint cavity fluid were tested by ELISA. MMP-13 and COL2A1 protein expressions in the articular cartilage tissue were detected by Western blot. Mankin score was measured. The chondrocytes cultured in vitro and divided into control group, IL-1β group, and IL-1β + Resv group. Flow cytometry was adopted to detect cell apoptosis. Results: MMP-13 and Hyp contents were significantly increased, MMP-13 protein expression was significantly upregulated, COL2A1 protein expression was significantly reduced, while Mankin score was increased in PTA group in comparison to those in normal rats. Resv treatment significantly reduced MMP-13 protein expression and elevated COL2A1 protein level in cartilage tissue, decreased MMP-13 and Hyp contents in joint cavity fluid, and reduced Mankin score. After the treatment of IL-1β, the level of MMP-13 protein was significantly elevated, with decreased COL2A1 protein level in chondrocytes, and enhanced chondrocytes apoptosis. Resv intervention significantly inhibited MMP-13 protein and increased COL2A1 level in chondrocytes, and attenuated cell apoptosis. Conclusion: Resv inhibits MMP-13 expression, attenuates chondrocyte matrix degradation and apoptosis, indicating it plays a therapeutic value in treating PTA.

scholarly journals Sanmiao formula inhibits chondrocyte apoptosis and cartilage matrix degradation in a rat model of osteoarthritis

2014 ◽  
Vol 8 (4) ◽  
pp. 1065-1074 ◽  
Author(s):  
YING XU ◽  
GUO-JING DAI ◽  
QIAN LIU ◽  
ZHEN-LI LIU ◽  
ZHI-QIAN SONG ◽  
...  
Keyword(s):  
Rat Model ◽  
Cartilage Matrix ◽  
Chondrocyte Apoptosis ◽  
Matrix Degradation ◽  
And Cartilage

scholarly journals Integrin α7 knockdown suppresses cell proliferation, migration, invasion and endothelium-mesenchymal transformation in hepatocellular carcinoma

2019 ◽  
Author(s):  
Zhiyong Wu ◽  
Xiaoyu Kong ◽  
Zhihui Wang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Proliferation ◽  
Western Blot ◽  
Cell Counting ◽  
Control Group ◽  
Epithelial Cell Line ◽  
Huh7 Cells ◽  
Protein Expressions ◽  
Mesenchymal Transformation ◽  
E Cadherin

Abstract Background The aim was to investigate whether integrin α7 (ITGA7) influenced hepatocellular carcinoma (HCC) progression, and explore its effect on regulating endothelium-mesenchymal transformation (EMT).Methods ITGA7 mRNA and protein expressions in human normal liver epithelial cell line and HCC cell lines were determined by reverse transcription polymerase chain reaction (RT-qPCR) and western blot. ITGA7 siRNA (ITGA7-KD group) and nonsense siRNA (control group) were transfected into Huh7 cells and SUN449 cells. After transfection, ITGA7 mRNA and protein expressions (RT-qPCR and western blot), cell proliferation (Cell Counting Kit-8), apoptosis (Annexin V/Propidium Iodide assay), migration (Wound scratch assay) and invasion (Transwell assay) were determined. E-cadherin and α-SMA expressions (RT-qPCR and western blot) were determined.Results ITGA7 mRNA and protein expressions were increased in Li7, Huh7, SKHEP1 and SNU449 cells compared to THLE-3 cells. In both Huh7 and SNU449 cells, ITGA7 mRNA and protein expressions were decreased in ITGA7-KD group than control group after plasmids transfection, indicating the successful transfection. Then, cell proliferation was decreased at 48h and 72h; cell apoptosis rate was increased at 48h; cell migration rate was reduced at 24h; cell invasive count was decreased at 24h in ITGA7-KD group compared to control group. Furthermore, increased E-cadherin but decreased α-SMA mRNA and protein expressions were discovered in ITGA7-KD group than control group at 24h.Conclusions ITGA7 knockdown suppresses HCC progression and inhibits EMT process in HCC, indicating that ITGA7 might be a potential novel treatment target for HCC therapy.

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