Oxidized Low-Density Lipoprotein and Upregulated Expression of Osteonectin and Bone Sialoprotein in Vascular Smooth Muscle Cells

Background: Oxidative stress has been associated with the progression of atherosclerosis and activation of genes that lead to increased deposition of proteins in the extracellular matrix. Bone sialoprotein (BSP) and osteonectin are proteins involved in the initiation and progression of vascular calcification. Objective: To investigate the effect of oxidized low-density lipoprotein on osteonectin and BSP expression in human aorta vascular smooth muscle cells (HA/VSMCs). Methods: We treated HA/VSMCs with oxidized low-density lipoprotein (oxLDL) and measured the relative expression of osteonectin and BSP genes using the real-time polymerase chain reaction (PCR) method. We investigated the protein levels produced by each gene using the western blotting technique. Results: oxLDL increased osteonectin and BSP levels (mean SD], 9.1 2.1]-fold and 4.2 0.75]-fold, respectively) after 48 hours. The western blotting results also confirmed the increased levels of osteonectin and BSP. Conclusion: oxLDL may enhance vascular calcification by promoting the expression of osteonectin and BSP

The effect of carvacrol on the interleukin-6 gene expression and cell signaling proteins in prostatic cancer cell line DU-145

زمینه و هدف: اینترلوکین-6 باعث ایجاد خاصیت آنتی آپوپتوزی و مقاومت دارویی در سلول های سرطانی پروستات می شود. هدف از این مطالعه بررسی اثر کارواکرول بر بیان ژن IL-6، پروتئین های سیگنالی آن و توانایی مهاجرت در رده سلول های سرطان پروستات (DU-145) می باشد. روش بررسی: در این مطالعه تجربی آزمایشگاهی، سلول های DU-145 با غلظت های مختلف کارواکرول به مدت 48 ساعت تیمار شدند. سپس، درصد زیست پذیری سلول ها به کمک روش رنگ سنجی تترازولیوم (MTT) اندازه گیری و غلظت مهارکنندگی 50 درصد رشد سلول‌ها (IC50) محاسبه شد. میزان IL-6 در غلظت های مختلف کارواکرول با استفاده از کیت الایزا تعیین شد. بیان ژن IL-6 و پروتئین های سیگنالی آن (pStat3، pErk و pAkt) به ترتیب با استفاده از روش های Real time RT PCR و Western blot تعیین شدند. توانایی مهاجرت و تهاجم سلول های DU-145 با استفاده از تست Invasion بررسی شد. یافته ها: میزان IC50 کارواکرول معادل Mµ2/406 به دست آمد. کارواکرول در غلظت Mµ150 باعث افزایش بیان mRNA مربوط به IL-6 وافزایش سنتز پروتئین آن گردید؛ اما در غلظت های بالاتر از Mµ150 بیان mRNA و سنتز IL-6 به صورت وابسته به دوز کاهش یافت (05/0>P). کارواکرول در غلظت بالاتر از Mµ150 بصورت وابسته به دوز باعث کاهش پروتئین های مسیرهای سیگنالی داخل سلولی pStat3، pErk و pAkt شد؛ همچنین، کارواکرول توانایی تهاجم و زیست پذیری سلول های سرطانی DU-145 پروستات را به طور معنی داری کاهش داد (05/0>P). نتیجه گیری: کارواکرول از طریق کاهش پروتئین های مسیرهای سیگنالی داخل سلولی و بیان ژن IL-6 باعث کاهش رشد، تکثیر و توانایی مهاجرت سلولی و باعث القاء آپوپتوز در سلول های سرطان پروستات (DU-145) می گردد. بنابراین، کاواکرول به عنوان یک ماده درمانی سودمند در درمان سرطان پروستات می تواند مورد توجه قرار گیرد

The Effect of Silymarin on Serum Concentration of Soluble Apoptosis Markers in β-Thalassemia Major Patients Receiving Desferrioxamine

ABSTRACT Background: Despite appropriate chelation therapy with desferrioxamine, iron deposition in visceral organs causes tissue damage in thalassemia major patients. Excess iron can generate reactive oxygen species (ROS) that may lead to cell death and apoptosis. Therefore, antioxidants such as plant flavonoids can be an effective treatment to reduce ROS in thalassemia patients. Aims: In this study, we aimed to investigate the serum levels of apoptosis markers in βthalassemia major patients treated with silymarin and ..

