Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways

Saroj K Shrestha,Se-Woong Kim,Yunjo Soh 대한수의학회 2023 Journal of Veterinary Science Vol.24 No.5

Background: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT’s mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. Objectives: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. Methods: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT’s effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. Results: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca++-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runt-related transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca2+-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. Conclusions: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis.

오가피(Eleutherococcus sessiliflorus )의 전연골성 ATDC5 세포의 분화 유도

스레스타사로즈쿠마 ( Saroj Kumar Shrestha ),송정빈 ( Jungbin Song ),이성현 ( Sung Hyun Lee ),이동헌 ( Donghun Lee ),김호철 ( Hocheol Kim ),소윤조 ( Yunjo Soh ) 대한본초학회 2022 大韓本草學會誌 Vol.37 No.1

Objectives : The process through which mesenchymal cells condense and differentiate into chondrocytes to form new bone is known as endochondral bone formation. Chondrogenic differentiation and hypertrophy are essential steps in bone formation and are influenced by various factors. The stem bark and root bark of Eleutherococcus sessiliflorus (ES) have been widely used to treat growth retardation and arthritis in traditional Korean Medicine. In this study, we aimed to investigate the possible role of the stem bark of ES in the stimulation of chondrogenic differentiation in clonal murine chondrogenic ATDC5 cells. Methods : In ATDC5 cells treated with ES extract, cell viability and extracellular matrix production were determined using CCK-8 assay and Alcian blue staining, respectively, and alkaline phosphatase activity was measured. We also examined mRNA and protein expression levels of genes related to chondrogenic expression in ATDC5 cells using reverse transcription-polymerase chain reaction and western blot analyses. Results : ES extract increased the accumulation of Alcian blue-stained cartilage nodules and alkaline phosphatase activity in ATDC5 cells. It increased the mRNA expressions of chondrogenic markers including bone sialoprotein (BSP), cartilage collagens, Runt-related transcription factor-2 (RUNX-2), osteocalcin (OCN), β-catenin, and bone morphogenetic protein-2 (BMP-2), as well as the protein expressions of β-catenin, RUNX-2, BMP-2, and alkaline phosphatase (ALP). Conclusion : Taken together, these results suggest that ES extract exhibits a chondromodulating activity and therefore may be a possible agent for the treatment of bone growth disorders.

Ubiquitin sepcific protease 2 acts as a key modulator for the regulation of cell cycle by adiponectin and leptin in cancer cells

( Saroj Nepal ),( Anup Shrestha ),( Pil Hoon Park ) 영남대학교 약품개발연구소 2016 영남대학교 약품개발연구소 연구업적집 Vol.26 No.-

Adiponectin and leptin, both produced from adipose tissue, cause cell cycle arrest and progression, re-spectively in cancer cells. Ubiquitin specific protease-2 (USP-2). a deubiquitinating enzyme, is known to impair proteasorne-induced degradation of cyelin D1, a critical cell cycle regulator. Herein, we investi-gated the effects of these adipokines on USP-2 expression and its potential role in the modulation of cell cycle. Treatment with globular adiponectin (gAcrp) decreased, whereas leptin increased USP-2 expres-sion both in human hepatoma and breast cancer cells. In addition, overexpression or gene silencing of USP-2 affected cyelin D1 expression and cell cycle progression/arrest by adipokines. Adiponectin and leptin also modulated in vitro proteasomal activity, which was partially dependent on USP-2 expression. Taken together. our results reveal that modulation of USP-2 expression plays a crucial role in cell cycle regu-lation by adipokines. Thus. USP-2 would be a promising therapeutic target for the modulation of cancer cell growth by adipokines.

