Osteoprotective Effect of Cimiracemate in Glucocorticoid-Induced Osteoporosis by Osteoprotegerin/Receptor Activator of Nuclear Factor κ B/Receptor Activator of Nuclear Factor Kappa-Β Ligand Signaling

Pharmacology ◽  
2019 ◽  
Vol 103 (3-4) ◽  
pp. 163-172
Author(s):  
Zheng Ding ◽  
Huifeng Shi ◽  
Wei Yang

Objectives: Present investigation determines the protective effect of cimiracemate A against glucocorticoid-induced osteoporosis rat. Methods: Osteoporosis was induced by injecting methylprednisolone acetate (21 mg/kg) for the period of 6 weeks, and the rats were treated with cimiracemate A 5 and 10 mg/kg, p.o. 60 min after the administration of methylprednisolone acetate (21 mg/kg) for the duration of 6 weeks. Effect of cimiracemate A was observed by estimating the microarchitecture of bone and histopathological changes by micro-CT scan and light microscope. Moreover, lipid profile, mediators of inflammation, and parameters that affect bone formation were determined in the serum and western blot assay, and reverse transcription polymerase chain reaction was done for the estimation of protein expression in the bone tissues. Moreover, cytotoxic effect of cimiracemate A on bone marrow macrophages and bone marrow stromal cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Result of the investigation suggests that treatment with cimiracemate A ameliorates the microarchitecture of bone and histopathological changes in the glucocorticoid-induced osteoporosis rat. Level of lipid and mediators of inflammation was significantly reduced in the serum of cimiracemate A-treated rats than the negative control group. However, the activity of tartrate-resistant acid phosphatase and the level of collagen type I fragments in the serum were found to be reduced, and osteocalcin level was enhanced in cimiracemate A-treated rats than the negative control group. Moreover, treatment with cimiracemate A attenuates the expression of receptor activator of nuclear factor kappa-Β ligand (RANKL), receptor activator of nuclear factor κ B (RANK), and osteoprotegerin (OPG) protein in glucocorticoid-induced osteoporosis rats. Conclusion: In conclusion, our study suggests that cimiracemate A protects the glucocorticoid-induced osteoporosis by regulating the RANKL/RANK/OPG signaling pathway.

2019 ◽  
Vol 9 (9) ◽  
pp. 1254-1260
Author(s):  
Fei Wang ◽  
Hongfang Wei ◽  
Chengdong Hu ◽  
Dongfeng Li ◽  
Xiwei Huo ◽  
...  

Bone marrow mesenchymal stem cells (BMSCs) are used for bone tissue engineering. BMP-2 and autologous oxygen-releasing nano-bionic scaffolds promote bone differentiation of BMSCs. Our study intends to evaluate the role of autologous oxygen-releasing nano-bionic scaffolds combined with BMP-2-induced BMSCs in the construction of tissue engineered bone. Rat BMSCs were isolated and transfected with NC (negative control group) and BMP-2 (BMP-2 plasmid group), respectively. Healthy male SD rats were randomly and equally divided into fracture group, negative control group and the BMP-2 group which was implanted with autologous oxygen-releasing nano-bionic scaffolds to synthesize BMSCs and transfected with BMP-2 plasmids respectively followed by analysis of osteophytes growth, ALP activity, expression of BMP-2, type II collagen, Runx2 and OC by real time PCR, TGF-β1 secretion by ELISA and BMP-2 protein expression by western blot. BMSCs induced by autologous oxygen release nano-bionic scaffold combined with BMP-2 can significantly promote the increase of bone mineral density, increase the expression of Runx2 and OC, promote ALP activity, upregulate type II collagen, BMP-2 mRNA and protein, and TGF-β1 secretion compared to fracture group (P < 0.05). The BMSCs induced by autologous oxygen-releasing nanobionic scaffolds transfected with BMP-2 had a more significant effect on bone repair. Autologous oxygen-releasing nano-bionic scaffolds combined with BMP-2-induced BMSCs can promote bone healing by regulating BMP-2 and increasing osteogenesis at the bone defect.


