Satellite Cells Are Activated In A Rat Model of Radiation-Induced Muscle Fibrosis

Abstract Background: Radiation-induced muscle fibrosis is a long-term side effect of radiotherapy that significantly affect the quality of life and even reduces the survival of cancer patients. We have demonstrated that radiation induces satellite cell (SC) activation at the molecular level; however, cellular evidence in a rat model of radiation-induced muscle fibrosis was lacking. In this study, we evaluated SC activation in vivo and investigated whether radiation affects the proliferation and differentiation potential of SCs in vitro. Methods: For in vivo studies, Sprague-Dawley rats were randomly divided into six groups (n = 6 per group): a non-irradiated control group and 90 Gy-1 w, 90 Gy-2 w, 90 Gy-4 w, 90 Gy-12 w, and 90 Gy-24 w groups.Left groin area of the rats received a single dose of irradiation and rectus femoris tissues were collected in the indicated weeks. Fibrosis, apoptosis, and autophagy were evaluated by Masson’s trichrome staining, TUNEL staining, and electron microscopy, respectively. SC activation and central nuclear muscle fibers were evaluated by immunofluorescence staining and hematoxylin and eosin staining. IL-1β concentrations in serum and irradiated muscle tissue samples were determined by ELISA. For in vitro studies, SCs were isolated from rats with radiation-induced muscle fibrosis and their proliferation and differentiation were evaluated by immunofluorescence staining.Results: In vivo, fibrosis increased over time following irradiation. Apoptosis and autophagy levels, IL-1β concentrations in serum and irradiated skin tissues, and the numbers of SCs and central nuclear muscle fibers were increased in the irradiated groups when compared with control group. In vitro, cultured SCs from irradiated muscle were positive for proliferation marker Pax7, and differentiated SCs were positive for myogenic differentiation marker MyHC.Conclusion: This study provided cellular evidence of SC activation and proliferation in rats with radiation-induced muscle fibrosis. Radiation does not affect the proliferation and differentiation potential of SCs in vitro.

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Effects of Naringin on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells In Vitro and In Vivo

Stem Cells International ◽

10.1155/2015/758706 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 19

Author(s):

Lihua Yin ◽

Wenxiao Cheng ◽

Zishun Qin ◽

Hongdou Yu ◽

Zhanhai Yu ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Periodontal Ligament ◽

Trabecular Structure ◽

Control Group ◽

Periodontal Ligament Stem Cells ◽

Expression Levels ◽

Proliferation And Differentiation

This study is to explore the osteogenesis potential of the human periodontal ligament stem cells (hPDLSCs) induced by naringin in vitro and in vitro. The results confirmed that 1 μM naringin performs the best effect and a collection of bone-related genes (RUNX2,COL1A2, OPN, and OCN) had significantly higher expression levels compared to the control group. Furthermore, a typical trabecular structure was observed in vivo, surrounded by a large amount of osteoblasts. These results demonstrated that naringin, at a concentration of 1 μM, can efficiently promote the proliferation and differentiation of hPDLSCs both in vitro and in vivo.

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Developmental aspects of adipose tissue in GH receptor and prolactin receptor gene disrupted mice: site-specific effects upon proliferation, differentiation and hormone sensitivity

Journal of Endocrinology ◽

10.1677/joe.1.06939 ◽

2006 ◽

Vol 191 (1) ◽

pp. 101-111 ◽

Cited By ~ 39

Author(s):

David J Flint ◽

Nadine Binart ◽

Stephanie Boumard ◽

John J Kopchick ◽

Paul Kelly

Keyword(s):

