الجمهورية الجزائرية الديمقراطية الشعبية جامعة فرحات عباس سطيف 1 كلية علوم الطبيعة والحياة University of Ferhat Abbas – Setif 1 Faculty of Natural and Life Sciences وزارة التعليم العالي والبحث العلمي
DERARTMENT OF BIOCHEMISTRY N°…………/SNV/2025
THESIS

Presented by

Benzid Bouthaina

Berkache Ikram

JURY: Chair: BELATTAR Noureddine Pr. UFA Setif 1 Supervisor: BOUTEFAHA Zineddine MCB. UFA Setif 1 Examiner: AMRAOUI Nacer MCA. UFA Setif 1 For the fulfillment of the requirements for the degree of

Master Field: Biological Sciences Special field: Applied Biochemistry Presented in 29/06/2025 Modulation of oxidative stress and hemostasis by methanolic extract of Coriandrum sativum seeds. TOPIC

Laboratory Of Applied Biochemistry 2024-2025

Abstract

This study aimed to evaluate the antioxidant, anticoagulant, and thrombolytic activities of the aqueous-methanolic extract of coriander seeds using in vitro experimental methods. Phytochemical analysis revealed that the extract contains significant levels of phenolic compounds with total phenolic content of 69.87 ± 5.62 µg GAE/mg and total flavonoid content of 26.54 ± 1.45 µg RE/mg. The DPPH assay demonstrated a notable free radical scavenging capacity with an IC₅₀ of 80 ± 5.13 µg/ml. Similarly, the total antioxidant capacity (TAC) test showed an A0,5 of 39.33 ± 4.62 µg/ml, indicating considerable antioxidant potential. Hemolysis inhibition assay revealed a protective effect of the extract on red blood cell membranes, with an IC₅₀ of 0.244 mg/ml, suggesting its role in mitigating oxidative damage. Regarding anticoagulant activity, the extract significantly prolonged clotting times, with PT and aPTT values reaching approximately 57.66 and 68.66 seconds, respectively, at the highest concentration tested (75 mg/ml), indicating interference with both intrinsic and extrinsic coagulation pathways. In thrombolytic assay, fibrinolytic activity increased from 15.82% at 3.125 mg/ml to 35.96% at 50 mg/ml, supporting the extract’s clot-dissolving potential. These findings highlight Coriandrum sativum as a promising natural source for the development of effective therapeutic alternatives targeting oxidative stress, coagulation disorders, and thrombotic conditions, with potential applications in preventive and complementary medicine.

Keywords: Coriandrum sativum, polyphenols, flavonoids, antioxidant activity, anticoagulant activity, thrombolytic activity.