Author(s):  
Imad A. Al-Obaidi ◽  
Nada N. Al-Shawi

Abstract At any moment, the continuous usage of medications can accompanied by DNA damage and the accumulation of such damages can cause serious consequences. Antidepressants are long-term used drugs and the incidence of their genotoxic impacts cannot be excluded. Therefore, this work was designed to investigate the possible genotoxic effects of the commonly used antidepressants (fluoxetine and amitriptyline) in adult male rats. Detection of DNA damage in individual cells was assessed by comet and micronucleus assays in three different cell populations i.e. liver, testis and bone marrow tissues of 24 swiss albino adult male rats. The animals were randomly allocated into three groups of 8 rats each: Group I - rats orally-administered distilled water via gavage tube for four weeks as a negative control. Group II - rats orally-treated with fluoxetine hydrochloride solution (7.2mg/kg/day) via gavage tube for four weeks. Group III - rats orally-treated with amitriptyline hydrochloride solution (27mg/kg/day) via gavage tube for four weeks. The results showed that both drugs (Group II and Group III) induced the same extent of DNA damage, as evidenced by a significantly higher DNA fragmentation in liver and testis tissues with increased frequencies of micronuclei formation in bone marrow tissues as compared with the negative control (Group I). These findings indicates that both Fluoxetine and Amitriptyline have genotoxic potentials and can induce the same extent of cytogenetic damage in rats. Special precautions and medical supervision should be taken in consideration with their uses.


Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5420-5420
Author(s):  
Jianyu Weng ◽  
Xin Huang ◽  
Xiaomei Chen ◽  
Qiong Luo ◽  
Jianhua Su ◽  
...  

Abstract Background: Our preliminary research found that STAT3, IL-17A, and IL-21 expressed in cGVHD patients. So, we provide the blocking STAT3 signal to the induction of Treg cells differentiation, and to provide experimental basis on new targets of cGVHD immunotherapy. Methods: 1. Mice spleen CD4+ CD62L+ naiveT cells were separated by immune magnetic bead and then activated for 72 h. After 96 h infection with STAT3-shRNA and negative control lentivirus, the Th subgroup proportion were measured by flow cytometry. Th related cytokines levels test by Luminex. Real time quantitative PCR was to detect STAT3 and Th subgroup related transcription factor mRNA levels. CD117+ mouse bone marrow stem progenitor cells were sorted by flow cytometry, and transfected by STAT3-shRNA. Inhibition of STAT3 gene in mRNA level was measured at 96 h. Cell proliferation activity was test with CCK8 kit, and cell apoptosis rate determined by flow cytometry. Differentiation of CD117+ cells was induced by 2.2% of methyl cellulose and different cytokines. 2. BALB/c female mice, after the linear accelerator 700cGy of whole body irradiation, accepted miHA mismatched male B10. D2 mice bone marrow cells and spleen cells (8 x 106, 1:1). Randomly assigned 6 mice of cGVHD clinical score of 0.6 or above to each group. After STAT3-shRNA or negative control lentivirus treatment, the observe end point was 58th day after transplantation. The clinical and pathologic scores compared. Th17 and Treg cells measured by flow cytometry. Th related cytokines measured by Luminex. Purpose genes in blood and protein expression levels in target organs were found by Q-PCR and western blot test, respectively. Results: 1. The Th17 / Treg ratio of shRNA group was significantly decreased than that in the NC group (P < 0.05). Except for the Foxp3 gene, other purpose genes, including T-bet, Gata3, RORγt, TGFβ, Notch1, and Jagged2 mRNA levelsin interference group were cut. GM-CSF, IFN-gamma, beta, IL- 3, IL-2, IL-4, IL-6, TNF alpha, IL-17, IL-22a, IL-27, and IL-9 factor expression levels were significant difference between shRNA and negative control group (P < 0.05). There was no significant difference of cell proliferation activity, early apoptosis rate, and differentiation ability in STAT3-shRNA treated CD117+ bone marrow, compared with negative control group and blank control group (P > 0.05). 2. After 50th day, shRNA treatment group appear hair recovery, energy recovery, weight gain, shortness of breath better, mean of cGVHD score decreased. At the 58th day, clinical scores of cGVHD between shRNA treatment group and the negative control group overall mean difference was statistically significant (P < 0.05). cGVHD pathological score of lungs in shRNA treat group reduced (P < 0.05). STAT3mRNA levels in peripheral blood, phosphorylated STAT3, and STAT3 expression level of lung declined than control groups. The proportion of Th17 / Treg cells of spleen was significant reduced in shRNA group, compared with negative control group (P < 0.05). Conclusion: 1. STAT3 knocking down in naïve CD4 + Th cells induced the increased Treg cells, and the decreased Th17 cells. IL-2 confirmed to promote the growth of Treg cells. It speculated that blocking STAT3 might bring Th9 cells differentiation. STAT3 blocking in CD117+ stem progenitor cells have no significant effect on the proliferation, apoptosis and differentiation, validation the safety of STAT3-shRNA. 2. STAT3-shRNA treatment cGVHD mice in vivo achieved curative effect. The main target organs was the lung, which might be closely related to the fall in the proportion of Th17 /Treg. STAT3 may be used as a new target for immunotherapy of cGVHD. Acknowledgment The project was sponsored by grants from National Natural Science Foundation of China (No. 30972790; No.81270648; No.81370665; No.81300446), Provincial Natural Science Foundation of Guangdong (No. S2012010009560), Provincial Science and Technology Planning Project of Guangdong (No.2013B021800186; No.2013B021800201), and Science and Technology Planning Project of Guangzhou (No. 201400000003-4, 201400000003-1). Disclosures No relevant conflicts of interest to declare.


