Fortilin Interacts with TGF-β1 and Prevents TGF-β Receptor Activation

Fortilin is a 172-amino acid multifunctional protein present in both intra- and extracellular spaces. Although fortilin binds and regulates various cellular proteins, the biological role of extracellular fortilin remains unknown. Here we report that fortilin specifically interacts with TGF-β1 and prevents it from activating the TGF-β1 signaling pathway. In a standard immunoprecipitation-western blot assay, fortilin co-immunoprecipitates TGF-β1 and its isoforms. The modified ELISA assay shows that TGF-β1 remains complexed with fortilin in human serum. Both bio-layer interferometry and surface plasmon resonance (SPR) reveal that fortilin directly bind TGF-β1. The SPR analysis also reveals that fortilin and the TGF-β receptor II (TGFβRII) compete for TGF-β1. Both luciferase and secreted alkaline phosphatase reporter assays show that fortilin prevents TGF-β1 from activating Smad3 binding to Smad-binding element. Fortilin inhibits the phosphorylation of Smad3 in both quantitative western blot assays and ELISA. Finally, fortilin inhibits TGFβ-1-induced differentiation of C3H10T1/2 mesenchymal progenitor cells to smooth muscle cells. A computer-assisted virtual docking reveals that fortilin occupies the pocket of TGF-β1 that is normally occupied by TGFβRII and that TGF-β1 can bind either fortilin or TGFβRII at any given time. These data support the role of extracellular fortilin as a negative regulator of the TGF-β1 signaling pathway

Fortilin potentiates the peroxidase activity of Peroxiredoxin-1 and protects against alcohol-induced liver damage in mice

Fortilin, a pro-survival molecule, inhibits p53-induced apoptosis by binding to the sequence-specific DNA-binding domain of the tumor suppressor protein and preventing it from transcriptionally activating Bax. Intriguingly, fortilin protects cells against ROS-induced cell death, independent of p53. The signaling pathway through which fortilin protects cells against ROS-induced cell death, however, is unknown. Here we report that fortilin physically interacts with the antioxidant enzyme peroxiredoxin-1 (PRX1), protects it from proteasome-mediated degradation, and keeps it enzymatically active by blocking its deactivating phosphorylation by Mst1, a serine/threonine kinase. At the whole animal level, the liver-specific overexpression of fortilin reduced PRX1 phosphorylation in the liver, enhanced PRX1 activity, and protected the transgenic animals against alcohol-induced, ROS-mediated, liver damage. These data suggest the presence of a novel oxidative-stress-handling pathway where the anti-p53 molecule fortilin augments the peroxidase PRX1 by protecting it against degradation and inactivation of the enzyme. Fortilin-PRX1 interaction in the liver could be clinically exploited further to prevent acute alcohol-induced liver damage in humans

การศึกษากลไกด้านภาวะหลอดเลือดแดงแข็งของ morelloflavone จากใบมะพูด

Thesis (Ph.D., Biochemistry)--Prince of Songkla University, 200

Consumption of star fruit juice on pro-inflammatory markers and walking distance in the community dwelling elderly

Purpose: This study aimed to evaluate the effect of star fruit juice supplementation on tumor necrosis factor-alpha (TNF-α), interleukin-23 (IL-23) and interleukin-2 (IL-2), nitric oxide (NO), and 6 min walking distance (6MWD) in a group of elderly individuals. Methods: Twenty-nine individuals (20 males, 9 females) with a mean age of 72.4 ± 8.3 years completed this study. A two-week control period was followed by four weeks of 100 g fresh star fruit juice consumption twice per day after meals. Results: Plasma TNF-α, IL-23, IL-2, NO and the 6MWD were evaluated twice during the control period (weeks 0 and 2) and once after the star fruit juice consumption (week 6). Results: The results showed that all parameters in the blood did not change significantly during the control period. After 4 weeks of star fruit juice consumption, a significant reduction in NO, TNF-α and IL-23 was found; however, there was no change in IL-2. Moreover, the 6MWD increased significantly at week 6, when compared to that at week 0 and 2. Furthermore, the results also showed a significantly positive and negative correlation of NO and TNF-α to the 6MWD, but no correlation of IL-23 and IL-2. Conclusion: This preliminary study concluded that consumption of star fruit juice at 100 g twice daily for one month can significantly depress the pro-inflammation cytokines: TNF-α, IL-23, and NO, while increasing walking distance. Low TNF-α and high NO also present a significant correlation to walking capacity in elderly individuals

Short-Term Pulmonary Rehabilitation for a Female Patient with Chronic Scleroderma under a Single-Case Research Design

Although previously proposed that chronic scleroderma should be cared for clinically and early rehabilitation should be performed in hospital by a chest physical therapist, little evidence is currently available on its benefits. Therefore, this study demonstrated the benefits of short-term pulmonary rehabilitation during hospitalization in a female patient with chronic scleroderma. The aim of rehabilitation was to improve ventilation and gas exchange by using airway clearance, chest mobilization, and breathing-relearning techniques, including strengthening the respiratory system and the muscles of the limbs by using the Breath Max® device and elastic bands. Gross motor function and activities of daily life were regained by balancing, sitting, and standing practices. Data on minimal chest expansion, high dyspnea, high respiratory rate, and low maximal inspiratory mouth pressure were recorded seven days before rehabilitation or at the baseline period. But there was a clinically significant improvement in dyspnea, chest expansion, maximal inspiratory mouth pressure, and respiratory rate, when compared to baseline data, which were recorded by a chest physical therapist during seven days of rehabilitation. Furthermore, physicians decided to stop using a mechanical ventilator, and improvement in functional capacity was noted. Therefore, in the case of chronic and stable scleroderma, short-term rehabilitation during hospitalization for chest physical therapy possibly shows clinical benefits by improving both pulmonary function and physical performance

