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Alizarin red and Oil red O staining. Alizarin red staining of (A) control, (B) 10 -6 M ICA and (C) E2 treated cells. Oil red O staining of (D) control, (E) 10 -6 M ICA, and (F) adipocyte-induced cells (magnification, x200). ICA, icariin
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Icariin (ICA) is a major active ingredient in Herba epimedii, which is commonly used as a Chinese herbal medicine for the treatment of osteoporosis. Previous studies have revealed that ICA exerted a protective effect against bone loss and increased bone regeneration; however, the association between ICA and estrogen receptor (ER) signaling remains...
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... promotes osteogenic differentiation. The number of mineralized nodules can serve as measure of osteogenic differentiation. Compare to control, the number of mineralized nodules was increased in cells treated with ICA, and was also increased cells treated with E2 ( Fig. 3A-C). Fat droplets were identified by oil red O staining, and there was a reduction in the number and size of fat droplets in cells treated with ...
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... promotes osteogenic differentiation. The number of mineralized nodules can serve as measure of osteogenic differentiation. Compare to control, the number of mineralized nodules was increased in cells treated with ICA, and was also increased cells treated with E2 ( Fig. 3A-C). Fat droplets were identified by oil red O staining, and there was a reduction in the number and size of fat droplets in cells treated with ...
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With age, bone marrow mesenchymal stem cells (BMSC) have reduced ability of differentiating into osteoblasts but have increased ability of differentiating into adipocytes which leads to age-related bone loss. MicroRNAs (miRNAs) play major roles in regulating BMSC differentiation. This paper explored the role of miRNAs in regulating BMS...
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... It has been found to induce the expression of BMP-2 and BMP-4 mRNA in osteoblasts and significantly upregulated Osx at low doses [182]. Additionally, ICA promotes bone formation by stimulating the expression of pre-osteoblastic genes like Osx, RUNX2, and collagen type I [183]. ...
There is an increasing demand for innovative strategies that effectively promote osteogenesis and enhance bone regeneration. The critical process of bone regeneration involves the transformation of mesenchymal stromal cells into osteoblasts and the subsequent mineralization of the extracellular matrix, making up the complex mechanism of osteogenesis. Icariin’s diverse pharmacological properties, such as anti-inflammatory, anti-oxidant, and osteogenic effects, have attracted considerable attention in biomedical research. Icariin, known for its ability to stimulate bone formation, has been found to encourage the transformation of mesenchymal stromal cells into osteoblasts and improve the subsequent process of mineralization. Several studies have demonstrated the osteogenic effects of icariin, which can be attributed to its hormone-like function. It has been found to induce the expression of BMP-2 and BMP-4 mRNAs in osteoblasts and significantly upregulate Osx at low doses. Additionally, icariin promotes bone formation by stimulating the expression of pre-osteoblastic genes like Osx, RUNX2, and collagen type I. However, icariin needs to be effectively delivered to bone to perform such promising functions.
Encapsulating icariin within nanoplatforms holds significant promise for promoting osteogenesis and bone regeneration through a range of intricate biological effects. When encapsulated in nanofibers or nanoparticles, icariin exerts its effects directly at the cellular level. Recalling that inflammation is a critical factor influencing bone regeneration, icariin's anti-inflammatory effects can be harnessed and amplified when encapsulated in nanoplatforms. Also, while cell adhesion and cell migration are pivotal stages of tissue regeneration, icariin-loaded nanoplatforms contribute to these processes by providing a supportive matrix for cellular attachment and movement. This review comprehensively discusses icariin-loaded nanoplatforms used for bone regeneration and osteogenesis, further presenting where the field needs to go before icariin can be used clinically.
... Interestingly, an inhibitory effect (p < 0.05) was noted for 10 − 8 mol/L − 10 − 5 mol/L on the fourth and sixth days. It is noteworthy that some studies reported no inhibitory effects of ICA on rBMSCs17 , which could be in uenced by experimental conditions, disparate action durations, and distinct experimental methodologies. In the ow apoptosis assay, both ICA and ICSII exhibited inhibitory effects on apoptosis across all concentrations when compared with the control group. ...