Influence of the 3D modelling shapes from a 2D centrifugal pump model drawing on the pump performance and internal flow

Saroj Basnet(바스네드 사로즈),Ujjwal Shrestha(쉬레스트 우즈왈),Young-Do Choi(최영도) 한국유체기계학회 2022 한국유체기계학회 학술대회 논문집 Vol.2022 No.11

Screw centrifugal pump performances based on impeller inlet shape

Ujjwal Shrestha(쉬레스트 우즈왈),Saroj Basnet(바스네드사로즈),Young-Do Choi(최영도) 한국유체기계학회 2022 한국유체기계학회 학술대회 논문집 Vol.2022 No.6

Performance variation of a single channel pump by impeller design parameters

Ujjwal Shrestha(쉬레스트 우즈왈),Saroj Basnet(바스네드사로즈),Young-Do Choi(최영도) 한국유체기계학회 2022 한국유체기계학회 학술대회 논문집 Vol.2022 No.6

Critical Role of AMPK/Fox03A Axis in Globular Adiponectin-induced Cell Cycle Arrest and Apoptosis in Cancer Cells

( Anup Shrestha ),( Saroj Nepal ),( Mijin Kim ),( Jae Hoon Chang ),( Sang Hyun Kim ),( Gil Saeng Jeong ),( Chul Ho Jeong ),( Gyu Hwan Park ),( Sunghee Jung ),( Jaecheong Lim ),( Eunha Cho ),( Soyoung 영남대학교 약품개발연구소 2016 영남대학교 약품개발연구소 연구업적집 Vol.26 No.-

Adiponeectin predominantly secreted from adipose tissue has exhibited potent anti- proliferative properties in cancer cells via modulating cell cycle and apoptosis, FoxO3A. a Forkhead box 0 member of the transcription factor. plays a critical role in modulating expression of genes involved In cell de:uh andlor SUrviV31. In this study. we investigated the role of FoxO3A signaling in and-cancer activities of adiponectin. Herein. we have shown that treaunent with globular adlponectln (gAcrp) Increases p27 but decreases cyclinD I expression In human hepatoma (HepG2) and breasr (MCF-7) cancer cells. Gene ablation of Fox03A prevented gAcrp-induced increase In p27 and decreased in cyelin 0 I expression. and further ameliorated cell cycle arrest by gAcrp.lndicating 3 critical role of FoxO3A in gAcrp-induced cell cycle arrest of cancer cells. Moreover, treatment with gAcrp also induced caspase-Jf7 activation and increased Fas ligand (Fasl) expression ln both HepG2 and MCF-7 cells. Transfecucn with FoxO3A siRNA inhibited gAcrp-induced caspase-3/7 activadon and Fasl expression. suggening that FoxO3A sigltaling also plays an lmportant role In gAcrp.induced apcptosis of cancer cells, We also found that gene silencing of AMPK prevented gAcrp-induced nuclear translocation of Fox03A in HepG2 and MCF-7 cells. In addition. suppression of AMPK also blocked gAcrp-lnduced cell cyele arrest and further attenuated gAcrp-induced caspase-3/7 activation, indICating that AMPK signaling plays a pivotal role in both gAcrp-lnduced cell cycle arrest and apoprosis via acting as an upstream S1gnalirlg of Fox03A Taken together. our firldings demonstrated that AMPK/Fox03A axis plays a cardinal role in anu-protlferanve effect of adiponecun in cancer cells.

Effects of aloe-emodin on alveolar bone in Porphyromonas gingivalis-induced periodontitis rat model: a pilot study

Ming Yang,Saroj K Shrestha,소윤조,허석모 대한치주과학회 2022 Journal of Periodontal & Implant Science Vol.52 No.5

Purpose: Aloe-emodin (AE), a natural anthraquinone abundant in aloe plants and rhubarb (Rheum rhabarbarum), has long been used to treat chronic inflammatory diseases. However, AE’s underlying mechanisms in periodontal inflammation have not been fully elucidated. Acidic mammalian chitinase (AMCase) is a potential biomarker involved in bone remodeling. This study aimed to evaluate AE’s effect on periodontitis in rats and investigate AMCase expression. Methods: Eighteen Sprague-Dawley rats were separated into the following groups: healthy (group 1), disease (group 2), vehicle (group 3), AE high-dose (group 4), and AE low-dose (group 5). Porphyromonas gingivalis ligatures were placed in rats (groups 2–5) for 7 days. Groups 4 and 5 were then treated with AE for an additional 14 days. Saliva was collected from all groups, and probing pocket depth was measured in succession. Periodontal pocket tissues were subjected to histomorphometric analysis after the rats were sacrificed. Bone marrowderived macrophages and murine macrophages were stimulated with receptor activator of nuclear factor-κB ligand (RANKL) and treated with different concentrations of AE. AMCase expression was detected from the analysis of saliva, periodontal pocket tissues, and differentiated osteoclasts. Results: Among rats with P. gingivalis-induced periodontitis, the alveolar bone resorption levels and periodontal pocket depth were significantly reduced after treatment with AE. AMCase protein expression was significantly higher in the disease group than in the healthy control (P<0.05). However, AE inhibited periodontal inflammation by downregulating AMCase expression in saliva and periodontal pocket tissue. AE significantly reduced RANKLstimulated osteoclastogenesis by modulating AMCase (P<0.05). Conclusions: AE decreases alveolar bone loss and periodontal inflammation, suggesting that this natural anthraquinone has potential value as a novel therapeutic agent against periodontal disease.

