Complex Probiotics Supplementation Improves Overtraining-Induced Muscle Inflammatory Response and Antioxidant Status Via LPS-TLR4/NF-κb Signalling Pathway in Rats

Abstract Background: Prolonged endurance and intensive exercise has shown detrimental effects on muscle health as well as causes intestinal dysbiosis. Probiotics are known as live microorganisms effective in improving gut microbiota and immunity to warrant health and well-being, which exert global and local biological functions on gut and muscle comprising of anti-oxidative stress, anti-inflammatory and anti-apoptosis, etc,. This research was conducted to investigate whether complex probiotics supplementation could alleviate excessive exercise-induced muscle damage and underlying signaling mechanism in rats.Methods: Thirty-two male SD rats were randomly allocated to four groups: control (SC), exercise (EC), probiotics (SP) and exercise with probiotics (EP) (n = 8 each). In the overtraining model, rats were arranged to an incremental load training on the treadmill 6 days/week for 8 weeks and gavaged with complex-probiotic-preparation (4 × 109 CFU/g) containing maltodextrin or maltodextrin only once a day. Serum and soleus were collected at the end of experiment. Serum creatine kinase (CK) and lactate dehydrogenase (LDH) activities, the levels of lipopolysaccharide (LPS), interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), and muscle total antioxidant capacity (T-AOC), glutathione peroxidase (GPx), catalase (CAT) in soleus were measured. Furthermore, soleus protein expression of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), p-p65/p65, Bax, Bcl-2, caspase-9 and caspase-3 were determined by western blotting assay. Results: The findings indicated that excessive exercise induced a significant increased CK and LDH activities, LPS levels, relative protein expression of TLR4, MyD88, p-p65/p65, inflammatory cytokines and lipid peroxidation (p < 0.05). Simultaneously, the antioxidant elements including T-AOC, GPx, CAT were markedly decreased in the muscle of EC group (p < 0.05). In addition, up-regulated the protein expression of Bax/Bcl-2, cleaved caspase-9/procaspase-9 and cleaved caspase-3/procaspase-3 in muscle (p < 0.05). Administration of complex probiotics attenuated overtraining-induced inflammatory response, oxidative injury and TLR4/nuclear factor-kappa B (NF-kB) signaling factors expression as a result of reversing the most of above altered biochemical parameters (p < 0.05), and there was only tendency downregulation in the expression of apoptosis-regulated proteins (p > 0.05). Conclusions: Administration of complex probiotics could improve capability of the anti-oxidant, anti-inflammatory status induced by excessive exercise in rat skeletal muscle possibly via protecting the gut mucosal barrier, inhibiting the outflow of endotoxin, attenuating the action of TLR4-mediated NF-kB pathway.

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The Effect of Tribulus Terrestris Supplementation on Inflammation, Oxidative Stress and Performance of Recreational Runners: Study Protocol for a Double-Blind Randomized Controlled Trial

10.21203/rs.3.rs-507103/v1 ◽

2021 ◽

Author(s):

Marzieh Nejati ◽

Parvin Dehghan ◽

Mostapha Khani

Keyword(s):

Oxidative Stress ◽

High Intensity ◽

Controlled Trial ◽

Sport Performance ◽

Tribulus Terrestris ◽

Double Blind ◽

Inflammatory Status ◽

Recreational Runners ◽

Anti Inflammatory ◽

Exercise Induced

Abstract Background: High intensity and endurance exercises lead to exercise-induced oxidative stress (EIOS), exercise-induced muscle damage (EIMD), and inflammation, which are the influencing factors on muscle soreness, localized swelling, and sport performance. Therefore, the purpose of this study is to determine the effectiveness of Tribulus terrestris (TT) as an herbal supplement with antioxidant and anti-inflammatory properties on the nutritional, oxidative stress, and anti/inflammatory status, as well as the sport performance of recreational runners.Methods/design: This study is a double-blind, randomized, placebo-controlled trial, which will be conducted among recreational runners of Tabriz stadiums, Iran. Thirty-four recreational runners will be selected, and participants will be assigned randomly to two groups: to receive 500 mg TT supplement or placebo capsules twice daily for two weeks. Both groups will do the high-intensity interval training (HIIT) workouts during the study. Baseline and post-intervention body composition, muscle fatigue, and soreness parameters will be assessed. In addition, assessment of malondialdehyde (MDA), total antioxidant capacity (TAC), superoxide dismutase (SOD), high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), interleukin-10 (IL-10), creatine kinase (CK), lactate dehydrogenase (LDH), insulin-like growth factor-1 (IGF-1) and brain-derived neurotrophic factor (BDNF) will be done during three blood samplings.Discussion: This study will be the first to assess the potential effects of TT in recreational runners. Our results will contribute to the growing body of knowledge regarding TT supplementation on the nutritional, oxidative stress, anti/inflammatory status and sport performance in recreational runners.Trial registration: Iranian Registry of Clinical Trials (www.irct.ir) (ID: IRCT20150205020965N8). Registration date: 13 February 2021.

