Irisin Treatment Research Articles

Abstract TP231: Irisin, Elicited By Low Frequency Whole Body Vibration Or Exogenously, Improves Post-Stroke Cognition And Reduces Infarct Volume In Middle-Aged Rats

Significance: Low frequency whole body vibration (LFV) at 40 Hz, a low impact form of exercise, for a month following mild transient middle-cerebral artery occlusion (tMCAO) reduces infarct volume and improves motor function in reproductively senescent, middle-aged female rats (1). In humans, LFV was shown to increase circulating levels of irisin, a skeletal muscle-derived hormone. Irisin has also been shown to play a crucial role in preserving mitochondrial function, preventing oxidative stress, and elevating expression of BDNF, among other neuroprotective measures. The current study aims to investigate the efficacy of LFV in ameliorating post-tMCAO cognitive deficits and to determine the putative role of irisin in conferring the benefits of LFV in middle-aged rats. Methods: Middle-aged rats of both sexes (5-8) were randomly assigned to tMCAO (90 min), or sham surgery followed by exposure to either LFV (twice a day for 15 min each for 5 days a week over a month) or no LFV treatment groups. Following the last LFV treatment, rats were tested for hippocampus-dependent learning and memory using a water maze followed harvesting brain and blood samples for histopathological and inflammatory marker analyses, respectively. In a parallel experiment in the absence of LFV, middle-aged female rats were randomly assigned to either saline or irisin treatment following tMCAO. Recombinant irisin was purchased from PeproTech (Rocky Hill, NJ). Rats were treated with irisin (0.2 μg/g BW; IP) or saline for a month followed by their brains were assessed by histopathology. Results: Post-tMCAO LFV significantly lessens cognitive deficits in rats of both sexes. It also significantly decreased circulating pro-inflammatory cytokines and increased serum levels of native irisin. Quantification of infarct volume irisin-treated rats demonstrated that, compared to saline, infarct volume was significantly reduced. Saline treatment resulted in 234 ± 30 mm 3 , while irisin treatment yielded 128 ± 34 mm 3 (p<0.05) of infarct volume. Conclusion: Irisin, either as elicited by LFV or administered exogenously without LFV, regulates brain metabolism and inflammation and its beneficial effects can be stimulated by LFV.Reference: 1. Raval, A.P., et al., Int J Mol Sci, 2018. 19 (9).

Renal failure suppresses muscle irisin expression, and irisin blunts cortical bone loss in mice.

BackgroundChronic renal failure induces bone mineral disorders and sarcopenia. Skeletal muscle affects other tissues, including bone, by releasing myokines. However, the effects of chronic renal failure on the interactions between muscle and bone remain unclear.MethodsWe investigated the effects of renal failure on bone, muscle, and myokines linking muscle to bone using a mouse 5/6 nephrectomy (Nx) model. Muscle mass and bone mineral density (BMD) were analysed by quantitative computed tomography 8 weeks after Nx.ResultsNephrectomy significantly reduced muscle mass in the whole body (12.1% reduction, P < 0.05), grip strength (10.1% reduction, P < 0.05), and cortical BMD at the femurs of mice (9.5% reduction, P < 0.01) 8 weeks after surgery, but did not affect trabecular BMD at the femurs. Among the myokines linking muscle to bone, Nx reduced the expression of irisin, a proteolytic product of fibronectin type III domain‐containing 5 (Fndc5), in the gastrocnemius muscles of mice (38% reduction, P < 0.01). Nx increased myostatin mRNA levels in the gastrocnemius muscles of mice (54% increase, P < 0.01). In simple regression analyses, cortical BMD, but not trabecular BMD, at the femurs was positively related to Fndc5 mRNA levels in the gastrocnemius muscles of mice (r = 0.651, P < 0.05). The weekly administration of recombinant irisin to mice ameliorated the decrease in cortical BMD, but not muscle mass or grip strength, induced by Nx (6.2% reduction in mice with Nx vs. 3.3% reduction in mice with Nx and irisin treatment, P < 0.05).ConclusionsThe present results demonstrated that renal failure decreases the expression of irisin in the gastrocnemius muscles of mice. Irisin may contribute to cortical bone loss induced by renal failure in mice as a myokine linking muscle to bone.