Adipose Tissue ◽

Receptor Gene ◽

Metabolic Effects ◽

Wild Type ◽

Extrinsic Factors ◽

Differentiation Potential ◽

Site Specific ◽

Proliferation And Differentiation

Direct metabolic effects of GH on adipose tissue are well established, but effects of prolactin (PRL) have been more controversial. Recent studies have demonstrated PRL receptors on adipocytes and effects of PRL on adipose tissue in vitro. The role of GH in adipocyte proliferation and differentiation is also controversial, since GH stimulates adipocyte differentiation in cell lines, whereas it stimulates proliferation but inhibits differentiation of adipocytes in primary cell culture. Using female gene disrupted (ko) mice, we showed that absence of PRL receptors (PRLRko) impaired development of both internal and s.c. adipose tissue, due to reduced numbers of adipocytes, an effect differing from that of reduced food intake, where cell volume is decreased. In contrast, GHRko mice exhibited major decreases in the number of internal adipocytes, whereas s.c. adipocyte numbers were increased, even though body weight was decreased by 40–50%. The changes in adipose tissue in PRLRko mice appeared to be entirely due to extrinsic factors since preadipocytes proliferated and differentiated in similar fashion to wild-type animals in vitro and their response to insulin and isoproterenol was similar to wild-type animals. This contrasted with GHRko mice, where s.c. adipocytes proliferated, differentiated, and responded to hormones in identical fashion to controls, whereas parametrial adipocytes exhibited markedly depressed proliferation and differentiation potential and failed to respond to insulin or noradrenaline. Our results provide in vivo evidence that both GH and PRL stimulate differentiation of adipocytes but that the effects of GH are site specific and induce intrinsic changes in the precursor population, which are retained in vitro.

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P11.22 Radiotherapy effects in GBM Rat model CNS1

Neuro-Oncology ◽

10.1093/neuonc/noz126.168 ◽

2019 ◽

Vol 21 (Supplement_3) ◽

pp. iii47-iii47

Author(s):

O Furman ◽

D Daniels ◽

D Guez ◽

D Last ◽

S Sharabi ◽

...

Keyword(s):

Rat Model ◽

Tumor Size ◽

Growth Arrest ◽

Control Group ◽

Lewis Rats ◽

Post Irradiation ◽

Initial Tumor ◽

Initial Tumor Size

Abstract BACKGROUND CNS1 is a syngeneic glioma model in Lewis Rats. It is an aggressive infiltrating tumor cell line that mimics important aspects of human GBM such as rapid growth, dispersal along blood vessels and white matter, pseudopallisading cells with features of hemorrhage and necrosis. CNS1 tumors are infiltrated with macrophages and T-cells, and were studied in the context of immunotherapy and gene therapy, extracellular matrix and invasion, but CNS1 response to radiation has not yet been described. If we wish to combine novel immune-based therapies with existing GBM protocols that include radiation and chemotherapy, we will need models that respond to these protocols. As a first step in this direction, we sought to describe CNS1 response to radiation in vitro and in vivo. MATERIAL AND METHODS In vitro, survival of irradiated CNS1 cells was assessed with clonogenic assay. Radiation varied in dose from 0 to 10 Gy and was delivered via Kimtron Polaris X-ray generator. In vivo, male Lewis rats were intra-cranially inoculated with 0.5*106 CNS1 tumor cells and monitored for survival. Treated rats (N=6) were subjected to a single 20Gy whole-head radiation treatment under full anesthesia, delivered five days post-inoculation. Control rats (N=5) were anesthetized but not irradiated. Tumor size was monitored using contrast enhanced T1-weighted MRI in both treated and control rats at several time points (4, 6, 11, 18 and 32 days post tumor inoculation). RESULTS CNS1 cells are sensitive to radiation in vitro, as cell survival decreased after exposure to increasing amounts of radiation. In vivo, while initial tumor size did not significantly differ between groups, rats treated with radiation survived significantly longer than control rats (23.8 ± 5.0 days vs. 11 ± 4.1 days, p<0.005). Growth arrest following irradiation in vivo was not detected 1d after treatment but was observed 6d post-irradiation. Growth arrest was recorded in half of the treated rats, showing no increase in tumor size (N=2) or reduction in tumor volume (N=1) relative to 1d post-irradiation. Tumor growth rates were lower in all irradiated rats relative to control rats. Survival time was negatively correlated with initial tumor size in the control group but not in the treatment group. CONCLUSION CNS1 rat model of GBM is a valid model of radiotherapy effects on GBM tumors. Further studies combining radiation and chemotherapy are the next step. Support for this work was provided by Israel Cancer Association.

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Bioengineered Trachea with Fibroblasts in a Rabbit Model

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348941011901109 ◽

2010 ◽

Vol 119 (11) ◽

pp. 796-804 ◽

Cited By ~ 1

Author(s):

Wataru Okano ◽

Yukio Nomoto ◽

Ikuo Wada ◽

Ken Kobayashi ◽

Masao Miyake ◽

...