ICA (icariin) serves as the primary biologically active compound in traditional Chinese medicine Epimedium, while Icariside II (ICSII) represents one of its gastrointestinal metabolites.Although ICA and ICSII have demonstrated osteogenic differentiation- promoting effects on BMSCs, there is limited literature comparing their effects and underlying mechanisms. This study aimed to compare the osteogenic effects of Icariin and Icarisin II, along with their respective osteogenic mechanisms. In this study, we initially determined the optimal concentrations of Icariin (10 − 5 mol/L) and Icariin II (10 − 6 mol/L) for inducing BMSC osteogenic differentiation using CCK8, ALP activity assay, and flow apoptosis assay. Subsequently, we compared the vascularization and osteogenic capacity of the two groups through alizarin red staining assay, ELISA assay, Western Blot, and RY-PCR. Subsequently, we assessed the phosphorylated and non-phosphorylated expression of JNK, ERK1/2, p38, and AKT at different time intervals. We observed their phosphorylated expression and the expression of angiogenic/osteogenic markers after blocking with their corresponding inhibitors. It was observed that both the Icariin and Icariin II groups promoted the expression of osteogenic/angiogenic markers Runx-2, OCN, OPN, VEGF, and Ang1. While there was no significant difference in their osteogenic abilities, ICSII exhibited a stronger promotion of angiogenic differentiation markers, Ang1 and VEGF, compared to ICA. Additionally, it was observed that both ICA and ICSII could activate ERK1/2 phosphorylation, thereby further promoting the osteogenic/angiogenic differentiation of rBMSCs through the activation of the MAPK/ERK1/2 signaling pathway.
... ICA activated the ER signaling pathway to promote osteoblast differentiation and suppressed adipogenic differentiation of MSCs by greatly upregulating the expression of both ERα and ERβ. 89 Exercise enhanced this process via upregulating the expression of ERα and then strengthening the Akt/β-catenin pathway. 66 Specific MAPK inhibitors, including the JNK inhibitor SP600125 and the ERK inhibitor U0126, reduced the effects of ICA on osteoblast differentiation. ...
Osteoporosis is a metabolic skeletal disorder characterized by increased fragility and fracture risk as s result of reduced bone mineral density and microstructural destruction and caused a heavy burden on families and society. Current medicines, on the other hand, have some limitations, with side effects and doubts regarding long-term efficacy being highlighted. Studies seeking for natural constituents as potential treatment options therefore come into focus. Icariin is a phytochemical derived from a traditional Chinese medicine, Herba epimedium, that has been used to treat orthopedic disorders in ancient China for thousands of years, including osteoporosis, osteoarthritis, and fracture. Icariin belongs to a category of prenylated flavonoids and has been shown to help reduce osteoporosis bone loss while having relatively low side effects. Icariin's anti-osteoporosis properties manifest in a variety of ways, like promoting osteogenesis, suppressing osteoclastogenesis and bone resorption, regulating migration, proliferation, and differentiation of mesenchymal stem cells, enhancing angiogenesis, anti-inflammation, and antioxidation. These procedures entail a slew of critical signaling pathways, such as PPARγ, ERα/AKT/β-catenin, and MAPK. Therefore, icariin can be an applicable alternative to improve osteoporosis although the underlying mechanisms have yet to be fully understood. In this study, we searched using the terms “icariin” and “osteoporosis,” and included 64 articles meeting the inclusion criteria and reviewed the research of icariin in anti-osteoporosis over the last 10 years, and discussed new prospects for future study. Therefore, this review may provide some references for further studies.