Effects of L-carnitine supplementation on metabolic utilization of oxygen and lipid profile among trained and untrained humans

Background: The effectiveness of L-carnitine supplementation has been met with conflicting findings when used by sedentary and athletic adults. Objectives: This study aimed to investigate the acute effects of L-carnitine supplementation on aerobic metabolic efficiency and lipid profiles in sedentary and athletic men. Methods: Fifteen sedentary (20.4 ± 1.5 years) and 15 athletic (21.5 ± 2.4 years) men were studied in durations of control, placebo intake and 2 g of L-carnitine supplementation. Lipid profiles, including triglyceride, cholesterol, high-density lipoprotein (HDL) and very-low density lipoprotein (VLDL), were determined before and 40 min after either the placebo or L-carnitine intake. Oxygen consumption (direct VO2), ventilatory threshold (VT), and running time (RT) were recorded after a submaximal treadmill exercise test. Results: Direct VO2 increased significantly at 80% of maximal heart rate after L-carnitine supplementation in both athletic and sedentary men, whereas, a statistical increase in VT and RT occurred only after L-carnitine use in athletes, when compared to the control and placebo subjects. The sedentary group showed no changes in lipid parameters, but triglyceride levels reduced significantly in the athletes after consuming L-carnitine. Conclusions: Acute L-carnitine supplementation possibly affects exercise performance and triglycerides in athletes rather than sedentary men

Ticagrelor induces paraoxonase-1 (PON1) and better protects hypercholesterolemic mice against atherosclerosis compared to clopidogrel.

Ticagrelor (TIC), a P2Y purinoceptor 12 (P2Y12)-receptor antagonist, has been widely used to treat patients with acute coronary syndrome. Although animal studies suggest that TIC protects against atherosclerosis, it remains unknown whether it does so through its potent platelet inhibition or through other pathways. Here, we placed hypercholesterolemic Ldlr-/-Apobec1-/- mice on a high-fat diet and treated them with either 25 mg/kg/day of clopidogrel (CLO) or 180 mg/kg/day of TIC for 16 weeks and evaluated the extent of atherosclerosis. Both treatments equally inhibited platelets as determined by ex vivo platelet aggregation assays. The extent of atherosclerosis, however, was significantly less in the TIC group than in the CLO group. Immunohistochemical staining and ELISA showed that TIC treatment was associated with less macrophage infiltration to the atherosclerotic intima and lower serum levels of CCL4, CXCL10, and TNFα, respectively, than CLO treatment. Treatment with TIC, but not CLO, was associated with higher serum activity and tissue level of paraoxonase-1 (PON1), an anti-atherosclerotic molecule, suggesting that TIC might exert greater anti-atherosclerotic activity, compared with CLO, through its unique ability to induce PON1. Although further studies are needed, TIC may prove to be a viable strategy in the prevention and treatment of chronic stable human atherosclerosis

A preliminary study on the effects of star fruit consumption on antioxidant and lipid status in elderly Thai individuals

Objective: The aims of this preliminary study were to evaluate the antioxidant and lipid status before and after star fruit juice consumption in healthy elderly subjects, and the vitamins in star fruit extracts. Methods: A preliminary designated protocol was performed in 27 elderly individuals with a mean (±SD) age of 69.5±5.3 years, by planning a 2-week control period before 4 weeks of consumption of star fruit twice daily. Oxidative stress parameters such as total antioxidant capacity, glutathione, malondialdehyde, protein hydroperoxide, multivitamins such as l-ascorbic acid (Vit C), retinoic acid (Vit A), and tocopherol (Vit E), and the lipid profile parameters such as cholesterol, triglyceride, high-density lipoprotein-cholesterol (HDL-C) and low-density lipoprotein-cholesterol (LDL-C) were analyzed. Moreover, Vit C, Vit A, and Vit E levels were evaluated in the star fruit extracts during the 4-week period. Results: In the 2-week control period, all parameters showed no statistically significant dif­ference; after 4 weeks of consumption, significant improvement in the antioxidant status was observed with increased total antioxidant capacity and reduced malondialdehyde and protein hydroperoxide levels, as well as significantly increased levels of Vit C and Vit A, when com­pared to the two-time evaluation during the baseline periods. However, glutathione and Vit E showed no statistical difference. In addition, the HDL-C level was higher and the LDL-C level was significantly lower when compared to both baseline periods. But the levels of triglyceride and cholesterol showed no difference. Vit C and Vit A were identified in small quantities in the star fruit extract. Conclusion: This preliminary study suggested that consumption of star fruit juice twice daily for 1 month improved the elderly people’s antioxidant status and vitamins, as well as improved the lipoproteins related to Vit C and Vit A in the star fruit extract