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Name in Thesis ESTROGENIC ACTIVITY OF COSTUS SPECIOSUS (Koen.) Sm. EXTRACT IN FEMALE RATS Wanwisa Lijuan A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in Environmental Biology Suranaree University of Technology Academic Year 2010 ฤทธิ์การเปนเอสโตรเจนของสารสก ัดจากเอื้องหมายนา (Costus speciosus (Koen.) Sm.)ในหนูแรทเพศเมีย นางสาววันวิสาข ลิจวน วิทยานิพนธนี้เปนสวนหนงของการศึ่ ึกษาตามหลักสูตรปริญญาวิทยาศาสตรดุษฎีบัณฑิต สาขาวิชาชีววทยาสิ ิ่งแวดลอม มหาวิทยาลัยเทคโนโลยีสุรนารี ปการศึกษา 2553 ESTROGENIC ACTIVITY OF COSTUS SPECIOSUS (Koen.) Sm. EXTRACT IN FEMALE RATS Suranaree University of Technology has approved this thesis submitted in partial fulfillment of the requirements for the Degree of Doctor of Philosophy. Thesis Examining Committee ________________________________ (Assoc. Prof. Dr. Yupaporn Chaiseha) Chairperson ________________________________ (Assoc. Prof. Dr. Sajeera Kupittayanant) Member (Thesis Advisor) ________________________________ (Asst. Prof. Dr. Nuannoi Chudapongse) Member ________________________________ (Asst. Prof. Dr. Chusri Talubmook) Member ________________________________ (Asst. Prof. Dr. Griangsak Eumkeb) Member ________________________________ ________________________________ (Prof. Dr. Sukit Limpijumnong) (Assoc. Prof. Dr. Prapun Manyum) Vice Rector for Academic Affairs Dean of Institute of Science วันวิสาข ลิจวน : ฤทธิ์การเปนเอสโตรเจนของสารสกัดจากเอื้องหมายนา (Costus speciosus (Koen.) Sm.) ในหนแรทเพศเมู ีย (ESTROGENIC ACTIVITY OF COSTUS SPECIOSUS (Koen.) Sm. EXTRACT IN FEMALE RATS) อาจารยที่ปรึกษา : รองศาสตราจารย สัตวแพทยหญิง ดร.ศจีรา คุปพิทยานันท, 240 หนา. เอื้องหมายนา (Costus speciosus (Koen.) Sm. เปนพืชสมุนไพรพบทั่วไปในแถบเอเชียมี สรรพคุณเปนยาแผนโบราณใชในการรักษาโรคหลายชนิด เอื้องหมายนาประกอบดวยไฟโต- เอสโตรเจนหลายชนิดรวมทั้งไดออสจีนินและบีตาซิโตสเตอรอล วัตถุประสงคหลักของการศึกษานี้ คือศึกษาฤทธิ์การเปนเอสโตรเจนของสารสกัดจากเหงาและตนเอื้องหมายนาในหนูแรทเพศเมียโดย ศึกษา 1) ผลตอมดลูก ชองคลอด และเตานม 2) ผลตอระดับเอสตราไดออลและลิพิด โปรไฟล 3) ผลตอการฝงตัวและ 4) ผลตอการหดตัวของมดลูกและเปรียบเทียบผลกับสารมาตรฐาน พรอมทั้ง ศึกษากลไกการออกฤทธิ์ทางสรีรวิทยา ผลการศึกษาพบวาในสารสกัดจากเอื้องหมายนามีไดออส จีนินและบีตาซิโตสเตอรอลเชนเดียวกับไฟโตเอสโตรเจนอื่นๆ การศึกษาฤทธิ์พบวาการปอนสาร สกัดจากเอื้องหมายนาทั้งเหงาและตนขนาดต่ําและสูง (500 และ 1000 มก./กก.นน.) สามารถเพิ่ม น้ําหนักมดลูก เพิ่มความหนาของเยื่อบุชองคลอด เพิ่มจํานวนกระเปาะสรางน้ํานมและทอของเตา นมในหนูตัดรังไข สารสกัดจากเอื้องหมายนาไมเพิ่มระดับเอสตราไดออลแตลดระดับคลอ เรสเตอรอลและคลอเรสเตอรอลชนิดไมดี ที่เกิดจากการมีฤทธิ์คลายเอสโตรเจนของสารสกัดซึ่งไม เกี่ยวของกับการออกฤทธิ์ผานโกนาโดโทรปนพิทูอิตารียโอวาเรียนแอกซ ิส นอกจากนี้พบวาสาร สกัดเอื้องหมายนาทั้งเหงาและตนมีฤทธิ์ตอตานการฝงตัวในหนูทองระยะกอนการฝงตัว โดยเอื้อง หมายนามีผลไปรบกวนสมดุลระหวางโปรเจสเตอโรนและเอสโตรเจนตลอดจนไปเพิ่มการหดตัว ของมดลูก เมื่อศึกษาผลของสารสกัดเอื้องหมายนาตอสรีรวิทยาการหดตัวของมดลูกพบวาสารสกัด จากเหงาและตนเอื้องหมายนาสามารถเพิ่มการหดตัวของมดลูกในหนูปกติและหนูตัดรังไขได โดย มีฤทธิ์สูงสุดที่ 10 และ 30 มก./100 มล. ตามลําดับ โดยกลไกการออกฤทธิ์ไมไดเกิดจากการมีฤทธิ์ เปนเอสโตรเจนแตเกิดจากการเพิ่มการหดตัวซึ่งเกิดจากการเขาสูเซลลของแคลเซียมผานแอลไทป แคลเซียมชาแนลและการหลั่งแคลเซียมจากซาโคพลาสมิคเรติคูลัม สรุปไดวาเอื้องหมายนาทั้งเหงา และตนมีฤทธิ์การเปนเอสโตรเจน สาขาวิชาชีววิทยา ลายมือชื่อนักศึกษา__________________________ ปการศึกษา 2553 ลายมือชื่ออาจารยที่ปรึกษา____________________ ลายมือชื่ออาจารยที่ปรึกษารวม_________________ WANWISA LIJUAN : ESTROGENIC ACTIVITY OF COSTUS SPECIOSUS (Koen.) Sm. EXTRACT IN FEMALE RATS. THESIS ADVISOR : ASSOC. PROF. SAJEERA KUPITTAYANANT, Ph.D. (DVM) 240 PP. COSTUS SPECIOSUS (Koen.) Sm./ ESTROGEN/ RAT/ UTERUS/ VAGINA/ MAMMARY GLAND/ OVARIECTOMIZED RAT/ CONTRACTION/ CALCIUM Costus speciosus (Koen.) Sm. is a medicinal plant widely distributed in Asia. The plant is traditionally used in the treatment of various ailments. C. speciosus contains phytoestrogens including diosgenin and β-sitosterol. The main aim of this study was to study estrogenic activities of C. speciosus rhizome and stem extracts in female rats by investigating 1) the effects on uterus, vagina and mammary gland, 2) the effects on serum estradiol level and lipid profile, 3) the effects on implantation, and 4) the effects on uterine contraction and compared the effects to known compounds. The underlying mechanisms of the extracts were also investigated. The results revealed that C. speciosus extracts contained diosgenin and β-sitosterol as well as other phytoestrogens. The data showed that administered orally of the extracts both low and high doses (500 and 1000 mg/kg B.W.) increased relative uterine weight, vaginal epithelium thickness and mammary gland alveoli and ducts. The extracts did not increase serum estradiol level but decreased total cholesterol and low-density lipoprotein