... A recent study found marked activation of ERα and elevated osteogenic activity in both pre-osteoblastic cells and mature osteoblasts after ICA treatment (Zhou et al., 2021), indicating potential links between ERα and the bone preserving effect of ICA. Previous studies also reported that ICA stimulated osteoblast differentiation of bone marrow stromal cells via activation of the ERα signaling pathway, which could be reversed by ERα antagonist (Wei et al., 2016;Li et al., 2018). Additionally, bioinformatic analysis in the present study found that ERα has a close relationship with TNF-α and IL-6, indicating that ERα is a potential regulatory target for CoCrMo particle-induced macrophage activation. ...
Periprosthetic osteolysis is one of the major long-term complications following total joint replacement. Its cause is widely accepted to be wear particle-induced activation of inflammatory macrophages. No effective strategy for the prevention and treatment of periprosthetic osteolysis is yet available. Recently, considerable evidence has shown that icariin effectively protects against estrogen deficiency-related bone loss and bone deterioration. However, the molecular mechanism underlying the inhibitory effect of icariin on wear particle-induced periprosthetic osteolysis is not yet clear. In this study, nanoscale CoCrMo wear particles were obtained by high-vacuum three-electrode direct current from the femoral head implant of a patient diagnosed with aseptic loosening. The effects of icariin on wear particle-induced expression of proinflammatory factors, NF-κB signaling modulation, osteolysis, and estrogen receptor α (ERα) activation were evaluated in vitro and in vivo using bone marrow-derived macrophages and C57/BL6J mice, respectively. A possible link between ERα and the protective effect of icariin was further studied using an ERα antagonist and the ERα-siRNA interference. Chemical composition analysis showed that Cr and Co were the major metallic elements of the nanoscale particles, with a mean size of 150.2 ± 37.4 nm for the CoCrMo particles. Following icariin treatment, significant decreases were observed in CoCrMo wear particle-induced TNF-α and IL-6 mRNA expression in BMDMs, and osteolysis in mice calvaria. Marked decreases in the protein expression level of p-IKKβ, p-p65 and p-IκBα were also observed, together with significant decreases in the nuclear import of P65 and macrophage M1 polarization. RNA sequencing revealed that ERα was closely associated with TNF-α and IL-6 in wear particle-stimulated macrophages. Furthermore, marked increases in phospho-ERα Ser118 and phospho-ERα Ser167 protein expression and the nuclear import of ERα were also found in the icariin group. The protective effects of icariin on CoCrMo particle-induced mouse calvarial osteolysis and on the inflammation response in BMDMs were reversed by ERα antagonist and by ERα-siRNA interference. In conclusion, icariin attenuates wear particle-induced inflammation and osteolysis via down-regulation of the ERα-mediated NF-κB signaling pathway in macrophages. The potential application of icariin as a non-hormonal therapy for wear particle-induced periprosthetic osteolysis is worthy of further investigation.
... 154 Icariin activates the ESR1 signaling pathway; our network pharmacology analysis revealed that its antidepressant effect may be related to the regulation of estrogen signaling. 155,157 On the other hand, the decreased GSK-3β phosphorylation detected in the platelets of patients treated for depression implies that GSK-3β hyperactivation contributes to the pathophysiology of depressive disorder; this is underscored by the observation that blocking GSK-3β activity enhanced the effects of antidepressant therapy. 75,158 Icariin was been reported to inhibit GSK-3β in part by activating the PI3K/AKT pathway, 76 which may be the mechanism by which it alleviates depressive symptoms. ...
Icariin is a biologically active substance in Epimedii herba that is used for the treatment of neurologic disorders. However, a comprehensive analysis of the molecular mechanisms of icariin is lacking. In this review, we present a brief history of the use of icariin for medicinal purposes; describe the active chemical components of Epimedii herba; and examine the evidence from experimental studies that have uncovered molecular targets of icariin in different diseases. We also constructed a protein–protein interaction network and carried out Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional enrichment analyses to predict the therapeutic actions of icariin in nervous system diseases including Alzheimer disease, Parkinson disease, ischemic stroke, depressive disorder, multiple sclerosis, glioblastoma, and hereditary spastic paraplegias. The results of our analyses can guide future studies on the application of icariin to the treatment of neurologic disorders.
