Acyl ghrelin, desacyl ghrelin and their ratio affect hepatic steatosis via PPARγ signaling pathway.

BACKGROUND AND STUDY AIMS: Ghrelin is an appetite hormone-containing 28-amino acid and has 4 different forms in the body. Ghrelin forms have different physiological functions in the body. This study aims to analyze the effect of acyl and desacyl ghrelin hormone on hepatic steatosis and biochemical findings in 36 male Wistar rats. MATERIALS AND METHODS: Rats were split into 6 equal groups, consisting of control, acyl ghrelin, desacyl ghrelin, acyl/desacyl 3:1, acyl/desacyl 1:1, and acyl/desacyl 1:3 groups, and administered placebo or 200 ng/kg hormone subcutaneous twice a day for 14 days. Oral Glucose Tolerance Test (OGTT) was performed on Day 15, Insulin Tolerance Test (ITT) on Day 16, and scarification procedure on Day 17. Certain biochemical data and liver diacylglycerol (DAG), glycogen, protein kinase C and PPAR-γ levels were detected in the blood. Histological analyses were also conducted on the liver tissues. RESULTS: The highest plasma total cholesterol and VLDL-K levels were found in the acyl/desacyl 1:3 group, and lower insulin, and HOMA-IR levels were found in groups where acyl and desacyl were administered together (p < 0.05). PPAR-γ gene expression level increased in acyl ghrelin and acyl/desacyl 1:3 groups compared to the control group. Protein kinase C gene expression was highest in the acyl/desacyl 1:3 group. The most severe degenerative findings compliant with steatosis in the liver were observed in the acyl ghrelin group (p < 0.05). CONCLUSION: It was determined that administering rats acyl alone and acyl/desacyl by 1:3 caused the highest PPAR-γ gene expression, serum total cholesterol, HDL-K, and VLDL-K levels in the body. Besides, it is shown that desacyl ghrelin effectively regulates the blood glucose level when administered alone.

Resveratrol and quercetin protect from Benzo(a)pyrene-induced autophagy in retinal pigment epithelial cells.

PURPOSE: This study aims to investigate the role of Resveratrol (RES) and quercetin (QR) treatments against Benzo(a)pyrene (B(a)p)-induced autophagy in retinal pigment epithelial cells. METHODS: The IC50 doses of B(a)p, RES and QR in retinal pigment epithelial cells were determined by MTT assay and the relevant agents were administered singly or in combinations to ARPE-19 cells for 24 h. Occurrence of autophagy in the cells was verified by detection of autophagosomes using fluorescence microscope. Also, the mRNA expression levels of LC3 and Beclin 1 genes were analyzed by RT-PCR to collect further data on autophagy. Caspase-3 and IL-1ß levels in lysed cells were analyzed by ELISA. RESULTS: Autophagosomes were detected in B(a)p-treated ARPE-19 cell lines, as well as a 1.787-fold increase in LC3 mRNA expression levels. No autophagosome occurred in RES and QR treatments, and a significant decrease in their percentage amounts were observed in B(a)p + RES and B(a)p + QR. The mRNA expression levels of LC3 and Beclin 1 also supported these findings. B(a)p had no effect on Caspase-3 levels in ARPE-19 cells, but combined with RES and QR, it increased Caspase-3 levels significantly.IL-1ß levels were higher in B(a)p, B(a)p + QR, B(a)p + RES, RES and QR than control group. This rise in IL-1ß levels was correlated with suppression of mRNA expression levels of Beclin 1. CONCLUSION: B(a)p exposure caused autophagy in ARPE-19 cells, but did not induce apoptosis. RES and QR treatments prevented B(a)p-induced autophagy. Therefore, RES and QR treatments showed protective effect against potential degenerative diseases caused by chronic exposure to B(a)p.

Daidzein nanosuspension in combination with cisplatin to enhance therapeutic efficacy against A549 non-small lung cancer cells: an in vitro evaluation.

Lung cancer is the most common cause of cancer-related mortality, chemo-resistance, and toxicity limit treatment. The focus is on innovative combined phytotherapy to improve treatment outcomes. Our aim was to investigate the potential effects of daidzein nanosuspension (DZ-NS) and its combination with cisplatin (CIS) on A549 non-small lung cancer cells. Cytotoxicity was investigated using MTT and Chou-Talalay methods. Oxidative, apoptotic, and inflammatory markers were analyzed by ELISA and qRT-PCR. The IC50 value for DZ-NS was 25.23 µM for 24 h and was lower than pure DZ (IC50 = 835 µM for pure DZ). DZ-NS (at IC50x2 and IC50 values) showed synergistic cytotoxicity with CIS. The cells treated with DZ-NS had low TOS and OSI levels. However, DZ-NS failed to regulate Cas3 and TGF-ß1 activation in A549 cells. MMP-9 gene expression was significantly suppressed in DZ-NS-treated cells, especially in combination therapy. DZ represents a potential combination option for the treatment of lung cancer, and its poor toxicokinetic properties limit its clinical use. To overcome these limitations, the effects of the nanosuspension formulation were tested. DZ-NS showed a cytotoxic effect on A549 cells and optimized the therapeutic effect of CIS. This in vitro synergistic effect was mediated by suppression of MMP-9 and not by oxidative stress or Cas3-activated apoptosis. This study provides the basis for an in vivo and clinical trial of DZ-NS with concurrent chemotherapy.

