اثر تعامل تمرين هوازي با شدت متوسط و اوژنول بر بيان ژن occludin و claudin در بافت قلب رت هاي مسموم شده با كلرپيريفوس

Interaction effect of moderate intensity aerobic training and eugenol on gene expression of claudin and occludin of cardiac tissue of male rats poisoned with chlorpyrifos

زمينه و هدف اتصالات محكم، ساختارهاي پويايي هستند كه عبور يون ها و مولكول ها را در بافت قلب تنظيم مي كنند. هدف از مطالعه ي حاضر، تعيين تاثير تعامل تمرين هوازي با شدت متوسط و اوژنول بر بيان ژن occludin و claudin در بافت قلب رت هاي مسموم شده با كلرپيريفوس بود. روش كار 40 سر رت نر ويستار با ميانگين وزن180 تا220 گرم و سن 8 هفته به طور تصادفي به 5 گروه (8= n) كنترل سالم، كنترل مسموم (كلرپيريفوس)، تمرين هوازي با شدت متوسط + كلرپيريفوس ، اوژنول + كلرپيريفوس و تمرين هوازي با شدت متوسط + اوژنول + كلرپيريفوس تقسيم شدند. تزريق درون صفاقي ml/kg3 كلرپيريفوس و گاواژ mg/kg 250 مكمل اوژنول، به ترتيب به مدت 6 هفته و 4 هفته و هر دو پنج بار در هفته انجام شدند. پروتكل تمرين هوازي شامل 4 هفته تمرين هوازي با شدت متوسط و 5 جلسه در هفته بود. در اولين جلسه ي تمرين، سرعت از m/s 11 و مدت زمان 10 دقيقه دويدن بر روي تردميل شروع شد، به تدريج سرعت و مدت تمرين در طول اجراي پروتكل تمرين افزايش يافت و در جلسه ي آخر سرعت به m/s 20 و مدت زمان 26 دقيقه رسيد. بيان ژن claudinو occludin توسط RT& PCR اندازه گيري شدند. از آزمون هاي تي مستقل، تحليل واريانس دو سويه و آزمون تعقيبي توكي جهت تحليل داده ها استفاده گرديد. يافته ها: نتايج نشان داد، تمرين هوازي با شدت متوسط باعث افزايش معني دار بيان ژن occludin(0/001 = p) و claudin (0/001 = p) در مقايسه با گروه كنترل مسموم شد. اوژنول باعث افزايش معني دار بيان ژن occludin (0/001 =p) شد در حالي كه اثرش بر بيان ژن occludin معني دار نبود (0/391 = p). تعامل تمرين هوازي با شدت متوسط و اوژنول باعث افزايش غير معني دار بيان ژن occludin (0/720 = p) و claudin (0/973 = p). در مقايسه با گروه كنترل مسموم شد. نتيجه گيري: به نظر مي رسد، تمرين هوازي با شدت متوسط و اوژنول از طريق ويژگي آنتي اكسيداني و ضد التهابي خود مي توانند بر بيان ژن هاي درگير در اتصالات محكم بافت قلب موثر باشند.

Insecticides are a very important group of environmental pollutants and their greatest use is in agriculture to protect against diseases and pests, and to improve the quality of crops (1). The use of pesticides and their potential to enter the human diet has raised many concerns for researchers. Organophosphoruses are toxic compounds that people are constantly exposed to in varying concentrations, and its dangers include cancer. Chlorpyrifos is a type of organophosphorus that is widely used as an insecticide in the world. The results of studies have shown that chlorpyrifos has detrimental effects on various organs including the immune system, reproductive system, liver, biochemical and blood parameters. These studies also showed that this toxin increases oxidative stress in various tissues such as liver and kidneys. The role of oxidative stress in cardiovascular disease, Alzheimerchr('39')s, diabetes, and damage to tight junction has been confirmed. Tight junction proteins, as one of the important structural components, are essential for regulating the passage of ions and molecules in the heart. Claudin and occludin are the first densely integral membrane proteins in the blood barrier and have similar topography and function interactively with each other. The permeability of the junction and the function of the epithelial barrier are primarily caused by the claudins, and the types of claudin isoforms are expressed there and determine the specific properties of the tissue. Occludin and claudin are both reduced by oxidative stress. As mentioned, the use of agricultural pesticides such as chlorpyrifos causes oxidative stress in the body. Therefore, it seems necessary to use a method that can protect against the effects of oxidative stress and the side effects of this toxin. Eugenol or 4-ally-2-methoxyphenol is an aromatic molecule found in the oils and essential oils of various plants, including clove and cinnamon. Various studies have shown that natural antioxidants with phenolic structure play an important role in protecting body tissues against free radicals. Based on the evidence presented, eugenol has been shown to have an important protective effect against free radical lipid peroxidation (12) and its anti-inflammatory effect has been confirmed. Today, the use of exercise in increasing cardiovascular fitness and physical fitness, improving oxidative stress and vascular tissue has been approved. The aim of this study was to explain the effect of the interaction of moderate-intensity aerobic training and eugenol on the gene expression of occludin and claudin of the cardiac tissue of rats poisoned with chlorpyrifos Methods 40 male Wistar rats were randomly divided into 5 groups (n = 8): healthy control, poisoned control (chlorpyrifos), moderate intensity aerobic training + chlorpyrifos, eugenol + chlorpyrifos and moderate intensity aerobic training + eugenol + chlorpyrifos. Intraperitoneal injection of 3 ml / kg chlorpyrifos and gavage of 250 ml / kg eugenol supplement were performed five times a week for 6 weeks and 4 weeks, respectively. In order to adapt, rats in the aerobic training group ran for 20 minutes at 9m/min speed for two weeks before starting the main training protocol. The aerobic training protocol consisted of 4 weeks of moderate-intensity aerobic training, 5 sessions per week running on a treadmill. Each training session consisted of 5 minutes of warm-up, 10-26 minutes of moderate-intensity aerobic training (starting at 11 m / s in the first session and reaching 20 m / s in the last session) and 5 minutes of cooling. Gene expression of claudin and occludin were measured by Real time-PCR. Independent t-test, Two-way ANOVA and Tukey post hoc test were used to analysis the data. Results The results showed that the moderate-intensity aerobic training and eugenol caused significant increase in gene expression of occludin (p=0.001) and claudin (p=0.001) compared to Poisoned control group. eugenol caused significant increase in gene expression of occludin (p=0.001) whereas its effect was not significant on occludin gene expression (p=0.391). Interaction effect of moderate-intensity aerobic training and eugenol caused the non significant increase in claudin (p=0.973) and occludin (p=0.720) gene expression in comparison with Poisoned control group. Conclusion Chlorpyrifos toxin increases oxidative stress in various tissues of the body and since oxidative stress affects the expression of claudin and occludin, it significantly reduces the expression of their genes in the blood barrier. However, moderate-intensity aerobic training and eugenol supplementation alone, possibly relying on their antioxidant and anti-inflammatory properties by increasing the activity of antioxidants such as SOD and catalase, reducing lipid peroxidation, activating and increasing ERK1 / 2, and MAPK, decreasing TNF-α, and decreasing beta-amyloid induced the increase the expression of claudin and occludin genes. The results of the present study showed that the interaction of moderate-intensity aerobic training and eugenol did not have a significant effect on the expression of claudin and occludin genes, but what is important is the increase compared to the poisoned control group, and physiologically this change is considered a feat, although the precise mechanism of insignificance cannot be clearly explained. In general, it seems that moderate-intensity aerobic training and eugenol can affect tight junctions.