Integrin αV Mediates the Effects of Irisin on Human Mature Adipocytes

Introdution and Aims: The myokine irisin is critical to modulating adipocytes thermogenesis and influence whole-body metabolism. However, whether there is difference in the effects of irisin on adipocytes derived from different depots remains unknown, and the receptor of irisin on adipocytes is still unclear. In this study, we determine the browning effect of irisin on adipocytes of subcutaneous and visceral human adipose tissue and explore the possibility that integrin αV was the receptor of irisin on human adipocytes. Methods: Human adipose-derived stem cells were isolated from human subcutaneous and visceral white adipose tissues and induced to differentiate into mature adipocytes, and the expression of UCP1 and thermogenic genes in mature adipocytes were examined with or without irisin treatment and compared between groups of different adiposity and different spots. Immunoprecipitation analysis was used to detect the interaction between irisin and integrin αV on adipocytes, and the protein expression of integrin αV in adipocytes was also compared between groups of different adiposity and anatomic position. Results: Irisin treatment could increase the expression level of beige adipocyte marker protein UCP1 and specific thermogenic genes in mature adipocytes derived from subcutaneous white adipose tissue but not in visceral adipose tissue. The results of immunoprecipitation showed that irisin could be attached to integrin αV on mature adipocytes, and there was no significant difference in the gene and protein expression of integrin αV in adipocytes, either derived from subcutaneous and visceral adipose tissue, or derived from obese and normal-weight individuals. Conclusion: The results of the present study indicated that irisin contributed to the transformation of mature white adipocytes to beige adipocytes in human subcutaneous adipose tissue but not in visceral adipose tissue. Integrin αV may mediate the browning effects of irisin on human mature adipocytes, which could provide the potential therapeutic targets for obesity and metabolic syndrome by promoting human brown adipose tissue activity.

Irisin injection mimics exercise effects on the brain proteome.

Physical inactivity can endanger human health and increase the incidence of neurodegenerative disease. Exercise has tremendous beneficial effects on brain health and cognitive function, especially in older adults. It also improves brain-related outcomes in depression, epilepsy and neurodegenerative disorders, such as Parkinson's disease and Alzheimer's disease. Irisin is a mediator of the beneficial effects of exercise. This study aimed to assess the proteome alterations in adult male National Maritime Research Institute (NMRI) mice brain tissue upon three different conditions including endurance exercise, resistance exercise and irisin injection. Quantification of irisin levels in blood was performed using irisin-ELISA Kit. Quantification and identification of proteins via two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS)/MS showed the alteration of at least 21 proteins due to different treatments. Cellular pathway analysis revealed common beneficial effects of sole irisin treatment and different exercise procedures suggesting the capability of irisin injection to substitute the exercise when physical activity is not possible.

Irisin Protects the Human Placenta from Oxidative Stress and Apoptosis via Activation of the Akt Signaling Pathway.

Irisin is a newly discovered exercise-mediated polypeptide hormone. Irisin levels increase during pregnancy however, women with preeclampsia (PE) have significantly lower levels of Irisin compared to women of healthy pregnancies. Even though many studies suggest a role of Irisin in pregnancy, its function in the human placenta is unclear. In the current study, we aimed to understand key roles of Irisin through its ability to protect against apoptosis is the preeclamptic placenta and in ex vivo and in vitro models of hypoxia/re-oxygenation (H/R) injury. Our studies show that Irisin prevents cell death by reducing pro-apoptotic signaling cascades, reducing cleavage of PARP to induce DNA repair pathways and reducing activity of Caspase 3. Irisin caused an increase in the levels of anti-apoptotic BCL2 to pro-apoptotic BAX and reduced ROS levels in an in vitro model of placental ischemia. Furthermore, we show that Irisin treatment acts through the Akt signaling pathway to prevent apoptosis and enhance cell survival. Our findings provide a novel understanding for the anti-apoptotic and pro-survival properties of Irisin in the human placenta under pathological conditions. This work yields new insights into placental development and disease and points towards intervention strategies for placental insufficiencies, such as PE, by protecting and maintaining placental function through inhibiting hypoxic ischemia-induced apoptosis.