... 154 Icariin activates the ESR1 signaling pathway; our network pharmacology analysis revealed that its antidepressant effect may be related to the regulation of estrogen signaling. 155,157 On the other hand, the decreased GSK-3β phosphorylation detected in the platelets of patients treated for depression implies that GSK-3β hyperactivation contributes to the pathophysiology of depressive disorder; this is underscored by the observation that blocking GSK-3β activity enhanced the effects of antidepressant therapy. 75,158 Icariin was been reported to inhibit GSK-3β in part by activating the PI3K/AKT pathway, 76 which may be the mechanism by which it alleviates depressive symptoms. ...
Shuangqiu Wang,1– 3,* Jiarui Ma,4,* Yanqi Zeng,5,* Guowei Zhou,6 Yuxuan Wang,1– 3 Wenjuan Zhou,5 Xiaohe Sun,5 Minghua Wu1,5 1Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, Jiangsu, People’s Republic of China; 2Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing, 210046, People’s Republic of China; 3State Key Laboratory Cultivation Base for TCM Quality and Efficacy, Nanjing University of Chinese Medicine, Nanjing, 210046, Jiangsu, People’s Republic of China; 4Provincial Key Laboratory of Drug Target and Drug for Degenerative Disease, School of Medicine and Holistic Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing, 210046, Jiangsu, People’s Republic of China; 5First Clinical Medical School, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210046, Jiangsu, People’s Republic of China; 6Department of General Surgery, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, Jiangsu, People’s Republic of China*These authors contributed equally to this workCorrespondence: Minghua WuDepartment of Neurology, Jiangsu Provincial Hospital of Chinese Medicine, The Affiliated Hospital of Nanjing University of Chinese Medicine, No. 155 Hanzhong Road, Qinhuai District, Nanjing, Jiangsu, People’s Republic of ChinaTel +86-13951786719Fax +86 25 8661 7141Email mhuawu@163.comAbstract: Icariin is a biologically active substance in Epimedii herba that is used for the treatment of neurologic disorders. However, a comprehensive analysis of the molecular mechanisms of icariin is lacking. In this review, we present a brief history of the use of icariin for medicinal purposes; describe the active chemical components of Epimedii herba; and examine the evidence from experimental studies that have uncovered molecular targets of icariin in different diseases. We also constructed a protein–protein interaction network and carried out Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional enrichment analyses to predict the therapeutic actions of icariin in nervous system diseases including Alzheimer disease, Parkinson disease, ischemic stroke, depressive disorder, multiple sclerosis, glioblastoma, and hereditary spastic paraplegias. The results of our analyses can guide future studies on the application of icariin to the treatment of neurologic disorders.Keywords: Epimedii herba, icariin, network pharmacology, literature review, nervous system diseases
... • Prevents adipogenic differentiation and induces of osteoblast differentiation through the stimulation of the ER signaling pathway. 113 ...
Medicinal plants have always been utilized for the prevention and treatment of the spread of different diseases all around the world. To name some traditional medicine that has been used over centuries, we can refer to phytochemicals such as naringin, icariin, genistein, and resveratrol gained from plants. Osteogenic differentiation and mineralization of stem cells can be the result of specific bioactive compounds from plants. One of the most appealing choices for therapy can be mesenchymal stem cells (MSCs) because it has a great capability of self‐renewal and differentiation into three descendants, namely, endoderm, mesoderm, and ectoderm. Stem cell gives us the glad tidings of great advances in tissue regeneration and transplantation field for treatment of diseases. Using plant bioactive phytochemicals also holds tremendous promises in treating diseases such as osteoporosis. The purpose of the present review article thus is to investigate what are the roles and consequences of phytochemicals on osteogenic differentiation of MSCs.