Effect of gallic acid on Alkaline phosphatase gene expression in vascular smooth muscle cells

Background and purpose: Vascular calcification is an important factor in pathogenesis of atherosclerosis. Studies have shown that alkaline phosphatase increases vascular calcification. Here we investigated the effect of gallic acid on alkaline phosphatase gene expression in vascular smooth muscle cells (VSMCs). Materials and methods: In this experimental study humans aorta VSMCs were incubated with beta glycerol phosphate as calcification-inducing media. Then these cells were treated with 160, 180 and 200 µMol concentration of gallic acid for 24h, 48h and 72h. The total RNA was extracted and cDNA was synthesized and then alkaline phosphatase expression was measured by real time PCR. Alkaline phosphatase specific activity was measured by spectrophotometry. Results: Overall, 160, 180 and 200 µMol concentration of gallic acid decreased alkaline phosphatase gene expression in vascular smooth muscle cell by 1.98, 2.03, and 3.16 folds, respectively after 72h compared with the control group. The alkaline phosphatase specific activity also decreased compared to that of the control group. Conclusion: Our results showed that gallic acid decreased the expression and activity of alkaline phosphatase suggesting that this antioxidant compound may attenuate vascular calcification

Effect of Surgical Flap on IL-1β and TGF-β Concentrations in the Gingival Crevicular Fluid of Patients with Moderate to Severe Chronic Periodontitis

Background: Growth factors play a major part in wound healing in many tissues including the periodontium. Transforming growth factor-β1 (TGF-β1) is one of these factors present in the gingival crevicular fluid. In addition, it is considered as one of the most important anti-inflammatory cytokines. Interleukin-1β is a proinflammatory cytokine that presents itself in gingival inflammation and the GCF. Such factors might be of value as prognostic markers of wound healing activity and the therapeutic progress following flap surgery. Objective: The aim of this study was to evaluate the effect of surgical flap on the concentration of IL-1β and TGF-β in the GCF of patients with moderate to severe chronic periodontitis. Methods: The GCF samples were collected, using the Perio-Paper strip at phase 1 (pre-surgery), phase 2 (4th week post surgery) and phase 3 (12th week post surgery) from 20 sites of 10 patients undergoing flap surgery. After the elution, IL-1β and TGF-β concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results: The mean TGF-β and IL-1β concentration decreased from phase 1 to phase 3 (p<0.05). There were no significant statistical correlations between IL-1β and TGF-β1 concentrations in the 3 assessment phases. There was a significant statistical correlation between TGF-β1 concentrations and the Plaque Index (PI) in phase 2 (p<0.05). There was a significant statistical correlation (p<0.05) between IL-1β and TGF-β1 concentration and the probing pocket depth (PPD). Conclusion: The flap surgery has a significant effect on decreasing IL-1β concentration. In the case of TGF-β1, probably the decrease in the concentration after treatment might be due to the removal of the inflammatory stimulants

Study of I405V polymorphism of cholesterol ester transfer protein gene in efficacy of statins on plasma level of high density lipoprotein cholesterol

زمینه و هدف: پروتئین انتقال دهنده کلسترول استر (CETP) در متابولیسم لیپوپروتئین با دانستیه بالا (HDL) و مسیر انتقال معکوس کلسترول نقش اساسی دارد. چند شکلی های ژن CETP مانند I405V (ایزولوسین به والین) که مستقیماً بر HDL کلسترول تاثیر می‌گذارد رونویسی از این ژن را تحت تاثیر قرار می‌دهد. هدف این مطالعه تعیین تاثیر پلی مورفیسم I405V ژن CETP بر سطح HDL کلسترول در پاسخ به درمان با استاتین‌ها می‌باشد. روش بررسی: در این مطالعه توصیفی - تحلیلی از بین بیمارانی که سطح لیپوپروتئین با دانسیته پایین کلسترول (LDL-C) بالاتر از 120 میلی گرم در دسی لیتر تحت درمان، 196 بیمار دریافت کننده لواستاتین و آتوراستاتین انتخاب شدند. در همه بیماران قبل و بعد از درمان پروفایل لیپیدی اندازه‌گیری شد. پلی مورفیسم I405V ژن CETP توسط تکنیک چند شکلی طول قطعه محدود (PCR- RFLP) تعیین گردید. سپس نتایج آزمایشات بیوشیمیایی در پلی مورفیسم‌های مختلف با استفاده از آزمون‌های t زوجی، ANOVA و توکی مورد مطالعه قرار گرفت. یافته‌ها: پس از درمان با لواستاتین در ژنوتیپ VV سطح کلسترول کاهش بیشتر و سطح HDL افزایش بیشتری نسبت به دو ژنوتیپ دیگر نشان داد (05/0

The effect of endurance training intensity on the expression of perlipin-A protein of visceral adipose tissue, serum glucose and insulin levels in STZ-induced diabetic rats

Background and aims: Changes in the expression of lipid droplet adipocyte proteins, such as prelipipin A (PLINA) cause alter lipolysis and insulin resistance. The aim of this study was to compare the three endurance training intensities (low, moderate and high) on the expression of PLINA protein in visceral adipose tissue, serum glucose and insulin levels in male diabetic Wistar rats. Methods: 40 male Wistar rats were assigned to five groups (n=8) including diabetic group with low intensity endurance training, moderate intensity group, high intensity group, diabetic and healthy control groups. After induction of diabetic rats by injection of streptozotocin, endurance training was performed with different intensities for eight weeks, three sessions per week. The relative expression of PLINA protein was measured by western blot technique. One-way variance analysis and Tukey's post hoc test were used to determine the difference between the groups. Results: The results showed that there was a significant difference between PLINA levels in healthy and diabetic control groups with endurance training groups (with low, moderate and high intensity) (P=0.018). These differences were between low intensity training and healthy control groups (P=0.033) and between diabetic and healthy control groups (P=0.020). Serum glucose and insulin levels were significantly different between the diabetic control and endurance training groups (low, moderate and high) (P=0.001). This difference was between high-intensity training group with low intensity training (P=0.046), diabetic control (P=0.001) and healthy control (P=0.011) groups. Conclusion: Moderate and high intensity endurance training can compensate for the loss caused by diabetes in the expression of the PLINA protein and reduces serum levels of insulin and glucose in these mice. It seems that more intensity endurance training leads to more increase in PLINA expression in diabetic rats