Table of Contents

الملخص	I

Résumé	II

Abstract	III

List of Abbreviations	IV

List of Figures	V

List of tables	VI

Introduction	1

I. Generality about Coriandrum sativum	2

I.1 Common names of plant	2

I.2 Botanical description	2

I.3 Classification of Coriandrum sativum.	3

I.3 Geographical distribution	3

I.4 Chemical composition	3

I.5 Traditional uses of Coriandrum sativum	4

I.6 Biological activities of Coriandrum sativum	4

I.6.1 Antioxidant activity	4

I.6.2 Antimicrobial activity	4

I.6.3 Anti-inflammatory activity	4

I.6.4 Anticancer activity	5

I.6.5 Anti-diabetic activity	5

I.6.6 Neuroprotective activity	5

I.7 Safety of Coriandrum sativum	5

II. Oxidative stress	7

II.1 Definition of free radicals	7

II.2 Sources of free radicals	7

II.3 Major types of reactive oxygen species (ROS)	8

II.4 Body defence mechanisms against free radicals	8

II.5 Antioxidants classification	9

III. Hemostasis	11

III.1 Primary hemostasis	11

III.2 Secondary hemostasis	11

III.2.1 Intrinsic pathway	12

III.2.2 Extrinsic pathway	12

III.2.3 Common pathway	13

III.3 Fibrinolysis	14

III.3.1 Thrombosis	14

III.3.1.1 Venous thrombosis	16

III.3.1.2 Arterial thrombosis	16

III.3.2 Thrombosis treatment	16

III.4 Oxidative stress and coagulation	16

I. Materials and methods	19

I.1Materials	19

I.1.1 Plant material	19

I.1.2 Chemicals	19

I.2 Methods	19

I.2.1Preparation of the hydro-methanolic extract of Coriandrum sativum	19

I.2.2 Determination of total phenolics	19

I.2.3Determination of total flavonoid	20

I.2.4 Antioxidant activity assays	21

I.2.4.1 DPPH free radical scavenging test	21

I.2.4.2 Total antioxidant capacity (TAC) test	21

I.2.4.3 Anti hemolytic activity	22

I.2.5 Evalution of anticoagulant activity	22

I.2.6 Thrombolytic activity	23

II. Statistical analysis	24

III. Results and Discussion	26

III.1 Phenolic compound ratios	26

III.2 Anti oxidant assays	27

III.2.1 DPPH free radical scavenging test	27

III.2.2 Total antioxidant capacity (TAC) test	28

III.2.3 Anti hemolytic activity	29

III.3 Anticoagulant activity	31

III.4 Thrombolytic activity	32

V. Conclusion	35

VI. References	37

Introduction

Introduction

The human body possesses a comprehensive biological defense system against free radicals, comprising enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), in addition to non-enzymatic antioxidants like vitamins C and E, and glutathione. However, any disruption in the balance between free radicals and the body’s natural antioxidants can lead to a condition known as oxidative stress. This imbalance is considered a common underlying factor in the development of various chronic and complex diseases, including cardiovascular disorders, cancer, premature aging, Alzheimer’s disease, and several neurodegenerative conditions (Pizzino et al., 2017).

Interestingly, oxidative stress is also known to influence coagulation pathways, suggesting a possible correlation between antioxidant and anticoagulant strategies (Wang and Zennadi, .2020). On another front, the blood coagulation system plays a vital role in maintaining hemostatic balance and preventing excessive bleeding. Nevertheless, dysregulation of this system can result in a hypercoagulable state, which is associated with severe vascular complications such as stroke, myocardial infarction, and pulmonary embolism. Given the global rise in the prevalence of such conditions, there has been growing interest in identifying safer and more effective therapeutic alternatives to conventional drugs. Although current anticoagulants are effective, their long-term use is often limited by adverse effects, particularly bleeding risks, drug interactions, and the emergence of resistance (Heestermans et al., 2022).

In this context, medicinal plants have gained significant attention as natural sources of bioactive compounds, particularly phenolics and flavonoids, known for their antioxidant, anti-inflammatory, and vasculoprotective properties. These compounds not only inhibit oxidative processes but also modulate inflammatory pathways and protect blood components and vascular tissues from damage. Moreover, their value extends to industrial applications, as they are widely used in the food and pharmaceutical industries as natural antioxidants that enhance product quality and safety (Muscolo et al.,2024).

Among these plants, coriander (Coriandrum sativum), an aromatic and medicinal herb belonging to the Apiaceae family, stands out for its long-standing use in traditional medicine, particularly for treating gastrointestinal disorders, inflammation, and intoxications. Recent literature indicates that coriander contains a range of bioactive compounds with demonstrated antioxidant and anti-inflammatory activities, along with a potential anticoagulant effect (Mahleyuddin et al., 2021). Based on this ethnopharmacological background, the present study aims to evaluate selected biological activities of coriander extract, including its antioxidant, anti-hemolytic, anticoagulant, and thrombolytic properties. Standardized laboratory protocols were adopted to ensure accurate, reproducible, and comparable results, thereby contributing to the expanding body of knowledge on the potential therapeutic applications of this plant.