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Bouea Macrophylla

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Bouea Macrophylla BIOLOGICAL ACTIVITIES OF THAI TRADITIONAL REMEDY CALLED PRASACHANDAENG AND ITS PLANT INGREDIENTS BY MISS ALISA SANGPHUM A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN APPLIED THAI TRADITIONAL MEDICINE FACULTY OF MEDICINE THAMMASAT UNIVERSITY ACADEMIC YEAR 2016 COPYRIGHT OF THAMMASAT UNIVERSITY Ref. code: 25595611031526QZI BIOLOGICAL ACTIVITIES OF THAI TRADITIONAL REMEDY CALLED PRASACHANDAENG AND ITS PLANT INGREDIENTS BY MISS ALISA SANGPHUM A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN APPLIED THAI TRADITIONAL MEDICINE FACULTY OF MEDICINE THAMMASAT UNIVERSITY ACADEMIC YEAR 2016 COPYRIGHT OF THAMMASAT UNIVERSITY Ref. code: 25595611031526QZI (1) Thesis Title BIOLOGICAL ACTIVITIVES OF THAI TRADITIONAL REMEDY CALLED PRASACHANDAENG AND ITS PLANT INGREDIENTS Author Miss Alisa Sangphum Degree Master of Science Major/Faculty/University Applied Thai Traditional Medicine Faculty of Medicine Thammasat University Thesis Advisor Associate Professor Arunporn Itharat, Ph.D. Academic Years 2016 ABSTRACT Prasachangdaeng remedy (PC) has long been used for relief of fever and internal heat. It is composed with 12 plants as Chan Daeng (Dracaena loureiri Gagnep.) stems, Mueng Kon (Helicia terminalis Kurz.) roots, Ma Now (Citrus aurantifolia Swing.) roots, Ma Prang Wan (Bouea macrophylla Griff.) roots, Proh Horm (Kaempferia galanga L.) rhizome, Kot Hua Bua (Ligusticum chuanxiong Hort.) rhizome, Chan Tet (Myristica fragrans Houtt.) stems, Fang Saeng (Caesalpinia sappan L.) stems, Bua Luang (Nelumbo nucifera Gaertn.) pollens, Boon Ngak (Mesua ferrea L.) flowers, Sa Ra Pee (Mammea siamensis Kosterm.) flowers, and Ma Li (Jasminum sambac L.) flowers. It is in the National List of Essential Medicines in Thailand, but there is no report for anti-inflammatory activity of this remedy. Some plant ingredients in this remedy were investigated for antibacterial and anti-inflammatory activities but there is no report of PC and some plant ingredients in PC remedy. Thus, the objectives of this research were to study on antibacterial and anti-inflammatory activities of PC remedy extracts by different extraction method and its plant ingredients. Raw material of its plant ingredients from PC remedy were tested for standardization following Thai Herbal Pharmacopoeia (THP). All plant ingredients of PC remedy were accepted by standard values. PC remedy was also accepted by THP Ref. code: 25595611031526QZI (2) criteria (the moisture content of PC remedy was 6.74±0.67%, total ash as 5.69±0.08% and acid insoluble ash as 1.78±0.03%). PC remedy was extracted by different methods by maceration in 95% ethanol and decoction. The yields of PC ethanolic extracts (PCE) were higher than water extract (PCW) (17.28% and 3.35%, respectively). Each plant ingredients were also extracted by 95% ethanol. The ethanolic extract of Dracaena loureiri Gagnep. stems exhibited the highest yield and Myristica fragrans Houtt. stems exhibited the lowest yield (22.76% and 1.24%, respectively). For decoction; Ligusticum chuanxiong Hort. Rhizomes water extract exhibited the highest yield