Volume 30, Issue 2 (4-2023)                   RJMS 2023, 30(2): 25-39 | Back to browse issues page

Ethics code: IR.BASU.REC.1398,024

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Shahabi Baher I, Mirazi N, Izadi Z, Nourian A, Safari S. Study of Antioxidant and Anti-inflammatory Effects of Calendula officinalis L. Hydrethanolic Extract on Bone Marrow Tissue in Wistar Rats Treated with Cyclophosphamide. RJMS 2023; 30 (2) :25-39
URL: http://rjms.iums.ac.ir/article-1-7431-en.html
Professor, Department of Biology, Faculty of Basic Sciences, Bu-Ali Sina University, Hamedan, Iran , mirazi205@gmail.com
Abstract:   (1103 Views)

Background & Aims: Some drugs used in chemotherapy have destructive effects on hematopoietic tissue.Cyclophosphamide is a commonly used anti-cancer drug which used for some of malignant cancers treatment and causes toxicity by its reactive metabolites such as acrolein and phosphoramide mustard. This drug has oxidant, inflammatory and subversion effects on blood hemopoetic bone marrow tissue. Medicinal plants with antioxidant and anti-inflammatory effects have protective effects on bone marrow tissue. The aim of this experimental study is to investigate  antioxidant and anti-infammatory effects of  Calendula officinalis L. hydroethanolic extract (CHE) on bone marrow in Wistar rats treated with cyclophosphamide.
Methods: In this experimental study 35 male Wistar rats (220-250 g) were divided in 5 groups (n=7): control (Normal saline, 0.5 ml), Cyclophosphamide (CP, 15mg/kg), positive control (CHE, 600 mg/kg), treatment 1 (CP, 15mg/kg + CHE, 300 mg/kg) and treatment 2 (CP, 15mg/kg + CHE, 600 mg/kg). All injections were done interaperituneally and the experiments were done for 15 days. At the end of experiments all animals were anesthetized ( ketamine hydrochloride, 50mg/kg + xylasine hydrochloride, 10mg/kg). After splitting the chest with surgical scissor, blood samples were collected directly from the right ventricle of the heart. The collected blood samples were centrifuged at 4000 rpm for 5 minutes and their serums were separated. Serums prepared send to the laboratory for  measuring serum parameters such as: Nitric oxide(NO), Glutamine (GSH), Malondialdehyde (MDA), Glutathione peroxidase (GPx), Tumor Necrosis Factor- alfa(TNF-α) and Inetrlukin-6β (IN-6β). In addition, rat sternum's tissue samples were isolated and fixed with 10% Formaldehyde. Bone marrow tissue samples were studied in the histology laboratory after various steps and preparation of microscopic sections. Data were analyzed using one-way analysis of variance and Tukey's test. The significant difference of the data were considered with p <0.05.
Results: The results of this study showed that cyclophosphamide caused a significant increase in serum TNF-α levels in the group receiving cyclophosphamide compared to the control group (p<0.001). While in the positive control group and treatments 1 and 2 groups, there was no significant difference compared to the control group. Also, there was no significant difference between treatment 1 and treatment 2 groups. Cyclophosphamide significantly increased serum IL-6β levels in the CP group compared to the control group (p<0.001). However, serum levels of IL-6β in the groups treated with CHE were significantly reduced compared to the CP group. There was significant difference of serum levels of IL-6β between treatment 1 and treatment 2 groups (p<0. 01). The use of cyclophosphamide showed that the NO serum level in the CP group was significantly higher than the control group. The groups treated with CHE had a significant decrease in NO serum level compared to the CP group (p<0.001). There was significant difference of serum levels of NO serum level between treatment 1 and treatment 2 groups (p<0. 001). In addition, MDA serum levels in the CP group were significantly increased compared to the control group. There was significant difference of serum levels of MDA serum levels between treatment 1 and treatment 2 groups (p<0. 001). There was significant differences of MDA serum levels in CP group, positive group  and treatment groups 1&2 (p<0. 001). Another result of this study was that cyclophosphamide significantly reduced serum levels of GSH and GPx in the CP group compared with control group (p<0. 001). While serum levels of these parameters in positive control groups and treatments 1 and 2 showed a significant increase compared to the CP group. There were no significant differences of GPx and GSH serum levels between treatment groups 1&2. Histological results in this study showed that CP reduces cell density, loss of cell cohesion and also infiltration of lymphocytes in bone marrow tissue samples in CP group. These results also showed that in treatment group 1 cell density is relatively higher than CP group. In treatment group 2, cell density increased and a significant decrease in inflammatory cells were observed. No significant changes were observed in the positive control group compared to the control group.
Conclusion: The results of this study showed that cyclophosphamide has destructive effects on various tissues of the body, including hematopoietic tissue of bone marrow in test rats. The effects of oxidative stress and cell death of cyclophosphamide have been previously reported. The use of drugs used in the process of chemotherapy is very common and inevitable. Cyclophosphamide, which is used in pediatric oncology as part of standard treatment for leukemia, solid tumors and brain tumors, is one of these drugs. Cyclophosphamide, as a cytotoxic drug, causes inflammation and increases inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α),  IL-1β and IL-6β. Cyclophosphamide has been identified as one of the most important tissue toxicity agents due to its reactive metabolites such as acrolein and phosphoramide mustard and it has been shown that this anticancer drug increases inflammatory serum factors such as TNF-α and nitric oxide (NO) and decreases regenerative glutathione (GSH). In various studies, the antioxidant effects of CHE extract have been proven. Many chemical compounds have been identified in CHE, including carotenoids, flavonoids, terpenoids, coumarins, quinones, amino acids, lipids, and carbohydrates. These compounds can be used for various medicinal and therapeutic purposes as an antioxidant, anti-inflammatory, antibacterial, antifungal, anti-cancer, anti-HIV, for wound healing and more. The main chemical constituents of CHE include steroids, terpenoids, free and esterified terterpene alcohols, phenolic acids, flavonoids (quercetin, rutin, narcissin, isoramentin, campferrol) and other compounds. Research has shown that coumarins have very beneficial effects on cell proliferation and preventing apoptosis. Large amounts of coumarin have been mentioned in the extract of Calendula officinalis's extract. Coumarins have been used to treat prostate cancer, renal cell carcinoma and leukemia. In addition, coumarins play an important role in preventing side effects from radiotherapy. The antioxidant activity of CHE was shown in a study performed on polymorphonuclear leukocytes. Flavonoids and phenolic compounds with iron-binding region have iron chelating properties. By flaking iron, flavonoids both absorb more iron from the intestines and prevent it from being excreted by the kidneys. By doing this, flavonoids store iron in tissues, especially in hematopoietic tissues. Calendula officinalis's flowers contain large amounts of antioxidant compounds (flavonoids and polyphenols), which indicates the antioxidant properties of this plant. The presence of propylene glycol in CHE has been effective in preventing the formation of oxidizing free radicals such as ROS and RNS in isolated leukocytes in the respiratory tract. The protective effects of CHE on inflammatory parameters and serum antioxidants seem to be related to its antioxidant and anti-inflammatory effects. Therefore, it is thought that CHE extract, due to its antioxidant and anti-inflammatory agents, is able to prevent the destructive effects of cyclophosphamide on hematopoietic tissue.

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Type of Study: Research | Subject: Physiology

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