BioScience ◽  
2019 ◽  
Vol 3 (1) ◽  
pp. 85
Author(s):  
Rina Priastini Susilowati

In this study, mice testicular degeneration was influenced by exposure to the mosquito mat made from transfluthrin 3000 ppm and permot leaf mosquito mat which were evaluated based on observations on testicular histopathological changes and testosterone level. Fifteen Balb C male mice aged 2-3 months with a body weight of 25-30 g were divided into five groups, namely A, B, C, D, and E, with each group consisting of 3 replications. Group A was the negative control group (without exposure), group B was the exposure group for the mosquito mat made from transfluthrin 3000 ppm, group C, D, and E were the exposure groups of mosquito mat with permot leaves doses of 1000 ppm, 2000 ppm and 3000 ppm. Each exposure group was given treatment 8 hours per day from 18.00 - 04.00 for 3 months. The results showed a significant decrease in testicular weight between the control group and the treatment group. There were also significant differences in the testosterone level observed in the treatment group, as well as observations on the spermatogenic index (IS). For histopathological observation of testicular mice, there was a reduction in epithelial cell spermatogenic and diameter of seminiferous tubule. The results of the Kruskall Wallis test showed a significant difference between the control group and the treatment group of the mosquito mat made from transflutrin 3000 ppm. The conclusion is that exposure to mosquito mat made of permot peaves up to a 3000 ppm dose is safe to use and can kill the Aedes aegypti mosquito effectively.


2021 ◽  
Vol 43 (1) ◽  
Author(s):  
Zohre Khodabandeh ◽  
Mahmoud Etebari ◽  
Mehdi Aliomrani

Abstract Background and aim Approximately, 2 million tonnes of pesticides are utilized annually worldwide. Phosalone (Pln), an organophosphorus pesticide, acts as an insecticide and acaricide to control pests of crops such as nuts, citrus fruits, pomegranates, stone fruits, grapes, potatoes, and artichokes. The purpose of this study was to evaluate the possible genotoxic effects following exposure to Pln in the cells derived from mouse red bone marrow. Materials and methods Sixty mice were divided into 6 groups including cyclophosphamide (40 mg/kg, IP) and Pln (6, 12, 20, and 40 mg/kg) exposure by gavage. After 1 and 5 days of exposure, animals were euthanized and the genotoxicity assays were done on bone marrow extracted cells. Results Comet assay shows a time and dose-dependent toxicity which further DNA degradation is observed after 5-day exposure (p < 0.05). Also, Pln significantly increased the MnPCE/PCE ratio after 12 and 20 mg/kg administration while no significant difference was reported between the doses of 6 and 40 mg/kg BW with the negative control group. Conclusion Our results suggested a serious concern about its potential effects on biological life and related disease inductions. However further studies need to confirm the exact mechanism of Pln genotoxicity and the cause of diverse response of its activity at 40 mg/kg. This study also showed that increasing the dose of Pln reduces the MnNCE/Total cells ratio, which may indicate the possibility of bone marrow suppression. All of the above results emphasize the need to seriously limit the use of this compound as an agricultural pesticide.


Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 318-318
Author(s):  
Xin Guan ◽  
Meng Qin ◽  
Bin Shen ◽  
Yu Zhang ◽  
Wenhong Jiang ◽  
...  

Abstract Background and Objectives: Due to platelet shortage, megakaryocytes have been regarded as an effective substitute to the alleviation of frequent thrombocytopenia after stem cell transplantation. However, ex vivo expansion of megakaryoblasts and their subsequent differentiation into mature megakaryocytes for clinical applications remains a challenge. Here, we describe the development of a two-stage culture system for producing megakaryocytes from cord blood CD34+ cells. Design and Methods: Firstly, we expanded CD34+ hematopoietic progenitor/stem cells for 6 days in a serum-free culture system (IMDM basal medium with the addition of biotin, putrescine, insulin, human serum albumin, selenium, and some other nutrients) supplemented with stem cell factor (SCF), Flt-3 ligand (FL), thrombopoietin (TPO), interleukin 3 (IL-3), low density lipoprotein (LDL), StemRegenin 1 (SR1), and DMSO. CD34+ cells expansion was monitored by flow cytometric analysis of cell surface markers coupled with cell counting. Subsequently, these expanded cells were induced toward the megakaryocytic lineage for additional 7 days in the same serum-free medium as above supplemented with SCF, TPO, IL-3, IL-6, IL-11, granulocyte-macrophage colony-stimulating factor (GM-CSF), and LDL. Megakaryocytes were detected by flow cytometry using antibodies against specific cell surface markers including CD41a and CD42b. Differentiated megakaryocytes were also confirmed by morphological criteria such as cell size and DNA polyploidy. To functionally evaluate induced megakaryocytes, these cells were transplanted into sublethally irradiated NOD/SCID mice. Viability and cell being of these mice were monitored after injection. Mice of the negative control group (n=3) were injected with saline. In the experimental group (n=6), each mouse was injected with 1.0×107 cells from the second stage of culture. Results: After the first stage culture, proliferation folds of total cells and CD34+ cells were 85.65±7.03 and 62.91±4.36, respectively. The calculated yield from each CD34+ cell was between 1.0×104 to 1.5×104 CD41+ megakaryocytes with a purity of CD41+ and CD42+ cells reaching 93.7%±2.8% and 80.3%±5.8%, respectively. Differentiated cells were morphologically discernible as they were much larger than starting CD34+ cells with apparent lobular nuclei and numerous α-granules. In addition, about 32.67%±7.43% of induced megakaryocytes exhibited 4N or larger DNA content (4N 17.6%±4.12%; 8N 10.93%±2.48%; >8N 3.23%±1.34%). In mouse studies, samples collected from the negative control group contained no cells positive for human CD41a and CD42b markers. In the experimental group, human platelets were detected in mouse peripheral blood 3 days post-transplantation. At day 14 post-transplantation, the percentage of platelets derived from injected human megakaryocytes reached 13.6%±6.2%. Human megakaryocytes were also detected in mouse bone marrow 7 days post-transplantation, peaking at day 14 (~2.38% of total bone marrow megakaryocytes). Conclusions: We have established a stem cell expansion and differentiation platform that can be adapted to large-scale production of mature megakaryocytes from umbilical cord blood cells. Significantly, induced megakaryocytes are capable of engrafting in mouse bone marrow and producing platelets after transplantation into irradiated NOD/SCID mice. Therefore, our experimental platform is capable of producing a sufficient number of functional megakaryocytes for various clinical applications in the future. Disclosures Qin: Biopharmagen corp: Employment. Jiang:Biopharmagen.corp: Employment. Ren:Biopharmagen corp: Employment. Jiang:Biopharmagen.corp: Employment.