and Myristica fragrans Houtt. stems exhibited the lowest yield (27.29% and 0.85%, respectively) Antibacterial activity were evaluated by disc diffusion method to determine the inhibition zone and by using microtitre plate-based antibacterial assay to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values and tested against three of gram positive bacteria; Staphylococcus aureus (ATCC 25923), Staphylococcus aureus MRSA (DMST 2065) and Streptococcus pyogenes (ATCC 19615) and four types of gram negative bacteria; Escherichia coli (ATCC 25922), Klebsiella pneumonia (ATCC 700603), Salmonella typhimurium (DMST 22842) and Shigella dysenteriae (DMST 151161). The PCE and PCW showed antibacterial activity against S.aureus MRSA (MIC = 0.312 mg/ml, MBC = 2.5 mg/ml and MIC = 1.25 mg/ml, MBC = 1.25 mg/ml, respectively). The inhibition zones of all plant extracts were in the range of 7 to 26 mm. The ethanolic and water extracts of Caesalpinia sappan L. stems showed the largest inhibition zone against S.aureus MRSA as 26 and 20 mm., respectively. The ethanolic extract of Mammea siamensis Kosterm. flowers exhibited the best gram positive bacteria activity against S.aureus MRSA with MIC and MBC values of 0.004 and 0.019 mg/ml., respectively. The ethanolic extract of Caesalpinia sappan L. stems exhibited the best inhibitory gram negative bacteria activity against S.dysenteriae with MIC and MBC values of 0.156 and 0.156 mg/ml. Most of water extracts of PC remedy and its plant ingredients had no antibacterial activity against all strains. Neither ethanolic nor water extracts of PC remedy and its plant ingredients were able to inhibit gram negative K. pneumonia. except, the ethanolic and water extracts of Caesalpinia sappan L. stems exhibited the inhibitory gram negative bacteria activity against K. pneumonia. with Ref. code: 25595611031526QZI (3) MIC value = 1.25 mg/ml, MBC value = 5 mg/ml, and MIC value = 2.5 mg/ml, MBC value = 5 mg/ml., respectively. Anti-inflammatory activity was evaluated by inhibition Nitric Oxide (NO) production which induced by Lipopolysaccharide (LPS) from murine macrophages cell lines (RAW264.7). The results found that the PCW showed better anti-inflammatory activity than PCE (IC50 value of 16.87±2.51 and 39.70±1.48 µg/ml.,respectively). The ethanolic and water extracts of Caesalpinia sappan L. stem showed the highest anti- inflammatory activity with IC50 value 5.42±0.24 µg/ml and 7.60±0.28 µg/ml, respectively. The stability test by accelerated condition (40ºC, 75%RH for 6 months) of Prasachandaeng extract found that the water extract of PC remedy was stable in anti- inflammatory activity by inhibitory NO production from RAW264.7 cell lines for at least 8 months. In conclusion, the PCE showed good antibacterial activities while the PCW showed good anti-inflammatory activity. The ethanolic extract of Mammea siamensis Kosterm. flowers showed the effective antibacterial activities against MRSA and the ethanolic (FSE) and water (FSW) extract of Caesalpinia sappan L. stems showed the effective anti-inflammatory activity. Furthermore, the FSW and FSE also showed good antibacterial activities. Thus, these results support to use the PCE for treat infection fever and MRSA