Irisin Stimulates the Release of CXCL1 From Differentiating Human Subcutaneous and Deep-Neck Derived Adipocytes via Upregulation of NFκB Pathway.

Thermogenic brown and beige adipocytes might open up new strategies in combating obesity. Recent studies in rodents and humans have indicated that these adipocytes release cytokines, termed “batokines”. Irisin was discovered as a polypeptide regulator of beige adipocytes released by myocytes, primarily during exercise. We performed global RNA sequencing on adipocytes derived from human subcutaneous and deep-neck precursors, which were differentiated in the presence or absence of irisin. Irisin did not exert an effect on the expression of characteristic thermogenic genes, while upregulated genes belonging to various cytokine signaling pathways. Out of the several upregulated cytokines, CXCL1, the highest upregulated, was released throughout the entire differentiation period, and predominantly by differentiated adipocytes. Deep-neck area tissue biopsies also showed a significant release of CXCL1 during 24 h irisin treatment. Gene expression data indicated upregulation of the NFκB pathway upon irisin treatment, which was validated by an increase of p50 and decrease of IκBα protein level, respectively. Continuous blocking of the NFκB pathway, using a cell permeable inhibitor of NFκB nuclear translocation, significantly reduced CXCL1 release. The released CXCL1 exerted a positive effect on the adhesion of endothelial cells. Together, our findings demonstrate that irisin stimulates the release of a novel adipokine, CXCL1, via upregulation of NFκB pathway in neck area derived adipocytes, which might play an important role in improving tissue vascularization.

Deletion of PRAK Mitigates the Mitochondria Function and Suppresses Insulin Signaling in C2C12 Myoblasts Exposed to High Glucose.

Background: p38 regulated/activated protein kinase (PRAK) plays a crucial role in modulating cell death and survival. However, the role of PRAK in the regulation of metabolic stress remains unclear. We examined the effects of PRAK on cell survival and mitochondrial function in C2C12 myoblasts in response to high glucose stresses. Methods: PRAK of C2C12 myoblasts was knocked out by using CRISPR/Cas-9 genome editing technology. Both wild type and PRAK−/− C2C12 cells were exposed to high glucose at the concentration of 30 mmol/L to induce metabolic stress. The effect of irisin, an adipomyokine, on both wild type and PRAK−/− cells was determined to explore its relationship with RPAK. Cell viability, ATP product, glucose uptake, mitochondrial damage, and insulin signaling were assessed. Results: PRAK knockout decreased C2C12 viability in response to high glucose stress as evident by MTT assay in association with the reduction of ATP and glucose uptake. PRAK knockout enhanced apoptosis of C2C12 myoblasts in response to high glucose, consistent with an impairment in mitochondrial function, by decreasing mitochondrial membrane potential. PRAK knockout induced impairment of mitochondrial and cell damage were rescued by irisin. PRAK knockout caused decrease in phosphorylated PI3 kinase at Tyr 485, IRS-1 and AMPKα and but did not affect non-phosphorylated PI3 kinase, IRS-1 and AMPKα signaling. High glucose caused the further reduction of phosphorylated PI3 kinase, IRS-1 and AMPKα. Irisin treatment preserved phosphorylated PI3 kinase, IRS-1by rescuing PRAK in high glucose treatment. Conclusion: Our finding indicates a pivotal role of PRAK in preserving cellular survival, mitochondrial function, and high glucose stress.