... In this study, we constructed an ERα-deficient rBMSC model. The results of protein, mRNA and alizarin red S staining showed that ERα deficiency suppressed the osteogenic ability of rBMSCs, which was in accord with the results of our previous study, which showed that the osteogenic and adipogenic differentiation ability of rBMSCs could be regulated by ER signaling (Li et al., 2018). ...
Circular RNA (circRNA) participates in regulation of gene transcription, while estrogen receptor alpha (ERα) and quercetin (QUE) positively regulate bone formation, but little is known about the correlation among circRNA, ERα and QUE. In this experiment, we created an ERα-deficient rBMSC model treated with QUE and evaluated the effects of ERα or QUE on rBMSCs, then analyzed differentially-expressed circRNAs by RNA-Seq and bioinformatics. The results showed that ERα deficiency constrained osteogenic differentiation and stimulated adipocytic differentiation of rBMSCs, while QUE abrogated those effects. We identified 136 differentially-expressed circRNAs in the Lv-shERα group and 120 differentially-expressed circRNAs in the Lv-shERα + QUE group. Thirty-two circRNAs retroregulated by ERα and QUE were involved in Rap1 and Wnt signaling, and four of them together sponged miR-326-5p, the target genes of which are osteogenic and adipogenic differentiation factors. Further study showed that over-expressed miR-326-5p could stimulate osteogenic differentiation, while attenuating adipogenic differentiation of rBMSCs. Therefore, we concluded that ERα and QUE might regulate the differentiation of rBMSCs through the circRNA–miR-326-5p–mRNA axis.
... Epimedium brevicornu Maxim., significantly promoted osteogenic differentiation by increasing ALP activity and Runx2, β-catenin, type I collagen (COLI), osteocalcin (OCN), and osteopontin (OPN) expression in rat BMSCs [39,40]. Moreover, icariin had a bidirectional regulation effect on promoting the differentiation of bone marrow mesenchymal stem cells or bone marrow stromal cells into osteoblasts and inhibiting the differentiation into adipocytes [41,42]. Micro-CT analysis showed that icariin relieved the loss of cancellous bone of the distal femur in OVX mice [41]. ...
Stem cells (SCs) are special types of cells with the ability of self-renewal and multidirectional differentiation. As the organism ages, the ability to maintain homeostasis and regeneration deteriorates and the number and activity of stem cells decline. Theoretically, the restoration of stem cells might reverse aging. However, due to their own aging, donor-derived immune rejection, and difficulties in stem cell differentiation control, a series of problems need to be solved to realize the potential for clinical application of stem cells. Chinese herbal medicine is a nature drug library which is suitable for the long-term treatment of aging-related diseases. Modern pharmacological studies have revealed that many active ingredients of Chinese herbal medicines with the effect of promoting stem cells growth and differentiation mainly belong to “reinforcing herbs.” In recent years, exploration of natural active ingredients from Chinese herbal medicines for delaying aging, improving the stem cell microenvironment, and promoting the proliferation and differentiation of endogenous stem cells has attracted substantial attention. This article will focus on active ingredients from Chinese herbs-mediated differentiation of stem cells into particular cell type, like neural cells, endothelial cells, cardiomyocytes, and osteoblasts. We will also discuss the effects of these small molecules on Wnt, Sonic Hedgehog, Notch, eNOS-cGMP, and MAP kinase signal transduction pathways, as well as reveal the role of estrogen receptor α and PPAR γ on selectively promoting or inhibiting stem cells differentiation. This review will provide new insights into the health aging strategies of active ingredients in Chinese herbal medicine in regenerative medicine.