Keyword(s):

Rabbit Model ◽

Control Group ◽

Epithelial Regeneration ◽

Proliferation And Differentiation ◽

Accelerated Proliferation ◽

Group A ◽

Stratified Squamous Epithelium ◽

Immunohistochemical Studies

Objectives: Although our group has had mostly successful results with clinical application of a tracheal prosthesis, delayed epithelial regeneration remains a problem. In our previous studies using rats, it was demonstrated that tracheal fibroblasts accelerated proliferation and differentiation of the tracheal epithelium in vitro and in vivo. The purpose of this study was to evaluate the effects of fibroblasts on epithelial regeneration in larger tracheal defects in rabbits. Methods: We developed a bioengineered scaffold, the luminal surface of which was coated with fibroblasts. This scaffold was implanted into tracheal defects in 12 rabbits (bioengineered group), and scaffolds without fibroblasts were implanted in 12 rabbits (control group). The regenerated epithelium was histologically examined by light microscopy, scanning electron microscopy, and immunohistochemical studies. Results: In the bioengineered group, a stratified squamous epithelium was observed on the surface 7 days after transplantation. However, in the control group, the scaffolds were exposed. Fourteen days after implantation, a columnar ciliated epithelium was observed in the bioengineered group. The average thickness of the regenerated epithelium in the bioengineered group was significantly greater than that in the control group. Conclusions: This study indicated that fibroblasts had a stimulatory effect that hastened regeneration of the epithelium in large tracheal defects.

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The Effects of Photobiomodulation on Bone Defect Repairing in a Diabetic Rat Model

International Journal of Molecular Sciences ◽

10.3390/ijms222011026 ◽

2021 ◽

Vol 22 (20) ◽

pp. 11026

Author(s):

Ji-Hua Lee ◽

Su-Chii Kong ◽

Chia-Hsin Chen ◽

Ying-Chun Lin ◽

Kun-Tsung Lee ◽

...

Keyword(s):

Osteogenic Differentiation ◽

Bone Defect ◽

Rat Model ◽

In Vitro Study ◽

Diabetic Rat ◽

Control Group ◽

Calvarial Defect ◽

Diabetic Rat Model

The purpose of this study is to examine the prospective therapeutic effects of photobiomodulation on the healing of bone defects in diabetic mellitus (DM) using rat models to provide basic knowledge of photobiomodulation therapy (PBMT) during bone defect repair. For in vitro study, an Alizzarin red stain assay was used to evaluate the effect of PBMT on osteogenic differentiation. For in vivo study, micro-computed tomography (microCT) scan, H&E and IHC stain analysis were used to investigate the effect of PBMT on the healing of the experimental calvarial defect (3 mm in diameter) of a diabetic rat model. For in vitro study, the high glucose groups showed lower osteogenic differentiation in both irradiated and non-irradiated with PBMT when compared to the control groups. With the PBMT, all groups (control, osmotic control and high glucose) showed higher osteogenic differentiation when compared to the non-irradiated groups. For in vivo study, the hyperglycemic group showed significantly lower bone regeneration when compared to the control group. With the PBMT, the volume of bone regeneration was increasing and back to the similar level of the control group. The treatment of PBMT in 660 nm could improve the bone defect healing on a diabetic rat calvarial defect model.

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Increased Trimethylamine N-oxide Contributes to Metabolic dysfunction in a Rat Model of PCOS and Decreases Mitochondrial function

10.21203/rs.3.rs-122457/v1 ◽

2020 ◽

Author(s):

Huang Jiayu ◽

Liu Jiaying ◽

Zhang Hanke ◽

Li Yajie ◽

Minuo Yin ◽

...

Keyword(s):

Rat Model ◽

Mitochondrial Function ◽

Group Treatment ◽

Intestinal Flora ◽

Control Group ◽

Metabolic Dysfunction ◽

Childbearing Age ◽

The Relationship

Abstract Polycystic ovarian syndrome (PCOS) is a common endocrine disease in adolescents and women of childbearing age, also a common cause of female infertility. In recent years, studies have found that the occurrence of PCOS is related to changes in the intestinal flora. Trimethylamine N-oxide (TMAO) is an organic compound produced by intestinal microorganisms. However, the relationship between TMAO and PCOS remain mostly unexplored. The effects of TMAO on PCOS were assessed in vitro and in vivo. In a PCOS rat model, plasma TMAO, hormone and PI3K signaling levels were examined. In the process of in vitro maturation (IVM), immunofluorescence and confocal microscopy were used to detect the influence of adding different TMAO concentrations to the culture medium on oocytes. The fasting insulin (FINS), HOMA-IR, luteinizing hormone (LH), LH/follicle-stimulating hormone (FSH) and plasma TMAO levels of the PCOS rat group were significantly higher than those of the control group. Treatment with the TMAO inhibitor 3,3-dimethyl-1-butanol (DMB) alleviated metabolic disorder in PCOS rats. In PCOS rats, DMB improved the PI3K/Akt-related signaling pathway compared to no treatment. In IVM, the mitochondria of oocytes in the TMAO groups were aggregated and distributed, and mitochondrial membrane potential and ATP content were decreased. Apoptosis was more severe in the TMAO group than in the control group. TMAO worsened metabolic dysfunction in a rat model of PCOS and decreased the mitochondrial function of oocytes in the process of IVM.