Irisin alleviates obesity-related spermatogenesis dysfunction via the regulation of the AMPK\u03b1 signalling pathway

BackgroundInfertility is a common complication in obese men. Oxidative stress and testicular apoptosis play critical roles in obesity-induced spermatogenesis dysfunction. It has been reported that irisin, an exercise-induced myokine, may attenuate oxidative damage and testicular apoptosis in several diseases; however, its role in obesity-induced spermatogenesis dysfunction remains unclear. The purpose of this study was to investigate the role and underlying mechanism of irisin in obesity-induced dysfunction of spermatogenesis.MethodsMale mice were fed a high-fat diet (HFD) for 24 weeks to establish a model of obesity-induced spermatogenesis dysfunction. To explore the effects of irisin, mice were subcutaneously infused with recombinant irisin for 8 weeks beginning at 16 weeks after starting a HFD. To confirm the role of AMP-activated protein kinase α (AMPKα), AMPKα-deficient mice were used.ResultsThe data showed decreased serum irisin levels in obese patients, which was negatively correlated with sperm count and progressive motility. Irisin was downregulated in the plasma and testes of obese mice. Supplementation with irisin protected against HFD-induced spermatogenesis dysfunction and increased testosterone levels in mice. HFD-induced oxidative stress, endoplasmic reticulum (ER) stress and testicular apoptosis were largely attenuated by irisin treatment. Mechanistically, we identified that irisin activated the AMPKα signalling pathway. With AMPKα depletion, we found that the protective effects of irisin on spermatogenesis dysfunction were abolished in vivo and in vitro.ConclusionsIn conclusion, we found that irisin alleviated obesity-related spermatogenesis dysfunction via activation of the AMPKα signalling pathway. Based on these findings, we hypothesized that irisin is a potential therapeutic agent against obesity-related spermatogenesis dysfunction.

Irisin Attenuates Osteogenic Differentiation of Mouse Mesenchymal Stem Cells via Suppressing Bone Morphogenetic Protein-2 Signaling

This study assessed the impact of irisin on mouse mesenchymal stem cells (MSCs) differentiation and the underlying mechanisms. Bone marrow mesenchymal stem cells (BMMSCs) and mesenchymal progenitor cells (KUSA-A1 cells) were isolated from mice and inoculated into petri dishes. Cell proliferation and apoptosis under different concentrations of irisin were detected by MTT. Irisin (1 μM) with nontoxic dose concentration was selected for subsequent experiments. Cells were exposed to 1 μM irisin, osteogenic differentiation was detected by von kossa stain, we employed oil red o stain to test adipocyte differentiation, Alcian blue stain to determine chondrocyte differentiation. BMP-2 expression was analyzed by immunocytochemical staining. Finally, signal transduction pathways and expression and transcription levels of osteogenic differentiation markers in irisin-treated cells were detected by protein imprinting and PCR. BMMSCs and KUSA-A1 cells displayed significantly suppressed osteogenic differentiation and reduced formation of extracellular mineralized matrix, while BMMSCs presented unaffected adipocyte differentiation and chondrocyte differentiation. 1 μM Irisin did not exert cytotoxicity. Further, irisin treatment abated osteogenic differentiation makers Runx2, Osterix, Osteocalcin, Osteopontin, alkaline phosphatase expression. Finally, BMP-2/Smads signaling related molecules (BMP-2, Smad1, Smad4, Smad5 and Smad8) levels were reduced after Irisin treatment. Irisin might inhibit osteoblast differentiation by modulating BMP-2/Smads axis.

Irisin ameliorates doxorubicin-induced cardiac perivascular fibrosis through inhibiting endothelial-to-mesenchymal transition by regulating ROS accumulation and autophagy disorder in endothelial cells

