Type 2 diabetes-related proteins derived from an in vitro model of inflamed fat tissue

Exercise improves insulin sensitivity and oxidative capacity in skeletal muscles. However, the effect of exercise on substrate oxidation is less clear in obese and type 2 diabetic subjects than in lean subjects. We investigated glucose and lipid metabolism and gene expression after 48 h with low-frequency electrical pulse stimulation (EPS), as an in vitro model of exercise, in cultured myotubes established from lean nondiabetic subjects and severely obese subjects (BMI ≥ 40 kg/m2) with and without type 2 diabetes. EPS induced an increase in insulin sensitivity but did not improve lipid oxidation in myotubes from severely obese subjects. Thus, EPS-induced increases in insulin sensitivity and lipid oxidation were positively and negatively correlated to BMI of the subjects, respectively. EPS enhanced oxidative capacity of glucose in myotubes from all subjects. Furthermore, EPS reduced mRNA expression of slow fiber-type marker (MYH7) in myotubes from diabetic subjects; however, the protein expression of this marker was not significantly affected by EPS in either of the donor groups. On the contrary, mRNA levels of interleukin-6 (IL-6) and IL-8 were unaffected by EPS in myotubes from diabetic subjects, while IL-6 mRNA expression was increased in myotubes from nondiabetic subjects. EPS-stimulated mRNA expression levels of MYH7, IL-6, and IL-8 correlated negatively with subjects' HbA1c and/or fasting plasma glucose, suggesting an effect linked to the diabetic phenotype. Taken together, these data show that myotubes from different donor groups respond differently to EPS, suggesting that this effect may reflect the in vivo characteristics of the donor groups.

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3D Environment Is Required In Vitro to Demonstrate Altered Bone Metabolism Characteristic for Type 2 Diabetics

International Journal of Molecular Sciences ◽

10.3390/ijms22062925 ◽

2021 ◽

Vol 22 (6) ◽

pp. 2925

Author(s):

Victor Häussling ◽

Romina H Aspera-Werz ◽

Helen Rinderknecht ◽

Fabian Springer ◽

Christian Arnscheidt ◽

...

Keyword(s):

Bone Metabolism ◽

In Vitro Model ◽

Bone Diseases ◽

3D Environment ◽

Type 2 Diabetics ◽

Mineralized Matrix ◽

Vitro Model

A large British study, with almost 3000 patients, identified diabetes as main risk factor for delayed and nonunion fracture healing, the treatment of which causes large costs for the health system. In the past years, much progress has been made to treat common complications in diabetics. However, there is still a lack of advanced strategies to treat diabetic bone diseases. To develop such therapeutic strategies, mechanisms leading to massive bone alterations in diabetics have to be well understood. We herein describe an in vitro model displaying bone metabolism frequently observed in diabetics. The model is based on osteoblastic SaOS-2 cells, which in direct coculture, stimulate THP-1 cells to form osteoclasts. While in conventional 2D cocultures formation of mineralized matrix is decreased under pre-/diabetic conditions, formation of mineralized matrix is increased in 3D cocultures. Furthermore, we demonstrate a matrix stability of the 3D carrier that is decreased under pre-/diabetic conditions, resembling the in vivo situation in type 2 diabetics. In summary, our results show that a 3D environment is required in this in vitro model to mimic alterations in bone metabolism characteristic for pre-/diabetes. The ability to measure both osteoblast and osteoclast function, and their effect on mineralization and stability of the 3D carrier offers the possibility to use this model also for other purposes, e.g., drug screenings.