An investigation into the cellular-level adverse effects of tourniquet use on the infrapatellar fat pad in primary total knee arthroplasty: A prospective randomized study.

OBJECTIVE: This study aimed to examine the cellular-level adverse effects of tourniquet use on the infrapatellar fat pad (IPFP) in patients undergoing primary total knee arthroplasty (TKA). METHODS: Infrapatellar fat pad samples were collected in a prospective, randomized design to compare 2 groups of primary TKA patients with a tourniquet (T) and without a tourniquet (NT). The study included 80 knees of 58 patients with a mean age of 65.91 ± 9.04 years. The authors collected 3 samples from the T group (after exposure to the fat pad "t1," just before deflating the tourniquet "t2," just before fascia closure "t3") and 2 samples from the NT group (t1 and t3) for each patient. BAX, Bcl-2, and HIF-1α staining showed the extent of cellular hypoxia and apoptosis in IPFP cells, whereas the oxidative stress index (OSI) was determined using a biochemical method. The Knee Injury and Osteoarthritis Outcome Score (KOOS), Knee Society Score (KSS), and Kujala score were used as clinical outcome measures. RESULTS: The mean HIF-1α, BAX/Bcl-2, and OSI scores across all time points were significantly higher in the T group than in the NT group (p<0.001) (d=1.16, 2.9, and 0.9, respectively). The mean BAX/Bcl-2 (P=.030) and HIF-1α (P < .001) scores significantly peaked at t2 in the T group (d=-1.2 and -3.9, respectively). The OSI had higher levels at t1 (P=.011) and t3 (P=.073) (d=0.2 and 0.1, respectively) than at t2 in the T group. The third-month postoperative follow-up revealed that the mean KOOS, KSS, and Kujala score improved significantly compared to the baseline preoperative values (P < .001); however, there was no difference between the T and NT groups regarding the maximum and total knee range of motion or clinical outcome scores. CONCLUSION: Evidence from this study has shown that tourniquet use during primary TKA may be associated with significantly increased cellular hypoxia, oxidative stress, and apoptosis in the IPFP. LEVEL OF EVIDENCE: Level I, Therapeutic study.

Low-dose rosmarinic acid and thymoquinone accelerate wound healing in retinal pigment epithelial cells.

PURPOSE: Thymoquinone (TQ) and rosmarinic acid (RA) are two biologically active compounds found in plants and that possess remarkable anti-oxidant and anti-inflammatory properties. The present study aimed to investigate the potential protective effects of RA and TQ, which have known anti-inflammatory and anti-oxidant effects, on retinal damage by establishing a wound healing model for retinal pigment epithelial cells (ARPE-19). METHOD: To this end, IC50 doses of RA and TQ in ARPE-19 cells were calculated by MTT assay. Both agents were administered at IC50, IC50/2 and IC50/4 doses for wound healing assay, and wound closure percentages were analyzed. Since the best wound healing was found at IC50/4 dose (low dose) for both agents, other biochemical and molecular analyses were planned to be performed using these doses. Following low dose RA and TQ treatments, the cells were lysed and TGF-ß1 and MMP-9 levels were analyzed by ELISA technique from the cell lysates obtained. In addition, the mRNA expression levels of TLR3, IFN-γ and VEGF were calculated by RT-PCR technique. RESULTS: Low dose of RA and TQ dramatically increased wound healing. RA may have achieved this by increasing levels of MMP-9 and TLR-3. In contrast, the mRNA expression level of VEGF remained unchanged. TQ accelerated wound healing by increasing both the protein levels of TGF-ß1 and MMP-9. Furthermore, low dose of TQ decreased both TLR3 and IFN-γ mRNA expression levels. CONCLUSION: Low doses of RA and TQ were clearly demonstrated to have protective properties against possible damage to retinal pigment epithelial cells.

Effects of quercetin on hepatic fibroblast growth factor-21 (FGF-21) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) levels in rats fed with high fructose.