The dose-dependent toxicity to cardiomyocytes has been well recognized as a central characteristic of doxorubicin (DOX)-induced cardiotoxicity (DIC), however, the pathogenesis of DIC in the cardiac microenvironment remains elusive. Irisin is a new hormone-like myokine released into the circulation in response to exercise with distinct functions in regulating apoptosis, inflammation, and oxidative stress. Recent advances revealed the role of irisin as a novel therapeutic method and an important mediator of the beneficial effects of exercise in cardioprotection. Here, by using a low-dose long-term mouse DIC model, we found that the perivascular fibrosis was involved in its myocardial toxicity with the underlying mechanism of endothelial-to-mesenchymal transition (EndMT). Irisin treatment could partially reverse DOX-induced perivascular fibrosis and cardiotoxicity compared to endurance exercise. Mechanistically, DOX stimulation led to excessive accumulation of ROS, which activated the NF-κB-Snail pathway and resulted in EndMT. Besides, dysregulation of autophagy was also found in DOX-treated endothelial cells. Restoring autophagy flux could ameliorate EndMT and eliminate ROS. Irisin treatment significantly alleviated ROS accumulation, autophagy disorder, NF-κB-Snail pathway activation as well as the phenotype of EndMT by targeting uncoupling protein 2 (UCP2). Our results also initially found that irisin was mainly secreted by cardiomyocytes in the cardiac microenvironment, which was significantly reduced by DOX intervention, and had a protective effect on endothelial cells in a paracrine manner. In summary, our study indicated that DOX-induced ROS accumulation and autophagy disorders caused an EndMT in CMECs, which played a role in the perivascular fibrosis of DIC. Irisin treatment could partially reverse this phenomenon by regulating UCP2. Cardiomyocytes were the main source of irisin in the cardiac microenvironment. The current study provides a novel perspective elucidating the pathogenesis and the potential treatment of DIC.

Potential Role of Novel Myokine in Rats-Induced Metabolic Syndrome

One of the main health problems is metabolic syndrome (MetS). Its incidence elevates with age leading to a higher risk of evolving chronic diseases and cancer. Obesity and insulin resistance was considered the most vital components in its pathogenesis for a long time. This study aims to evaluate serum novel adipokine and myokine to establish the irrelation of insulin resistance and their impact on metabolic syndrome. Four groups of rats were included; the control group (C) fed with a standard diet, the second group (CI) fed on a standard diet and injected daily with irisin (100ng/ ml) till the end of the experiment. The third group (MetS group) fed on the HCHF diet for 20 weeks and served as a control group. Rats in the fourth group (MetS+I group) were fed on the HCHF diet until they become obese and diabetic, then injected daily with irisin (100ng/ ml) till the end of the experiment and served as a treated group. Serum levels of obesity and diabetes indices were significantly increased while HDL was significantly decreased in the metabolic syndrome group, but after treatment with irisin, their levels were improved. Both adropin and irisin were significantly decreased, while IL-6 was significantly increased in the same group that was enhanced after irisin treatment. In conclusion, this study demonstrated that lower irisin correlates with the increased risk of increased risk of insulin resistance and MetS. In addition, our results suggested that irisin could have a potential role in glucose metabolism. The relations among increased levels of circulating irisin, insulin resistance, and MetS prevalence may be elucidated with a physiological compensatory contrivance.

Role of Irisin in Endocrine and Metabolic Disorders—Possible New Therapeutic Agent?

Irisin is a novel hormone that provides a possible solution for the treatment of metabolic disorders. Discovered in 2012 by Boström et al., irisin very quickly became an interesting subject in medical research. Irisin has been found in cerebrospinal fluid, the cerebellum, thyroid, pineal gland, liver, pancreas, testis, spleen, adult stomach, and human fetuses. Regarding the actions of irisin, both in animals and humans, the results are contradictory but interesting. Its capability to influence adipose tissue and glycemic homeostasis may be utilized in order to treat hypothyroidism, polycystic ovary syndrome, Prader–Willi syndrome, and other endocrine and metabolic disorders. Considering its osteogenic potential, irisin might be a therapeutic choice in diseases caused by a sedentary lifestyle. New data indicate that irisin treatment may serve in the treatment of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection. Furthermore, several therapeutic agents, such as insulin, metformin, fenofibrate, exenatide, and melatonin, influence the concentrations of irisin in animal models or in humans. Nutritional factors including polyunsaturated fatty acids may also have an effect on irisin concentrations. While it may be “too good to be true,” irisin offers many opportunities for future research that would aim to find its optimal therapeutical role in endocrine and metabolic diseases.

Irisin treatment lowers levels of phosphorylated tau in the hippocampus of pre-symptomatic female but not male htau mice.