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LPA protects intestinal epithelial cells from apoptosis by inhibiting the mitochondrial pathway

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00406.2002 ◽

2003 ◽

Vol 284 (5) ◽

pp. G821-G829 ◽

Cited By ~ 54

Author(s):

Wenlin Deng ◽

De-An Wang ◽

Elvira Gosmanova ◽

Leonard R. Johnson ◽

Gabor Tigyi

Keyword(s):

Epithelial Cells ◽

Cytochrome C ◽

Protein Expression ◽

Caspase 3 ◽

Intestinal Epithelial Cells ◽

Intestinal Epithelial ◽

Caspase 9 ◽

Cytochrome C Release ◽

Apoptotic Pathway ◽

Antiapoptotic Activity

We previously showed ( Gastroenterology 123: 206–216, 2002) that lysophosphatidic acid (LPA) protects and rescues rat intestinal epithelial cells (IEC-6) from apoptosis. Here, we provide evidence for the LPA-elicited inhibition of the mitochondrial apoptotic pathway leading to attenuation of caspase-3 activation. Pretreatment of IEC-6 cells with LPA inhibited campothecin-induced caspase-9 and caspase-3 activation and DNA fragmentation. A caspase-9 inhibitor peptide mimicked the LPA-elicited antiapoptotic activity. LPA elicited ERK1/ERK2 and PKB/Akt phosphorylation. The LPA-elicited antiapoptotic activity and inhibition of caspase-9 activity were abrogated by pertussis toxin, PD 98059, wortmannin, and LY 294002. LPA reduced cytochrome c release from mitochondria and prevented activation of caspase-9. LPA prevented translocation of Bax from cytosol to mitochondria and increased the expression of the antiapoptotic Bcl-2 mRNA and protein. LPA had no effect on Bcl-xl, Bad, and Bak mRNA or protein expression. These data indicate that LPA protects IEC-6 cells from camptothecin-induced apoptosis through Gi-coupled inhibition of caspase-3 activation mediated by the attenuation of caspase-9 activation due to diminished cytochrome c release, involving upregulation of Bcl-2 protein expression and prevention of Bax translocation.

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Methamphetamine causes neurotoxicity by promoting polarization of macrophages and inflammatory response

Human & Experimental Toxicology ◽

10.1177/0960327117714039 ◽

2017 ◽

Vol 37 (5) ◽

pp. 486-495 ◽

Cited By ~ 5

Author(s):

X Li ◽

F Wu ◽

L Xue ◽

B Wang ◽

J Li ◽

...

Keyword(s):

Inflammatory Response ◽

Protein Expression ◽

Macrophage Polarization ◽

Enzyme Linked Immunosorbent Assay ◽

Neuronal System ◽

Activated Macrophages ◽

Alternatively Activated Macrophages ◽

Raw264.7 Macrophages ◽

Gene And Protein Expression ◽

Anti Inflammatory

Macrophages, especially their activation state, are closely related to the progression of neurotoxicity. Classically activated macrophages (M1) are proinflammatory effectors, while alternatively activated macrophages (M2) exhibit anti-inflammatory properties. As a powerful addictive psychostimulant drug, coupled with its neurotoxicity, methamphetamine (Meth) abuse may lead to long-lasting abnormalities in the neuronal system. The present study investigated the effect of Meth at subtoxic concentration on macrophage activation state and its underlying toxicity to neuronal cells. PC12 and Murine RAW264.7 cells were coincubated with Meth to test its toxicity. 3-(4,5-Dimethylthiazol)-2,5-diphenyltetrazolium-bromide, enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and Western blot assays were performed to evaluate the toxicity, cytokine secretion, gene, and protein expression. Results showed that cytotoxicity was enhanced on PC12 cells after coculturing with RAW264.7 stimulated with Meth. RAW264.7 macrophages tended to switch to the M1 phenotype, releasing more nitric oxide and proinflammatory cytokines, including tumor necrosis factor α (TNFα), interleukin (IL)-12, and IL-1β, while decreasing the release of anti-inflammatory cytokine IL-10 after treatment with Meth. Meth upregulated the gene expression of IL-6, IL-1β, and TNFα and downregulated the expression of Arg-1, IL-10, and KLF4. Meth could also upregulate the protein expression of IL-1β and TNF α and downregulate the expression of Arg-1 and KLF4. However, the abovementioned effects induced by Meth were abolished by the addition of dopamine receptor D3 antagonist. In conclusion, our study demonstrated that Meth promoted macrophage polarization from M0 to M1 and enhanced inflammatory response, which provided the scientific rationale for the neurotoxicity caused by the chronic use of Meth.