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3P-168 Development of an in vitro model for adhesive structure : force-dependent assembly of adhesion-related proteins(The 46th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.48.s153_3 ◽

2008 ◽

Vol 48 (supplement) ◽

pp. S153

Author(s):

Daisuke Kiyoshima ◽

Hiroaki Hirata ◽

Hitoshi Tatsumi ◽

Masahiro Sokabe

Keyword(s):

Annual Meeting ◽

In Vitro Model ◽

Biophysical Society ◽

Vitro Model ◽

Related Proteins

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Stress effects on type-1/type-2 cytokine balance: A rapid in vitro model

Journal of Allergy and Clinical Immunology ◽

10.1016/s0091-6749(02)82244-8 ◽

2002 ◽

Vol 109 (1) ◽

pp. S356-S356

Author(s):

Susan Y Ritter ◽

Gailen D Marshall

Keyword(s):

In Vitro Model ◽

Stress Effects ◽

Cytokine Balance ◽

Vitro Model

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Mechanism of Action of Mangifera indica Leaves for Anti-Diabetic Activity

Scientia Pharmaceutica ◽

10.3390/scipharm87020013 ◽

2019 ◽

Vol 87 (2) ◽

pp. 13 ◽

Cited By ~ 2

Author(s):

Dai-Hung Ngo ◽

Dai-Nghiep Ngo ◽

Thi Thanh Nhan Vo ◽

Thanh Sang Vo

Keyword(s):

Sulfonic Acid ◽

In Vitro Model ◽

Amylase Activity ◽

Mangifera Indica ◽

Significant Inhibition ◽

No Production ◽

Vitro Model ◽

Cytotoxicity Effects

Diabetes is a major metabolic disorder whose prevalence is increasing daily. Medicinal plants have played an important role in the prevention and treatment of type 2 diabetes via prophylactic and therapeutic management. In this study, Mangifera Indica leaf (MIL) extract was investigated for its promising anti-diabetic activity via an in vitro model. It was found that MIL extract possessed significant inhibition on alpha-amylase activity up to (51.4 ± 2.7)% at a concentration of 200 µg/mL. Moreover, glucose adsorption capacity of MIL was identified at (2.7 ± 0.19) mM glucose/g extract. Furthermore, the extract caused a significant increase in glucose uptake up to (143 ± 9.3)% in LO-2 liver cells. Notably, MIL extract was effective in scavenging (63.3 ± 2.1)% 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and (71.6 ± 4.3)% 2,2-azinobis-3-ethyl benzothiazoline-6-sulfonic acid (ABTS)+ radicals and inhibiting (66 ± 4.9)% NO production from RAW264.7 cells without any cytotoxicity effects. Accordingly, M. indica leaves are suggested as a promising material for development of hypoglycemic products.

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A novel methylated analogue of L-Mimosine exerts its therapeutic potency through ROS production and ceramide-induced apoptosis in malignant melanoma

Investigational New Drugs ◽

10.1007/s10637-021-01087-5 ◽

2021 ◽

Author(s):

Sotiris Kyriakou ◽

William Cheung ◽

Theodora Mantso ◽

Melina Mitsiogianni ◽

Ioannis Anestopoulos ◽

...

Keyword(s):

Malignant Melanoma ◽

In Vitro Model ◽

Ros Generation ◽

Intrinsic Apoptosis ◽

Membrane Depolarisation ◽

Vitro Model ◽

Induced Apoptosis ◽

Related Proteins ◽

Metabolomic Approach

SummaryMelanoma is an aggressive and highly metastatic type of skin cancer where the design of new therapies is of utmost importance for the clinical management of the disease. Thus, we have aimed to investigate the mode of action by which a novel methylated analogue of L-Mimosine (e.g., L-SK-4) exerts its therapeutic potency in an in vitro model of malignant melanoma. Cytotoxicity was assessed by the Alamar Blue assay, oxidative stress by commercially available kits, ROS generation, caspase 3/7 activation and mitochondrial membrane depolarisation by flow cytometry, expression of apoptosis-related proteins by western immunoblotting and profiling of lipid biosynthesis by a metabolomic approach. Overall, higher levels of ROS, sphingolipids and apoptosis were induced by L-SK-4 suggesting that the compound’s therapeutic potency is mediated through elevated ROS levels which promote the upregulation of sphingolipid (ceramide) biosynthesis thus leading to the activation of both extrinsic and intrinsic apoptosis, in an experimental model of malignant melanoma.

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