College of science Department of Botany and Microbiology Application of Spirulina to enhance Liver’s Functions: Effects and Safety Prepared by: Wahida Haza’a Al qahtani Supervised by: Prof. Ali Abdullah Al-Homaidan Prof. Ammar Cherkess Alrikabi Submitted in partial fulfillment of the requirement for the Doctor of Philosophy degree in the Department of Botany and Microbiology at College of science, King Saud University 1436-1437 II I بسمميحرلا نمحرلا هللا قال هللا تعالى في كتابه الكريم: ]ﭤ ﭥ ﭦ ﭧ ﭨ ﭩ ﭪ ﭫ ﭬ ﭭ ﭮ ﭯ ﭰ ﭱ ﭲ ﭳ ﭴ ﭵ ﭶ ﭷ[ صذق هللا العظيم آل عمران : 18 II إهذاء إليكم اخوتي أحبتً .... إلى قلوب كانت صغٌرة، أخذت اسقٌها من دمع عٌنً حرقة وشوقا. ومن دم قلبً لهفة وحبا، حتى كبرت وصار قلبً ٌرقص فرحا حٌن لقٌاها. إلى قلوب ماتزال صغٌرة، أسقٌها من دمً وروحً وتسقٌنً من ثغرها البريء باسمة ومتفائلة وحانٌة. إليكم أوﻻدي صوت فرحتً بل سر قوتً.... إليك أبي " ٌا حبٌبً أنت" مت جسدا وعشت روحا ذائبة فً روحً.... إليك أمي " ٌا حبٌبتً أنت" نهر العطاء الذي ﻻ ٌنتهً. أنت ما أنت !!! "أنت أم ﻻ كاﻷمهات ".... إليك نفسي... من الحٌاة تألمت وتعلمت ... إليك وحيــــــــــــــــــــــــده.... III Acknowledgement This project was very difficult and it was hard to imagine that it will be finished successfully. There are a lot of people who supported and encouraged me in every single moment of this studying period. I was not familiar with some of the laboratory techniques which were not a part of any previous training. But there were many well-wishers who supported me to learn those techniques. I owe sincere thanks to all of them for extending their sincere support and help and for teaching me the techniques during the course of my project study. This project was supported by a grant from the "research center of the female scientific and medical colleges, "Deanship of Research, King Saud University and King Abdul-Aziz City of Science and technology (KACST). Indeed thanks to them for helping and supporting me through this study. I would like to thank Prof. Dr. Ali Al-Homaidan for supervising, encouraging and supporting me in this study. Dr. Mohammad Al Turky and DR. Abdallah Al Aseery from King Faisal Specialist Hospital and Research Center; for their kind help. Mrs. Eidah Alenzi for her patience in teaching me, and sharing her statistical knowledge. Finally, I would like to thank Dr. Taisier Almunaizel for training me to handle the experimental animals during the course of my study. I am also thankful for the kind treatment I received like a daughter. IV Table of Contents إهذاء ........................................................................................................................................ II Acknowledgement .............................................................................................................. III Table of Contents ............................................................................................................... IV List of Tables ..................................................................................................................... VII List of Figures .................................................................................................................. VIII List of Abbreviations ......................................................................................................... IX Abstract ............................................................................................................................... XI 1. Chapter one: Introduction: .............................................................................................. 1 2. Chapter two: Literature review: ..................................................................................... 8 2.1. Overview of Spirulina ................................................................................................ 8 2.2. Biology of Spirulina .................................................................................................. 10 2.3. Classification of Spirulina ........................................................................................ 10 2.4. History of Spirulina .................................................................................................. 12 2.6. Cultivation of Spirulina: .......................................................................................... 13 2.7. Spirulina Photosynthetic pigments content:........................................................... 15 2.8. Physiology of Liver: ................................................................................................. 15 2.9. Anatomy of the Liver: .............................................................................................. 16 2.10. Antioxidant: ............................................................................................................ 16 2.10.1. Classification of antioxidant: ............................................................................. 17 2.10.1. Glutathione: ................................................................................................................... 17 2.10.2. Melatonin: ..................................................................................................................... 18 2.11. Antioxidant compounds: ....................................................................................... 18 2.12. Liver diseases:......................................................................................................... 18 2.12.1. Fascioliasis: ............................................................................................................... 18 2.12.2. Hepatitis: ................................................................................................................... 18 2.12.3. Alcoholic Liver disease: ............................................................................................ 19 2.12.4. Fatty Liver disease (Hepatic Steatosis): .................................................................... 19 2.12.5. Cirrhosis: ................................................................................................................... 19 2.12.6. Primary Liver Cancer: ............................................................................................... 19 2.12.7. Primary Biliary Cirrhosis: ......................................................................................... 19 2.12.8. Primary Sclerosing Cholangitis:................................................................................ 19 V 2.12.9. Centrilobular Necrosis of Liver: ............................................................................... 20 2.12.10. Budd– Chiari Syndrome: ....................................................................................... 20 2.12.11. Hereditary diseases: .............................................................................................. 20 2.12.12. Transthyretin-related hereditary amyloidosis:....................................................... 20 2.12.13. Gilbert's syndrome: ............................................................................................... 20 2.13. Spirulina keeps the Liver healthy: ....................................................................... 21 2.14. Liver Enzymes: ....................................................................................................... 21 2.15. Fibrosis: ................................................................................................................... 21 2.16. Evidence-Based Human Applications .................................................................. 22 2.16.1. Spirulina and Chronic Fatigue: ................................................................................. 22 2.16.2. Allergy, Rhinitis, and Immunomodulation: .............................................................. 22 2.16.3. Antiviral Applications: .............................................................................................. 23 2.16.4. Cholesterol-Lowering Effects and Effects on Diabetes: ........................................... 24 2.16.5. Anticancer Effects: .................................................................................................... 25 2.16.6. Chronic Arsenic Poisoning: ...................................................................................... 25 2.16.7. Antioxidant Effects: .................................................................................................. 26 2.17. General quality and safety assurance for Spirulina: ........................................... 26 3. Chapter three: Materials and Methods: ....................................................................... 29 3.1. Cultivation of Spirulina............................................................................................ 29 3.2 Preparation of Spirulina powder ............................................................................. 30 3.3. Chemical analysis of Spirulina Powder .................................................................. 30 3.4. Experimental animals: ............................................................................................. 30 3.5. Experimental design: ............................................................................................... 33 3.6. Drugs and Chemicals ............................................................................................... 34 3.6.1. D-galactosamine (D-GaIN): ............................................................................................ 34 3.6.2. Butylated hydroxytoluene (BHT): .................................................................................