AimsIrisin is a hormone cleaved from fibronectin type‐III domain‐containing protein 5 in response to exercise and may be therapeutic in Alzheimer's disease (AD). Irisin is shown to repair damage caused by midlife cardiometabolic risk factors for AD (i.e., diabetes mellitus; hypertension), prevent neural amyloid beta aggregation and reduce neuroinflammation. However, there are no investigations of irisin's effect on AD‐associated tauopathy in the brain. This study begins to address this gap in knowledge.MethodsTransgenic htau mice that selectively develop age‐related tauopathy were treated with recombinant irisin (100 µg/kg weekly i.p.) beginning at a pre‐symptomatic age (4 months) to determine if irisin could prevent emergence of early neuropathology. One month later, mice were sacrificed to collect brain tissue and serum. Protein levels of ptau (serine 202), inflammatory cytokine tumour necrosis factor alpha (TNFα) and FNDC5 were quantified using capillary‐based western blotting (Wes).ResultsOur data show that irisin treatment significantly reduced ptau and TNFα in the hippocampus and serum of female htau mice compared to vehicle‐treated controls. Irisin treatment did not alter ptau levels in male htau hippocampus and appeared to enhance both neural and systemic TNFα levels.ConclusionsThis study provides the first evidence that enhancing the endogenous hormone irisin may be therapeutic against emerging neuropathology in a tauopathy‐selective AD model. This is important because there are currently no disease‐modifying therapeutics available for AD, and few agents in development address the multiple disease targets irisin appears to—making irisin an intriguing therapeutic candidate for further investigation.

Irisin attenuates lipopolysaccharide-induced acute lung injury by downregulating inflammatory cytokine expression through miR-199a-mediated Rad23b overexpression

AimsAcute lung injury (ALI) is a leading cause of mortality as a result of inflammatory cytokine overexpression and increased rates of apoptosis. Therapies for ALI are yet to be thoroughly investigated. Recent evidence has shown that irisin exerts protective effects against many types of pathologies. The present study aimed to determine the function of irisin in an ALI mouse model induced by lipopolysaccharide (LPS) and the corresponding underlying mechanisms at the tissue, cellular, and molecular levels. Main methodsWe assessed irisin function in A549 cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. The cell apoptosis was evaluated by flow cytometry. Western blotting and RT-PCR were used to test expression level. Animal models of ALI was established. Key findingsWe found that irisin treatment maintained lung weight, significantly reduced inflammatory cytokine expression, and alleviated lung injury by downregulating miR-199a. In LPS-stimulated cells, forced miR-199a expression downregulated Rad23b expression by targeting its 3′ untranslated region, indicating that Rad23b is a direct target of miR-199a. SignificanceThese findings reveal that irisin can alleviate ALI by inhibiting miR-199a and upregulating Rad23b expression, suggesting that irisin has clinical potential for the treatment of ALI.

Effects of irisin on lipid in mice and glycolipid metabolism in obese mice induced by diet

Objective: To explore the role of low-dose irisin in the browning of white adipose tissue (WAT) and activation of brown adipose tissue (BAT) in mice, and its effect on the metabolic function of diet induced obesity. Methods: A total of 22 C57/BLKS/J male mice fed with normal diet and 8 fed with high fat diet were separately divided into experimental and control group. The experimental group was given irisin (0.8 ng/g, 200 μl), while the control group was given the same volume (200 μl) of phosphate buffer saline every day for 14 consecutive days intraperitoneally. Food intake and body weight of mice were collected regularly every day. After intervention, the mice were killed and the changes of lipid content and activity in adipose tissue were detected by histopathology and immunohistochemistry. The effects of irisin at different concentrations (0, 20 and 40 nmol/L) on primary white adipocytes and brown adipocytes were evaluated by immunohistochemistry on uncoupling protein 1(UCP1). In order to further evaluate whether irisin has the function of improving metabolism, the changes of serum indexes and hepatic steatosis in mice fed with high-fat diet were monitored. Results: The primary white and brown adipocytes derived from mice were successfully cultured and identified in vitro. In NCD mice, the weight gain of mice with irisin was lower than that of control mice [(-0.78±0.98) vs (0.27±0.55) g]. Histopathology showed that the area of white adipocytes with irisin was smaller than controls [(14.78±8.44) vs (29.49±12.97) μm2] and the brown adipocytes were larger than controls [(0.92±0.35) vs (0.19±0.12) μm2] (both P<0.05), while the expression of UCP1 in both adipose tissues was significantly higher in irisin group. After irisin treatment, the levels of blood glucose [(7.18±0.41) vs (13.48±2.07) mmol/L, P<0.01]and cholesterol [(2.38±0.26) vs (3.89±0.93) mmol/L, P<0.05] were significantly lower than controls, and the content of lipid droplets in liver cells was less than controls [ (2.73±1.96)% vs (14.04±6.29) %, P<0.001]. Conclusions: Low dose irisin can promote the browning of WAT and activate BAT, reducing the body weight of mice by producing heat. Irisin can also effectively improve diet-induced obesity and related metabolic disorders in mice.