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Anti-Inflammatory and Anti-Oxidant Effect of Dimethyl Fumarate in Cystic Fibrosis Bronchial Epithelial Cells

Cells ◽

10.3390/cells10082132 ◽

2021 ◽

Vol 10 (8) ◽

pp. 2132

Author(s):

Onofrio Laselva ◽

Caterina Allegretta ◽

Sante Di Gioia ◽

Carlo Avolio ◽

Massimo Conese

Keyword(s):

Cystic Fibrosis ◽

Epithelial Cells ◽

Inflammatory Response ◽

Dimethyl Fumarate ◽

Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Inflammatory Status ◽

Anti Oxidant ◽

Anti Inflammatory ◽

Oxidant Effect

Cystic Fibrosis (CF) is caused by mutations on the CF transmembrane conductance regulator (CFTR) gene and is associated with chronic infection and inflammation. Recently, it has been demonstrated that LPS-induced CFTR dysfunction in airway epithelial cells is due to an early oxidative stress. Dimethyl fumarate (DMF) is an approved anti-inflammatory and anti-oxidant drug for auto-immune and inflammatory diseases, but its role in the CF has never been investigated. In this study, we examined the effect of DMF on CF-related cytokines expression, ROS measurements and CFTR channel function. We found that DMF reduced the inflammatory response to LPS stimulation in both CF and non-CF bronchial epithelial cells, both as co-treatment and therapy, and restored LPS-mediated decrease of Trikafta™-mediated CFTR function in CF cells bearing the most common mutation, c.1521_1523delCTT (F508del). DMF also inhibited the inflammatory response induced by IL-1β/H2O2 and IL-1β/TNFα, mimicking the inflammatory status of CF patients. Finally, we also demonstrated that DMF exhibited an anti-oxidant effect on CF cells after different inflammatory stimulations. Since DMF is an approved drug, it could be further investigated as a novel anti-inflammatory molecule to ameliorate lung inflammation in CF and improve the CFTR modulators efficacy.

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JNK pathway promotes hepatocyte apoptosis by inhibiting Bcl-2 and upregulating expressions of Bim, caspase-3 and caspase-9 after cardiopulmonary bypass

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i3.9 ◽

2020 ◽

Vol 19 (3) ◽

pp. 519-524

Author(s):

Haibin Yu ◽

Haojie Zhang ◽

Yan Cheng ◽

Xian’en Fa ◽

Fangtao Zhu ◽

...

Keyword(s):

Cardiopulmonary Bypass ◽

Protein Expression ◽

Caspase 3 ◽

Control Group ◽

Hepatocyte Apoptosis ◽

Caspase 9 ◽

Jnk Pathway ◽

Expression Levels ◽

Over Expression ◽

Mrna Expression Levels

Purpose: To study the effect of Jun N-terminal kinase (JNK) signaling pathway on hepatocyte apoptosis in vivo and in vitro, and to elucidate the mechanism of action. Methods: TdT-mediated dUTP Nick-End Labeling (TUNEL) method was used to determine apoptosis in control and cardiopulmonary bypass (CPB) groups at 0, 3 and 6 hours after rat surgery. The expressions of JNK and p-c-Jun in liver tissues at 0, 3 and 6 h after surgery, and the levels of p-c-Jun, Bcl-2 and Bim following overexpression of JNK, were determined using Western blot assay. Human liver cell line HL-7702 was cultured and transfected with over-expressed JNK plasmid and empty plasmid. Proliferation of HL-7702 cells after JNK over-expression was assessed by Cell Counting Kit-8 (CCK-8), while quantitative real-time polymerase chain reaction (RT-qPCR) was employed to evaluate mRNA expression levels of caspase-3 and caspase-9 mRNA after JNK over-expression. Apoptosis of the cells was determined by flow cytometry (FC) after JNK over-expression. Results: FC results showed that the number of apoptotic hepatocytes increased after JNK overexpression in hepatocytes while TUNEL assay results demonstrated that hepatocyte apoptosis increased in CPB group, when compared to control group; furthermore, the number of apoptotic cells gradually increased within 6 h after surgery. The expressions of JNK and p-c-Jun were higher in CPB group than in control group, and increased gradually in both groups within 6 h after surgery. Overexpression of JNK decreased the proliferation of hepatocytes, and also lowered protein expression levels of p-c-Jun and Bim; on the other hand, the protein expression levels of Bcl-2 fell, while mRNA expression levels of caspase-3 and caspase-9 mRNA increased. Conclusion: JNK pathway promotes hepatocyte apoptosis after cardiopulmonary bypass by inhibiting Bcl-2 pathway and promoting the expressions of Bim caspase-3 and caspase-9. Keywords: Cardiopulmonary bypass, Apoptosis, JNK pathway, Bim, caspase-3 and caspase-9

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Pearl extract protects HaCaT cells from UV radiation-induced apoptosis through mitochondrial pathway regulation

10.21203/rs.2.14302/v1 ◽

2019 ◽

Author(s):

Zhixiong Chen ◽

Jing Wang ◽

Anquan Yang ◽

Lihua Zhang ◽

Yaojia Lu ◽

...