infection and using PCW for treat fever from inflammation. Thus, these results are related on using PC in Thai Traditional Medicine scripture. Keywords: Prasachandaeng, Thai Traditional remedy, Antibacterial, Anti- inflammatory Ref. code: 25595611031526QZI (4) หัวข้อวิทยานิพนธ์ ฤทธิ์ทางชีวภาพของต ารับยาไทยชื่อยาประสะจันทน์แดงและ สมุนไพรในต ารับ ชื่อผู้เขียน นางสาวอลิษา แสงพุ่ม ชื่อปริญญา วิทยาศาสตรมหาบัณฑิต สาขา/คณะ/มหาวิทยาลัย สาขาวิชาการแพทย์แผนไทยประยุกต์ คณะแพทยศาสตร์ มหาวิทยาลัยธรรมศาสตร์ อาจารย์ที่ปรึกษาวิทยานิพนธ์ รองศาสตราจารย์ ดร.อรุณพร อิฐรัตน์ ปีการศึกษา 2559 บทคัดย่อ ต ารับยาประสะจันทน์แดงเป็นต ารับยาสมุนไพรไทย ซึ่งมีการใช้รักษาอาการไข้ตัว ร้อน และบรรเทาอาการร้อนใน กระหายน ามายาวนาน ประกอบด้วยสมุนไพรทั งหมด 12 ชนิด ได้แก่ แก่นจันทน์แดง (Dracaena loureiri Gagnep.), รากเหมือดคน (Helicia terminalis Kurz.), ราก มะนาว (Citrus aurantifolia Swing.), รากมะปรางหวาน (Bouea macrophylla Griff.), หัว เปราะหอม (Kaempferia galanga L.), โกฐหัวบัว (Ligusticum chuanxiong Hort.), แก่น จันทน์เทศ (Myristica fragrans Houtt.), แก่นฝางเสน (Caesalpinia sappan L.), เกสรบัวหลวง (Nelumbo nucifera Gaertn.), ดอกบุนนาค (Mesua ferrea L.), ดอกสารภี (Mammea siamensis Kosterm.) และดอกมะลิ (Jasminum sambac L.) ต ารับยาประสะจันทน์แดงเป็นต ารับ ที่อยู่ในบัญชียาหลักแห่งชาติ แต่อย่างไรก็ตามต ารับยาประสะจันทน์แดงนั น ยังไม่มีรายงานผลการวิจัย ถึงฤทธิ์ต้านการอักเสบ ในส่วนของสมุนไพรในต ารับบางชนิด มีงานวิจัยถึงฤทธิ์ต้านเชื อแบคทีเรียและ ฤทธิ์ต้านการอักเสบ แต่สมุนไพรบางตัวในต ารับไม่มีรายงานวิจัยถึงฤทธิ์ดังกล่าว ดังนั นวัตถุประสงค์ ของการวิจัยนี เพื่อศึกษาถึงฤทธิ์ต้านเชื อแบคทีเรียและฤทธิ์ต้านการอักเสบของสารสกัดด้วยเอทานอล และสารสกัดด้วยน าของต ารับยาประสะจันทน์แดงและสมุนไพรไทยในต ารับ ต ารับยาประสะจันทน์แดงและสมุนไพรในต ารับแต่ละชนิดผ่านเกณฑ์มาตรฐานการ ควบคุมคุณภาพมาตรฐานยาสมุนไพรไทย โดยต ารับยาประสะจันทน์แดงผ่านเกณฑ์มาตรฐาน ดังต่อไปนี ค่าบ่งชี ปริมาณน า 6.74±0.67% ปริมาณเถ้าทั งหมด 5.69±0.08% และปริมาณเถ้าที่ไม่ ละลายในกรด 1.78±0.03% ต ารับยาประสะจันทน์แดงและสมุนไพรแต่ละชนิด น ามาสกัดด้วยวิธีการหมักด้วยเอ ทานอลความเข้มข้น 95% และสกัดด้วยวิธีการต้มน า สารสกัดด้วยเอทานอลได้ผลผลิตร้อยละมากกว่า Ref. code: 25595611031526QZI (5) สารสกัดด้วยน า เท่ากับ 17.28% และ 3.35% ตามล าดับ สมุนไพรแต่ละชนิดที่สกัดด้วยเอทานอล ความเข้นข้น 95% พบว่าแก่นจันทน์แดงได้ผลผลิตร้อยละมากที่สุด และแก่นจันทน์เทศได้ผลผลิตร้อย ละน้อยที่สุดเท่ากับ 22.76% และ 1.24% ตามล าดับ ในส่วนของสมุนไพรที่สกัดด้วยการต้มน า พบว่า โกศหัวบัวได้ผลผลิตร้อยละมากที่สุด และแก่นจันทน์เทศได้ผลผลิตร้อยละน้อยที่สุด เท่ากับ 27.29% และ 0.85% ตามล าดับ การศึกษาฤทธิ์ต้านแบคทีเรียด้วยวิธี Disc diffusion เพื่อหาเส้นผ่าศูนย์กลางของ Inhibition Zone และวิธี Microtitre plate-based antibacterial assay เพื่อหาค่าความเข้มข้น ต่ าสุดที่สามารถยับยั งการเจริญเติบโตของเชื อแบคทีเรีย (MIC) และค่าความเข้มข้นต่ าสุดที่สามารถ ฆ่าเชื อแบคทีเรีย (MBC) โดยทดสอบกับเชื อแบคทีเรียแกรมบวก 3 สายพันธุ์ คือ Staphylococcus aureus (ATCC 25923), Staphylococcus aureus MRSA (DMST 2065) และ Streptococcus pyogenes (ATCC 19615) และเชื อแบคทีเรียแกรมลบ 4 สายพันธุ์ คือ Escherichia coli (ATCC 25922), Klebsiella pneumonia (ATCC 700603), Salmonella typhimurium (DMST 22842) และ Shigella dysenteriae (DMST 151161) พบว่าสารสกัดด้วยเอทานอลและสารสกัดด้วยน
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