BACKGROUND: Available studies show that quercetin reduces Metabolic Syndrome (MetS) and its complications, increases insulin sensitivity and improves glucose levels. It has been reported that the increase in hepatic gene expressions of fibroblast growth factor-21 (FGF-21), an important metabolic regulator of insulin sensitivity, glucose and energy homeostasis, and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), which plays a central role in the regulation of cellular energy metabolism, eliminate the negative effects of fructose in fructose-fed rats. The main purpose of our study is to examine the effects of quercetin on hepatic FGF-21 and PGC-1α expressions and levels, as well as its protective and therapeutic role on MetS components in rats fed with fructose. METHODS AND RESULTS: In our study, 24 Sprague Dawley male rats were divided into 4 groups: control, fructose, quercetin, fructose+quercetin (n = 6). During the 10-week experiment, quercetin was administered at a daily dose of 15 mg/kg body weight and fructose at a rate of 20%. Blood pressure and weights of all groups were measured and recorded. At the end of week 10, blood and liver tissue samples were taken. Serum insulin, glucose and triglyceride, total, HDL and VLDL cholesterol levels were determined from the samples. Insulin resistance was calculated using the HOMA-IR formula. Hepatic PGC-1α and FGF-21 protein levels and their mRNA expressions were determined. Criteria for metabolic syndrome were successfully established with fructose. It was observed that the administration of quercetin alone and in combination with fructose exerted positive effects and improved MetS criteria. It was determined that the administration of quercetin increased hepatic FGF-21 and PGC-1α protein levels and Messenger RNA (mRNA) expressions of them, which were decreased by fructose application. CONCLUSIONS: The results of our study showed that 10-week administration of quercetin at 15 mg/kg exerted beneficial effects on lipid and carbohydrate metabolism in the fructose-mediated MetS model; therefore, quercetin may have great potential in the prevention and treatment of metabolic disorders.

The effect of acetylsalicylic acid on vasopressin, serum insulin levels, insulin resistance, and biochemical parameters in rats induced with experimental diabetes type 2.

BACKGROUND/AIM: Acetylsalicylic acid (ASA) treatment in diabetic patients is very important owing to the increasing hyperactivity of thrombocytes and atherosclerosis. In several investigations, it was reported that diabetes caused increased coronary artery disease, cerebrovascular disease, and death. In this study, we aimed to investigate the effect of ASA on osmoregulation, glycemic control, and some biochemical parameters in rats induced with experimental diabetes type 2. MATERIALS AND METHODS: Twenty-four rats were randomly divided in four groups: control (I), ASA control (II), diabetic (III), and ASA diabetic (IV). Diabetes was induced by streptozotocin treatment (30 mg/kg, twice, intraperitoneal injection) in obese rats. ASA (150 mg/kg body weight, orally) was administered for 5 weeks in the ASA control and ASA diabetic groups. Serum electrolytes, creatinine, albumin, and total protein levels were analyzed with an autoanalyzer. Arginine vasopressin (AVP) and insulin were analyzed by ELISA techniques. RESULTS: At the end of the study ASA treatments had decreased the fasting blood glucose levels but had interestingly increased the serum AVP levels in diabetics rats. CONCLUSION: AVP levels were increased 2-fold by ASA treatment in diabetic rats. For the first time in this study, the hypoglycemic effect of ASA was attributed to an increase in blood volume by AVP levels. This explanation may be a new approach to the literature on this topic.

Assessment of inflammatory biomarkers and oxidative stress in pulmonary thromboembolism: follow-up results.

Instability in circulation, hypoperfusion, hypoxia, and ischemia in pulmonary thromboembolism (PTE) may occur as a result of failure in pulmonary circulation. All these conditions cause inflammation and oxidative stress. We aimed to investigate inflammatory markers, asymmetric dimethylarginine (ADMA) levels, and the oxidant-antioxidant balance in patients with PTE. This study was conducted as a prospective case-control study. Thirty-eight patients with PTE enrolled to the study. Age- and gender-matched 38 healthy subjects without risk factors for pulmonary embolism were selected as control group. Venous blood samples were obtained from the PTE patients during the initial diagnosis and at the first month of treatment and from the control subjects. Interleukine-6 (IL-6), tumor necrosis factor alpha (TNF-α), total antioxidant status (TAS), total oxidant status (TOS), and ADMA levels were measured for all the samples. The results of patients and healthy subjects were compared. The mean age of the control group was 51.81 ± 15.18 years, and the mean age of the patients was 52.90 ± 18.22 years (p = 0.770). Deep venous thrombosis was present in 68 % of the patients. While we found significant differences between the patient and control groups in terms of IL-6, TAS, TNF-α, ADMA and oxidative stress index (OSI) values (p = 0.001, p = 0.011, p = 0.038, p = 0.028, and p = 0.024, respectively), the TOS value was not different between the groups (p = 0.080). The ADMA, TNF-α, TAS, TOS, and OSI values of the patients during the initial diagnosis and at the first month of treatment were not different (p > 0.05). The results of this study indicate an increased inflammation, endothelial damage, and oxidative stress in PTE. No difference at the first month of therapy suggests ongoing processes. We consider that these markers may be useful in the diagnosis and follow up of PTE.