Irisin Has a Protective Role against Osteoporosis in Ovariectomized Rats.

The reduction in estrogen levels results in a decrease in bone density at menopause. Irisin is a myokine that modulates the benefits of exercise, which may include bone health. This study was planned to examine irisin's impact in preventing osteoporosis after ovariectomy. 4 groups of female albino rats (10 rats/group): control, sham-operated, ovariectomized (OVX-control), and OVX-irisin-treated. Serum levels of bone markers [osteocalcin (OC), bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase (TRAP), calcium (Ca++), phosphorus (P)], glucose, and insulin were being measured. Body mass index, Homeostatic Model Assessment of Insulin Resistance (HOMA-IR), dry and ash femur weight, and bone contents of Ca++ and P were investigated. The femur was examined histopathologically. The OVX-control group showed an increase in serum levels of OC, BALP, TRAP, calcium, phosphorus, BMI, glucose, insulin, and HOMA-IR (P < 0.05) and a reduction in dry and ash weight of the femur, the concentration of calcium and phosphorus content in bone ash (P < 0.05). The OVX-irisin-treated group exhibited a decrease in serum levels of OC, BALP and TRAP, calcium, phosphorus, BMI, glucose, insulin, HOMA-IR (P < 0.05), and a rise in dry and ash weight of the femur, the concentration of calcium and phosphorus in bone ash (P < 0.05). Histological examination of the distal femur diaphysis of the OVX-irisin-treated group exhibited proper bone architecture and density compared with that of the OVX-control group. It is concluded that irisin treatment in the OVX rats safeguarded the regular bone architecture and normal levels of serum bone biomarkers. Irisin may be a possible novel target in the prohibition of postmenopausal osteoporosis.

Irisin induces white adipose tissue browning in mice as assessed by magnetic resonance imaging.

This study aimed to track and evaluate the effect of low-dose irisin on the browning of white adipose tissue (WAT) in mice using magnetic resonance imaging (MRI) noninvasively in vivo. Mature white adipocytes extracted from mice were cultured, induced and characterized before being treated by irisin. The volume and fat fraction of WAT were quantified using MRI in normal chow diet and high fat mice after injection of irisin. The browning of cultured white adipocytes and WAT in mice were validated by immunohistochemistry and western blotting for uncoupling protein 1 (UCP1) and deiodinase type II (DIO2). The serum indexes were examined with high fat diet after irisin intervention. UCP1 and DIO2 in adipocytes showed increases responding to the irisin treatment. The size of white adipocytes in mice receiving irisin intervention was reduced. MRI measured volumes and fat fraction of WAT were significantly lower after Irisin treatment. Blood glucose and cholesterol levels were reduced in high fat diet mice after irisin treatment. Irisin intervention exerted browning of WAT, resulting reduction of volume and fat fraction of WAT as measured by MRI. Furthermore, it improved the condition of mice with diet-induced obesity and related metabolic disorders.

Irisin enhances osteogenic differentiation of mouse MC3T3-E1 cells via upregulating osteogenic genes.