cholesterol. These estrogenic effects were not involved with gonadotropin- pituitary-ovarian axis. In addition, the extracts had anti-implantation effects during pre-implantation periods. These occurred via an alteration of progesterone and III estrogen balance as well as an increase in uterotonic activity. The investigation of physiological effects of C. speciosus extracts on uterine contractility exhibited that C. speciosus rhizome and stem extracts are potent stimulators of the uterus in both non- pregnant and ovariectomized rats as they increased spontaneous contraction with a maximum effect of 10 and 30 mg/100 mL, respectively. The mechanisms of action were due to non-estrogen effect, but increasing contraction via Ca2+ entry on L-type calcium channel and sarcoplasmic reticulum Ca2+ release. In conclusion, the study clearly showed that both C. speciosus rhizome and stem have estrogenic activity. School of Biology Student’s Signature_____________________ Academic Year 2010 Advisor’s Signature_____________________ Co-advisor’s Signature__________________ ACKNOWLEDGEMENTS First of all, I would like to gratefully thank my thesis advisor, Assoc. Prof. Dr. Sajeera Kupittayanant, who gave valuable advice, suggestions and comments to my work. With gratefulness and respect, all the kindness and support I have been received from her will be in my memory. I would like to thank my co-advisor, Asst. Prof. Dr. Nuannoi Chudapongse, and the thesis committee, Assoc. Prof. Dr. Yupaporn Chaiseha, Asst. Prof. Dr. Chusri Talubmook and Asst. Prof. Dr. Griangsak Eumkeb, for their valuable suggestions. I would also like to thank all the lecturers of Suranaree University of Technology (SUT), who had taught me in all courses during my studies. I thank all staff of the Center for Scientific and Technological Equipment as well as staff at the Animal House, the Physiological Laboratory and the Biological Laboratory for their technical support. Thanks all of my friends for good things we have shared and thanks to sisters and brothers in the Reproductive Laboratory for their friendly help during my studies. I would like to thank SUT for financial support (Intelligent Graduate Scholarship). Special thanks to my parents for their kind support, understandings, encouragement and love. Wanwisa Lijuan CONTENTS Page ABSTRACT IN THAI................................................................................................... I ABSTRACT IN ENGLISH ..........................................................................................II ACKNOWLEDGEMENTS........................................................................................ IV CONTENTS..................................................................................................................V LIST OF TABLES.....................................................................................................XV LIST OF FIGURES ...............................................................................................XVIII CHAPTER I INTRODUCTION............................................................................................. 1 1.1 Female Reproductive Cycle......................................................................... 1 1.1.1 Ovarian Cycle.................................................................................... 2 1.1.2 Uterine Cycle..................................................................................... 3 1.2 Implantation and Pregnancy......................................................................... 5 1.3 Menopause................................................................................................... 6 1.3.1 The Causes of Menopause................................................................. 8 1.3.2 Symptoms of the Menopause ............................................................ 9 1.3.3 Hormone Replacement Therapy...................................................... 10 1.4 Estrogen and Its Functions......................................................................... 12 1.4.1 Effects of Estrogen on Female Reproductive System..................... 12 1.4.2 Effects of Estrogen on Lipid Profile ............................................... 13 VI CONTENTS (Continued) Page 1.4.3 Effects of Estrogen on Fertility Control.......................................... 14 1.5 Herbal Medication...................................................................................... 15 1.6 Costus speciosus (Koen.) Sm..................................................................... 18 1.7 Aims .........................................................................................................19 1.8 References.................................................................................................
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