... A recent study revealed MIAT was an estrogen-inducible lncRNA and its expression was positively related to estrogen receptor (Li et al., 2018b). There was accumulating evidence to reveal that exposure of bone marrow stem cells to icariin or flavonoids of Herba Epimedii inhibited adipogenic differentiation, exhibiting decreased adipocyte numbers and downregulated mRNA expression of adipogenic differentiation markers, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein α (C/EBPα) (Li et al., 2018c;Zhang et al., 2015); while treatment of bone marrow stem cells with estrogen receptor antagonist ICI182780 revered the effects of Herba Epimedii ingredient and promoted adipogenesis (Li et al., 2018c;Zhang et al., 2015). The study of Ihunnah et al. (2014) also demonstrated activation of estrogen receptor in ASCs inhibited adipogenesis by decreasing the recruitment of the adipogenic PPARγ onto its target gene promoters, whereas the use of estrogen receptor antagonism ICI 182780 or knockdown of estrogen receptor-α via lentiviral shRNA enhanced adipogenesis by increasing the expression of PPARγ. ...
... A recent study revealed MIAT was an estrogen-inducible lncRNA and its expression was positively related to estrogen receptor (Li et al., 2018b). There was accumulating evidence to reveal that exposure of bone marrow stem cells to icariin or flavonoids of Herba Epimedii inhibited adipogenic differentiation, exhibiting decreased adipocyte numbers and downregulated mRNA expression of adipogenic differentiation markers, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein α (C/EBPα) (Li et al., 2018c;Zhang et al., 2015); while treatment of bone marrow stem cells with estrogen receptor antagonist ICI182780 revered the effects of Herba Epimedii ingredient and promoted adipogenesis (Li et al., 2018c;Zhang et al., 2015). The study of Ihunnah et al. (2014) also demonstrated activation of estrogen receptor in ASCs inhibited adipogenesis by decreasing the recruitment of the adipogenic PPARγ onto its target gene promoters, whereas the use of estrogen receptor antagonism ICI 182780 or knockdown of estrogen receptor-α via lentiviral shRNA enhanced adipogenesis by increasing the expression of PPARγ. ...
Background
Injection of adipose-derived stem cells (ASCs) is a promising treatment for facial contour deformities. However, its treatment mechanisms remain largely unknown. The study aimed to explain the molecular mechanisms of adipogenic differentiation from ASCs based on the roles of long noncoding RNAs (lncRNAs).
Methods
Datasets of mRNA–lncRNA ( GSE113253 ) and miRNA ( GSE72429 ) expression profiling were collected from Gene Expression Omnibus database. The differentially expressed genes (DEGs), lncRNAs (DELs) and miRNAs (DEMs) between undifferentiated and adipocyte differentiated human ASCs were identified using the Linear Models for Microarray Data method. DELs related co-expression and competing endogenous RNA (ceRNA) networks were constructed. Protein–protein interaction (PPI) analysis was performed to screen crucial target genes.
Results
A total of 748 DEGs, 17 DELs and 51 DEMs were identified. A total of 13 DELs and 279 DEGs with Pearson correlation coefficients > 0.9 and p -value < 0.01 were selected to construct the co-expression network. A total of 151 interaction pairs among 112 nodes (10 DEMs; eight DELs; 94 DEGs) were obtained to construct the ceRNA network. By comparing the lncRNAs and mRNAs in two networks, five lncRNAs (SNHG9, LINC02202, UBAC2-AS1, PTCSC3 and myocardial infarction associated transcript (MIAT)) and 32 genes (i.e., such as phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), protein tyrosine phosphatase receptor type B (PTPRB)) were found to be shared. PPI analysis demonstrated PIK3R1 , forkhead box O1 (FOXO1; a transcription factor) and estrogen receptor 1 (ESR1) were hub genes, which could be regulated by the miRNAs that interacted with the above five lncRNAs, such as LINC02202-miR-136-5p-PIK3R1, LINC02202-miR-381-3p-FOXO1 and MIAT-miR-18a-5p-ESR1. LINC02202 also could directly co-express with PIK3R1. Furthermore, PTPRB was predicted to be modulated by co-expression with LINC01119.
Conclusion
MIAT, LINC02202 and LINC01119 may be potentially important, new lncRNAs associated with adipogenic differentiation of ASCs. They may be involved in adipogenesis by acting as a ceRNA or co-expressing with their targets.