Keyword(s):

Protein Expression ◽

Caspase 3 ◽

Hacat Cells ◽

Oxygen Species ◽

Caspase 9 ◽

Expression Levels ◽

Protein Levels ◽

Bax Protein ◽

Reactive Oxygen Species Content ◽

Induced Apoptosis

Abstract Background: Previous studies demonstrated that pearl extract (PE) promotes wound healing and skin whitening. However, whether PE can inhibit ultraviolet (UV) photodamage in HaCaT cells remains unclear. In this study, an in vitro photoaging cell model was established to observe the effect of PE on UV-induced damage and apoptosis of HaCaT cells. The aim was to provide a reference for future development of natural sunscreen agents. Results: PE concentrations of 0.1 and 1 μg/mL were considered as the most effective and safe concentrations. Compared to the control group, superoxide dismutase and glutathione peroxidase activities in the photoaging group were significantly reduced, while malondialdehyde and reactive oxygen species content, along with tumor necrosis factor-alpha (TNF-a) and interleukin (IL)-10 mRNA and protein levels were markedly increased. In contrast, Bcl-2 protein expression was significantly decreased, while caspase-3, caspase-9, and Bax protein expression levels were significantly increased. Compared to the photoaging group, HaCaT cell proliferation was significantly increased in the PE group. Both PE concentrations significantly increased superoxide dismutase and glutathione peroxidase activities in cells, reduced malondialdehyde and reactive oxygen species content, decreased TNF-a and IL-10 mRNA expression in cells, and reduced TNF-a and IL-10 protein levels in the supernatant. Additionally, Bcl-2 protein expression levels were significantly increased, while caspase-3, caspase-9, and Bax protein expression levels were significantly reduced by PE treatment. Conclusions: PE can inhibit UV-induced apoptosis by inhibiting mitochondria-mediated apoptosis and regulating TNF-a and IL-10 expression.

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Enhancing Cystic Fibrosis Immune Regulation

Frontiers in Pharmacology ◽

10.3389/fphar.2021.573065 ◽

2021 ◽

Vol 12 ◽

Author(s):

Anna M. van Heeckeren ◽

Morgan T. Sutton ◽

David R. Fletcher ◽

Craig A. Hodges ◽

Arnold I. Caplan ◽

...

Keyword(s):

Cystic Fibrosis ◽

Inflammatory Response ◽

Effective Means ◽

Treatment Regimens ◽

Cell Based Therapy ◽

Inflammatory Status ◽

Clinical Consequences ◽

Infection And Inflammation ◽

Anti Inflammatory ◽

Improvement In Survival

In cystic fibrosis (CF), sustained infection and exuberant inflammation results in debilitating and often fatal lung disease. Advancement in CF therapeutics has provided successful treatment regimens for a variety of clinical consequences in CF; however effective means to treat the pulmonary infection and inflammation continues to be problematic. Even with the successful development of small molecule cystic fibrosis transmembrane conductance regulator (CFTR) correctors and potentiators, there is only a modest effect on established infection and inflammation in CF patients. In the pursuit of therapeutics to treat inflammation, the conundrum to address is how to overcome the inflammatory response without jeopardizing the required immunity to manage pathogens and prevent infection. The key therapeutic would have the capacity to dull the inflammatory response, while sustaining the ability to manage infections. Advances in cell-based therapy have opened up the avenue for dynamic and versatile immune interventions that may support this requirement. Cell based therapy has the capacity to augment the patient’s own ability to manage their inflammatory status while at the same time sustaining anti-pathogen immunity. The studies highlighted in this manuscript outline the potential use of cell-based therapy for CF. The data demonstrate that 1) total bone marrow aspirates containing Cftr sufficient hematopoietic and mesenchymal stem cells (hMSCs) provide Cftr deficient mice >50% improvement in survival and improved management of infection and inflammation; 2) myeloid cells can provide sufficient Cftr to provide pre-clinical anti-inflammatory and antimicrobial benefit; 3) hMSCs provide significant improvement in survival and management of infection and inflammation in CF; 4) the combined interaction between macrophages and hMSCs can potentially enhance anti-inflammatory and antimicrobial support through manipulating PPARγ. These data support the development of optimized cell-based therapeutics to enhance CF patient’s own immune repertoire and capacity to maintain the balance between inflammation and pathogen management.

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