Aloe-emodin inhibits osteogenic differentiation and calcification of mouse vascular smooth muscle cells

Sapkota, Mahesh,Shrestha, Saroj Kumar,Yang, Ming,Park, Young Ran,Soh, Yunjo Elsevier 2019 european journal of pharmacology Vol.865 No.-

<P><B>Abstract</B></P> <P>Vascular calcification increases the risk of morbidity and mortality in patients with cardiovascular diseases, chronic kidney diseases, and diabetes. However, viable therapeutic methods to target vascular calcification are limited. Aloe-emodin (AE), an anthraquinone is a natural compound found in the leaves of Aloe-vera. In this study, we investigated the underlying mechanism of AE in the calcification of vascular smooth muscle cells (VSMCs) and murine thoracic aorta. We demonstrate that AE repressed not only the phenotypes of Ca<SUP>2+</SUP> induced calcification but also level of calcium in VSMCs. AE has no effect on cell viability in VSMC cells. Alizarin red, von Kossa stainings and calcium quantification showed that Ca<SUP>2+</SUP> induced vascular calcification is significantly decreased by AE in a concentration-dependent manner. In contrast, AE attenuated Ca<SUP>2+</SUP> induced calcification through inhibiting osteoblast differentiation genes such as SMAD4, collagen 1α, osteopontin (OPN), Runt-related transcription factor (RUNX-2) and Osterix. AE also suppressed Ca<SUP>2+</SUP> induced osteoblast-related protein expression including collagen 1α, bone morphogenic protein 2 (BMP-2), RUNX-2 and smooth muscle actin (SMA). Furthermore, Alizarin red, von Kossa stainings and calcium quantification showed that AE significantly inhibited the calcification of <I>ex vivo</I> ring formation in murine thoracic aorta, and markedly inhibited vitamin D<SUB>3</SUB> induced medial aorta calcification <I>in vivo</I>. Taken together, our findings suggest that AE may have therapeutic potential for the prevention of vascular calcification program.</P>

Effects of fine particulate matter on bone marrow-conserved hematopoietic and mesenchymal stem cells: a systematic review

Bhattarai Govinda,Shrestha Saroj Kumar,Sim Hyun-Jaung,Lee Jeong-Chae,Kook Sung-Ho 생화학분자생물학회 2024 Experimental and molecular medicine Vol.56 No.-

The harmful effects of fine particulate matter ≤2.5 µm in size (PM2.5) on human health have received considerable attention. However, while the impact of PM2.5 on the respiratory and cardiovascular systems has been well studied, less is known about the effects on stem cells in the bone marrow (BM). With an emphasis on the invasive characteristics of PM2.5, this review examines the current knowledge of the health effects of PM2.5 exposure on BM-residing stem cells. Recent studies have shown that PM2.5 enters the circulation and then travels to distant organs, including the BM, to induce oxidative stress, systemic inflammation and epigenetic changes, resulting in the reduction of BM-residing stem cell survival and function. Understanding the broader health effects of air pollution thus requires an understanding of the invasive characteristics of PM2.5 and its direct influence on stem cells in the BM. As noted in this review, further studies are needed to elucidate the underlying processes by which PM2.5 disturbs the BM microenvironment and inhibits stem cell functionality. Strategies to prevent or ameliorate the negative effects of PM2.5 exposure on BM-residing stem cells and to maintain the regenerative capacity of those cells must also be investigated. By focusing on the complex relationship between PM2.5 and BM-resident stem cells, this review highlights the importance of specific measures directed at safeguarding human health in the face of rising air pollution.