2021 ◽  
pp. 1354-1362
Author(s):  
Ali Arbaga ◽  
Amanallah El-Bahrawy ◽  
Ahmed Elsify ◽  
Hadeer Khaled ◽  
Hany Youssef Hassan ◽  
...  

Background and Aim: Pyoderma is common in dogs, and its treatment requires a novel medication rather than antibiotic therapy. This study aimed to determine the biochemical and histopathological changes associated with the topical application of Aloe vera 20% and 40% ointments, compared with gentamicin 0.1% ointment, in dogs suffering from Staphylococcus aureus pyoderma. Materials and Methods: Serum and skin samples were collected from a negative control group before inducing pyoderma and from other subdivided groups on the 3rd, 7th, 10th, and 14th days post-inoculation for biochemical and histopathology examination. Results: Serum aspartate aminotransferase, alanine aminotransferase (ALT), urea, and creatinine concentrations were higher in the positive control dogs on the 3rd day without treatment (DWT) compared with the negative control dogs (p<0.05). Compared with the healthy control dogs, serum zinc concentrations were lower in the positive control group on the 3rd, 7th, and 10th DWT and in dogs treated with A. vera 20% and gentamicin 0.1% ointments on the 3rd and 7th days post-treatment (p<0.05). Grossly, skin had erythema, pruritus, and pus-filled pustules of the untreated group. Microscopically, skin showed epidermal necrosis and edema, dermal collagen necrosis, and severe neutrophilic infiltration. Conclusion: Compared with A. vera 20% and gentamicin 0.1% ointments, the topical application of A. vera 40% ointmentinduced quicker skin healing and decreased the inflammatory changes caused by S. aureus inoculation, based on biochemical and histopathological changes reflective of its curative efficiency. A. vera 40% ointment may be a suitable alternative to antibiotics for the treatment of staphylococcal pyoderma in dogs.


2018 ◽  
Author(s):  
Linyuan Wang ◽  
Fang Xiao ◽  
Lei Yu ◽  
Hang Su ◽  
Yueqin Yang ◽  
...  

AbstractObjectiveBone marrow cells encounter various chemical and mechanical stimuli from the internal environment. In vivo, fluid shear stress (FSS) is one of the primary mechanical stimuli that affect bone marrow-derived mesenchymal stem cells (BMCs) activity. Since various cases of FSS influence BMCs activity differently, the purpose of this study is to determine how BMC activity in osteogenesis and osteoclastgenesis is affected by stable and unstable changing FSS.MethodBMCs samples from the femur of the mouse, divided in to three groups: stimulate by stable changing FSS, unstable changing FSS, and no FSS. RT-PCR would be applied to detect OPG, RANKL, ALP, OCN, RUNX2, and RANK of all the samples at the 3rd day. Alizarin red staining and TRAP staining would be applied to test all the samples at the 6th day.ResultsThe S group samples showed the lowest level in RANKL mRNA and showed the highest level in OPG mRNA. RANKL/OPG mRNA in the S group was the smallest during three groups. Comparing with the C group samples, RUNX2 mRNA in the S group was increased significantly. RANK mRNA in the C group was three times more than S group. S group had the largest area of mineralized nodules, and largest number of area ≥100µm, ≥500µm2 or ≥1000µm2. The area of positive TRAP stain in S group was the smallest among three groups.ConclusionStable changing FSS significantly increases osteogenesis relating to OPG-RANKL-RANK pathway. Compared with unstable changing FSS, stable changing FSS afford a more appropriate stimulation to osteogenesis.AbbreviationsALPAlkaline phosphataseBMCsBone marrow-derived mesenchymal stem cellsCx43Connexin 43C3aComplement Component 3aJNKc-Jun N-terminal kinaseERKExtracellular regulated protein kinasesERK5Extracellular-signal-regulated kinaseFSSFluid shear stressFasLFirst apoptosis signal LigandGSK3βGlycogen synthase kinase 3βLPALysophosphatidic acidMSCsMesenchymal stem cellsM-CSFMacrophage colony-stimulating factorMCP-1Monocyte chemoattractant protein-1RANKReceptor Activator of Nuclear Factor κ BRANKLReceptor activator of nuclear factor κ B ligandRUNX2Runt-related transcription factor 2P2Y2RReceptor of P2Y2LGR4Repeat-containing G-protein-coupled receptor 4SemaSemaphorinsTRAPTartrate-Resistant Acid AhosphataseTRPM7Transient receptor potential cation channel subfamily M member 7TRAFsTumornecrosisfactorreceptor associatedfactorsOPGOsteoprotegerinOCNOsteocalcinOPNOsteopontin