Osteoporosis affects millions of individuals and remains a clinical challenge in terms of prevention and treatment. The present study aimed to investigate the effect of irisin on osteogenic differentiation by exposing MC3T3-E1 cells to different concentrations of irisin. Treated cells were assayed for osteoblast proliferation and osteogenic differentiation by measuring alkaline phosphatase (ALP) activity, calcium deposition, formation of mineralized nodules and the expression of osteogenic genes using reverse transcription-quantitative PCR. The proliferation of MC3T3-E1 cells was unaffected by irisin at the concentrations tested of up to 100 ng/ml (P>0.05). ALP activity and mineralized nodule formation were significantly enhanced by irisin in a dose- and time-dependent manner, indicating that irisin promotes osteoblast differentiation of MC3T3-E1 cells. The expression of osteogenic genes, including ALP, collagen I, runt-related transcription factor 2, osterix, osteopontin, osteocalcin, osteoprotegerin and estrogen receptor α, increased significantly after irisin treatment. The present study demonstrated that irisin promoted the osteogenic differentiation of MC3T3-E1 cells, possibly by upregulating the expression of osteogenic genes and markers. Therefore, irisin may be worthy of further investigation as a potential therapeutic agent for osteoporosis.

Irisin Protects Against LPS-Stressed Cardiac Damage Through Inhibiting Inflammation, Apoptosis, and Pyroptosis.

Septic cardiac dysfunction remains a clinical problem due to its high morbidity and mortality. Uncontrolled cell death and excessive inflammatory response are closely related to sepsis-induced cardiac dysfunction. Irisin has been found to play cardioprotective roles in sepsis. However, there is enough uncertainty in the mechanism of irisin-mediated cardioprotection. We hypothesized that irisin may ameliorate myocardial dysfunction via reducing cardiac apoptosis, pyroptosis, and inflammation during LPS-induced sepsis. Mice were subjected to LPS with or without irisin treatment. After stimuli of LPS, the function of myocardium was distinctly impaired, which was closely related to increased level of apoptosis (decreased expression of Bcl-2 and elevated expression of Caspase-3 and Bax), pyroptosis (increased expression of Caspase1, NLR family pyrin domain containing 3 (NLRP3), and gasdermin D) and inflammatory mediators (increased level of IL-1β, TNF-α, and IL-6). This process is consistent with increased toll-like receptor 4 (TLR4)/nuclear factor-kappa B signal, apoptotic signal, and NLRP3-mediated pyroptotic signal. Activation of apoptosis and pyroptosis enhanced the expression of proinflammatory cytokines and further exacerbated septic myocardial damage. However, irisin can inhibit the expression of TLR4 and its downstream signaling molecules and also lower the level of apoptosis and pyroptosis. Besides, similar results were also found in vitro model of LPS-induced H9c2 cardiomyocyte injury. In general, irisin suppressed inflammation, apoptosis, and pyroptosis by blocking the TLR4 and NLRP3 inflammasome signalings to mitigate myocardial dysfunction in sepsis.

Irisin Contributes to Neuroprotection by Promoting Mitochondrial Biogenesis After Experimental Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a devastating form of stroke, which poses a series of intractable challenges to clinical practice. Imbalance of mitochondrial homeostasis has been thought to be the crucial pathomechanism in early brain injury (EBI) cascade after SAH. Irisin, a protein related to metabolism and mitochondrial homeostasis, has been reported to play pivotal roles in post-stroke neuroprotection. However, whether this myokine can exert neuroprotection effects after SAH remains unknown. In the present study, we explored the protective effects of irisin and the underlying mechanisms related to mitochondrial biogenesis in a SAH animal model. Endovascular perforation was used to induce SAH, and recombinant irisin was administered intracerebroventricularly. Neurobehavioral assessments, TdT-UTP nick end labeling (TUNEL) staining, dihydroethidium (DHE) staining, immunofluorescence, western blot, and transmission electron microscopy (TEM) were performed for post-SAH assessments. We demonstrated that irisin treatment improved neurobehavioral scores, reduced neuronal apoptosis, and alleviated oxidative stress in EBI after SAH. More importantly, the administration of exogenous irisin conserved the mitochondrial morphology and promoted mitochondrial biogenesis. The protective effects of irisin were partially reversed by the mitochondrial uncoupling protein-2 (UCP-2) inhibitor. Taken together, irisin may have neuroprotective effects against SAH via improving the mitochondrial biogenesis, at least in part, through UCP-2 related targets.