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Topically Applied Bacteriophage to Control Multi-Drug Resistant Klebsiella pneumoniae Infected Wound in a Rat Model

Antibiotics ◽

10.3390/antibiotics10091048 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1048

Author(s):

Mohamed S. Fayez ◽

Toka A. Hakim ◽

Mona M. Agwa ◽

Mohamed Abdelmoteleb ◽

Rania G. Aly ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Rat Model ◽

Bacterial Infections ◽

Control Group ◽

Lytic Phage ◽

Drug Resistant ◽

Infected Wound ◽

Specific Phage

(Background): Multi-drug-resistant Klebsiella pneumoniae (MDR-KP) has steadily grown beyond antibiotic control. Wound infection kills many patients each year, due to the entry of multi-drug resistant (MDR) bacterial pathogens into the skin gaps. However, a bacteriophage (phage) is considered to be a potential antibiotic alternative for treating bacterial infections. This research aims at isolating and characterizing a specific phage and evaluate its topical activity against MDR-KP isolated from infected wounds. (Methods): A lytic phage ZCKP8 was isolated by using a clinical isolate KP/15 as a host strain then characterized. Additionally, phage was assessed for its in vitro host range, temperature, ultraviolet (UV), and pH sensitivity. The therapeutic efficiency of phage suspension and a phage-impeded gel vehicle were assessed in vivo against a K. pneumoniae infected wound on a rat model. (Result): The phage produced a clear plaque and was classified as Siphoviridae. The phage inhibited KP/15 growth in vitro in a dose-dependent pattern and it was found to resist high temperature (˂70 °C) and was primarily active at pH 5; moreover, it showed UV stability for 45 min. Phage-treated K. pneumoniae inoculated wounds showed the highest healing efficiency by lowering the infection. The quality of the regenerated skin was evidenced via histological examination compared to the untreated control group. (Conclusions): This research represents the evidence of effective phage therapy against MDR-KP.

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Therapeutic Effects of Human Urine-Derived Stem Cells in a Rat Model of Cisplatin-Induced Acute Kidney Injury In Vivo and In Vitro

Stem Cells International ◽

10.1155/2019/8035076 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 3

Author(s):

Bishao Sun ◽

Xing Luo ◽

Chengfei Yang ◽

Peilin Liu ◽

Yang Yang ◽

...

Keyword(s):

Stem Cells ◽

Acute Kidney Injury ◽

Rat Model ◽

Kidney Injury ◽

Ultrastructural Changes ◽

Great Promise ◽

Therapeutic Effects ◽

Differentiation Potential

Acute kidney injury (AKI) is an extremely dangerous clinical syndrome with high morbidity and mortality. Stem cell-based therapies have shown great promise for AKI treatment. Urine-derived stem cells (USCs) are a novel cell source in tissue engineering and cell therapy which provide advantages of simple, noninvasive, and low-cost harvest methods, efficient proliferation, and multi-differentiation potential. Here, we described the therapeutic effects of USCs in a rat model of cisplatin-induced AKI as a novel therapy. In vivo, the intravenous administration of USCs alleviated the renal functional damage in AKI rats, for the levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were significantly decreased. The USCs-treated group also exhibited improved histological and ultrastructural changes, promoted proliferation, and inhibited apoptosis in renal tissues. After the USC therapy, the expression levels of proinflammatory cytokines (TNF-α and IL-6) and apoptosis-related proteins (BAX and cleaved caspase-3) were downregulated. In addition, the presence of a few GFP-labeled USCs was confirmed in rat renal tissues. In vitro, rat tubular epithelial (NRK-52E) cells were incubated with cisplatin to induce cell damage and then cocultured with USCs. After coculture with USCs, the cisplatin-induced NRK-52E cells showed higher cell viability and a lower apoptosis ratio than those of the control group, and cell cycle arrest was improved. In conclusion, our results demonstrated that USC therapy significantly improved the renal function and histological damage, inhibited the inflammation and apoptosis processes in the kidney, and promoted tubular epithelial proliferation. Our study exhibited the potential of USCs in the treatment of AKI, representing a new clinical therapeutic strategy.