2014 ◽  
Vol 92 (9) ◽  
pp. 733-743 ◽  
Author(s):  
Mohamed Sarhan ◽  
Hanaa El Serougy ◽  
Abdelaziz M. Hussein ◽  
Mohamed El-dosoky ◽  
Mohamed A. Sobh ◽  
...  

Objectives: To study the effects of bone-marrow-derived mesenchymal stem cells (BM-MSCs) on adriamycin (ADR)-induced chronic nephropathy in rats. Methods: 60 male Sprague–Dawley rats were distributed among 3 groups (20 rats each): (i) the negative control group, which was normal rats that received saline (vehicle); (ii) the positive control (ADR) group, which was rats that received 2 intravenous injections of ADR into the penile vein at 14 day intervals without treatment, and (iii) the MSC group, which were rats treated as for the ADR group that were also given 2 intravenous injections of MSCs (5 days after each ADR injection). Results: ADR caused a significant reduction in animal body mass, survival rate, hemoglobin (Hb) content, serum albumin, and renal GSH, and significantly increased serum levels of triglycerides, cholesterol, urinary protein excretion and kidney injury molecule-1 (KIM-1), renal MDA, as well as caspase-3 expression and glomerular and tubulointerstitial damage compared with the negative control group. MSC treatment failed to improve animal survival rate, body mass, Hb level, proteinuria, or hypoalbuminemia; however, it mildly improved the serum BUN, hyperlipidemia, caspase-3 expression, urinary levels of KIM-1, renal oxidative stress markers, and glomerular and tubulointerstitial damage score. Conclusion: administration of BM-MSCs during induction of ADR nephropathy provides partial protection, which could be due to improvements in the levels of of endogenous antioxidants, reduction of apoptosis, and maintenance of the integrity of the glomerular membrane.


2014 ◽  
Vol 11 (4) ◽  
pp. 1448-1454
Author(s):  
Baghdad Science Journal

The aim of this study to conduct the effects of fimbrial and lipopolysacchride (LPS) immunization is on the pathohistological changes in rabbits, Fifteen rabbits of both sexes (Weight 1500-2000 gm) divided into three groups (5 animals of each group). The first group was immunized by 1ml (200µg /animal) of fimbrial subcutaneously the second group gave 1 ml ( 200 µg /animal) LPS while the third group was left as negative control group that injected 1 ml phosphate buffer control subcutaneously. First and second groups recived the same dose after two weeks give as booster dose. All animals challenged after 5 weeks of immunization by5X107CFU/ml Proteus vulgaris intra peritoneally .After 7 days from challenge all the animals, sacrificed for histopathological examination . The results showed that the fimbrial group had a severe infiltrations of mononucleart cells in liver and kidney ,but there was no clear histopathological changes observed in the spleen compared with lipopolysaccharide and control group . Also this group showed a slight mononuclear cells infiltration in lamina properia of intestine ;while the lipopolysaccharide group showed hypertrophy of epithelial cells with a mild mononuclear cells infiltration in lamina properia as compared with the control group that showed presence of mononuclear cells aggregation in the lamina properia .Our conclusion That the defense of animals against Proteus vulgaris more efficient in fimbrial antigen than lipopolysaccharide antigen by decrease the pathological effects of this bacteria.


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