Molecular characterization of familial hypercholesterolemia in Iranian patients

Abstract Familial hypercholesterolemia (FH) is an autosomal dominant disorder of lipoprotein metabolism caused mainly by mutations in the low-density lipoprotein receptor (LDLR) and apolipoprotein B 100 (APOB) genes. Until now, the molecular basis of FH has been demonstrated in detail in many populations, but there is still very limited Molecular data concerning FH in Iran. The aim of this study was to characterize the LDLR and APOB gene mutations in an Iranian population. A total of 30 non-related Iranian possible FH subjects were studied. Diagnosis of FH was based on the Dutch Lipid Clinic Network diagnostic criteria. All samples were initially tested for three common APOB gene mutations including R3500Q, R3500 W and R3531C using PCR-RFLP assay. Subsequently, promoter and coding region of the LDLR gene was screened by PCRSSCP analysis and positive results were confirmed by DNA sequencing. Four previously reported polymorphisms 1413G [A, 1725C [T, 1773T [C and 2140 ? 5G[A were found in *17% (5/30) of population studied. Moreover, no variation was found in APOB gene. Our data indicated that LDLR and APOB gene mutations have not contribution to possible FH in Iranian population studied here. However, we examined three common APOB mutations and LDLR in only 30 patients, and to determine the role of these genes in developing FH in Iran, more FH samples and populations needed to be investigated for the mutations of the related gene

Molecular pathology of 6 novel GJB2 allelic variants detected in familial and sporadic Iranian non syndromic hearing loss cases

Background: Mutations of GJB2 gene encoding connexion 26 are the most common cause of hearing loss in many populations. A very wide spectrum of GJB2 gene mutations associated with hearing loss have been detected but pathogenic role has been tested only for a part of them. In this study, we have provided genetic evidence on the pathogenicity of our previously reported novel GJB2 allelic variants. Methods: The pathogenic role of GJB2 allelic variants were assessed using co segregation of each allelic variant with hearing loss in family members, absence of the allelic variants in control populations, coexistence with a second GJB2 mutation, nature of the amino acid substitution and evolutionary conservation of the appropriate amino acid. Results: The GJB2 allelic variants including 363delC, 327delGGinsA, H16R and G200R have been co segregated with autosomal recessive non syndromic hearing loss in five families and are not found in control subjects. The G130V and K102Q were found in heterozygous state in two deaf individuals. G130V results in an exchange a residue highly conserved among all the connexins but was found with a rate of 1% in control subjects and K102Q results in an exchange a residue not conserved among all the connexins and not identified in control subjects. Conclusion: We conclude that, 363delC, 327delGGinsA, H16R and G200R may be pathogenic. However, the pathogenicity and inheritance of K102Q and G130V can not be assessed clearly and remains to be identified

Oxidized Low-Density Lipoprotein Increases Bone Sialoprotein Expression in Vascular Smooth Muscle Cells Via Runt-Related Transcription Factor 2

Background: Vascular calcification is a pivotal stage in atherosclerosis. During vascular calcification, vascular smooth muscle cells (VSMCs) synthesize many osteogenic factors such as bone sialoprotein (BSP). Oxidative stress plays a critical role in progression of atherosclerosis and also increases extracellular matrix proteins expression. BSP overexpression has been observed during vascular calcification by oxidative stress. However, the regulatory mechanism of oxidized low-density lipoprotein (oxLDL)-mediated vascular calcification has not yet been fully defined. In this study, we aimed to investigate whether runt-related transcription factor 2 (Runx2) affects the oxLDL-induced BSP expression or not. Methods: In this experimental study, we cultured VSMCs in F12K media and then treated them with oxLDL. The expression of Runx2 and BSP genes was determined by real-time polymerase chain reaction method. Protein level of each gene was investigated by Western blotting technique. To determine whether Runx2 regulates BSP gene expression at VSMCs induced by oxLDL, we suppressed Runx2 mRNA using siRNA. Transfected cells then were treated with oxLDL and expression of Runx2 and BSP genes was determined again. Results: oxLDL increased Runx2 and BSP expression (4.8 +/- 0.47-fold and 4.91 +/- 0.56-fold, respectively) after 48 hours. Western blotting method confirmed the increased levels of Runx2 and BSP proteins after 48 hours. Runx2 overexpression alone induced BSP expression, whereas knockdown of Runx2 with small interfering siRNA blocked oxLDL-induced BSP expression. Conclusions: Our results showed that oxLDL-induced BSP expression was dependent on Runx2 expression, suggesting that Runx2 is required for oxLDL-induced BSP expression