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In Vitro and In Vivo Antioxidative Activity against Radiation-Induced Damage and the Systematic Chemical Components of Different Extracts of Lagotis brevituba Maxim

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/9726431 ◽

2020 ◽

Vol 2020 ◽

pp. 1-14

Author(s):

Dan Zhang ◽

Lihong Tan ◽

Ling Yao ◽

Wei Tao ◽

Ruixue Gong ◽

...

Keyword(s):

Antioxidant Activity ◽

Antioxidant Activities ◽

Whole Body ◽

Chemical Components ◽

Control Group ◽

Dependent Manner ◽

Antioxidant Assays ◽

Radiation Induced

Lagotis brevituba Maxim is a perennial species distributed in the highlands of China, which has been used for more than 2000 years as a traditional Tibetan medicinal plant. However, no attention has been paid to the antioxidant activities of Lagotis brevituba Maxim in vitro or in vivo. Thus, this study aimed to evaluate the in vitro and in vivo antioxidant activity of Lagotis brevituba Maxim against radiation-induced damage as well as the systematic chemical components. To explore the relationship between the antioxidant activity and extraction solvent, Lagotis brevituba Maxim was extracted with three different solvents: methanol, water, and acetone. In antioxidant assays in vitro, the water extract had the strongest reducing power, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity compared with the methanol and acetone extracts. However, the methanol extract was more potent in the β-carotene/linoleic acid cooxidation assay. In antioxidant assays in vivo, mice that were exposed to 6.0 Gy60Co γ-ray whole-body radiation on day 15 after administration of Lagotis brevituba Maxim decreased their level of malondialdehyde (MDA) in a dose-dependent manner compared with the control group, indicating that Lagotis brevituba Maxim had favorable antioxidant activities in vivo. In addition, a total of 44 compounds were tentatively identified by liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS), including 19 flavonoids, 14 phenols, 8 phenylethanoid glycosides, 2 iridoid glycosides, and 1 carbohydrate. We obtained 25 compounds from plants in the genus Lagotis for the first time. These results suggested that Lagotis brevituba Maxim had potent antioxidant activity and could be explored as a novel natural antioxidant.

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The COL-4A1 polypeptide destroy endothelial cells through the TGF-β/PI3K/AKT pathway

Scientific Reports ◽

10.1038/s41598-021-94801-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ting Li ◽

Zhonghui Ling ◽

Kaipeng Xie ◽

Yixiao Wang ◽

Zhijing Miao ◽

...

Keyword(s):

Endothelial Cells ◽

Rat Model ◽

Vascular Endothelial Cells ◽

Underlying Mechanism ◽

Tube Formation ◽

Cell Counting ◽

Akt Pathway ◽

Control Group

AbstractPreeclampsia (PE) is commonly considered as a placental disorder in pregnancy. Until now, the etiology and pathological mechanism of PE have remained ambiguous. Although PE can lead to a variety of maternal and infant complications, there are still no effective treatments. This study aimed to explore the correlation between the novel polypeptide COL-4A1 and PE, and to identify the underlying mechanism by which this polypeptide may function and to explore new therapeutic targets for PE. A rat model of PE was established and used to verify the function of the polypeptide COL-4A1 in vivo. Additionally, human umbilical vascular endothelial cells (HUVECs) were cultured with or without COL-4A1 and TNF-α (20 ng/ml). Cell Counting Kit-8 (CCK-8), wound-healing, Transwell and tube formation assays were used to evaluate cell proliferation, migration and angiopoiesis. RNA sequencing and mass spectrometry were conducted to explore the underlying downstream mechanism of COL-4A1. In vivo, COL-4A1 increased blood pressure and elevated the risk of fetal growth restriction (FGR) which was induced by lipopolysaccharide (LPS) in the rat model. In vitro, COL-4A1 significantly inhibited the proliferation and migration of HUVECs. After culture with COL-4A1, compared to control group the adhesive ability and level of reactive oxygen species (ROS) were enhanced and tube formation ability was decreased. Furthermore, Western blotting (WB) and pull-down assays were conducted to explore the underlying mechanism by which COL-4A1 functions, and the TGF-β/PI3K/AKT pathway was identified as the potential pathway involved in its effects. In summary, these results revealed that the polypeptide COL-4A1 caused PE-like symptoms in cells and a rat model. Through the TGF-β/PI3K/AKT pathway, COL-4A1 interferes with the pathogenesis of PE. Thus COL-4A1 is expected to become a potential target of PE, providing a basis for exploring the treatment of PE.

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