The Effects of Epimedii Herba on a Hypothyroidism Rat Model induced by PTU(6-Propyl, 2-thiouracil)

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(1)ISSN 2093-6966. Journal of Pharmacopuncture. 13 http://dx.doi.org/10.3831/KPI.2011.14.4.013. Original Article. 淫羊藿이 실험적으로 유발된 갑상선 기능저하증 동물모델에 미치는 영향과 안전성. Received : Sep 28, 2011 Revised : Nov 29, 2011. The Effects of Epimedii Herba on a Hypothyroidism Rat Model induced by PTU(6-Propyl, 2-thiouracil).. Accepted : Nov 30, 2011 Min-Jin Hong, Byung-Choel Lee, Young-Min Ahn, Se-Young Ahn. KEY WORDS: Epimedii Herba; 6-Propyl, 2-thiouracil; Hypothyroidism. Department of Internal medicine, Kyung Hee University College of Oriental Medicine, Seoul, Korea. Abstract Objectives: The symptoms of hypothyroidism are fatigue, cold intolerance, arthralgia, muscle cramps, dry skin and etc. Although hypothyroidism is a relatively common endocrinical disease, we do have many difficulties treating it effectively. The symptoms of hypothyroidism are similar to those of Yang-deficiency in Oriental medicine. Epimedii Herba is a popular herb that has the effect of tonifying the kidney and strengthening Yang in Oriental medicine. In this study, we investigated the therapeutic effects of Epimedii Herba on a hypothyroidism rat model induced by PTU(6-Propyl, 2thiouracil). Methods: 24 two-month-old Spargue-Dawley(SD) rats were divided into 4 groups: 1) normal(n=6), 2) PTU-induced hypothyroidism control(n=6), 3) hypothyroidism rat treated with Epimedii Herba(n=6), 4) hypothyroidism rat treated with levothyroxine(n=6). PTU was administered for 4 weeks, Epimedii Herba and levothyroxine was administered for 2 weeks after PTU was initiated for a total duration of 2 week. At the end of the experiment, blood samples from all the rats were taken from their hearts and were analyzed. Results: In comparison with normal group, the PTU-induced control group significantly showed hypothyroidism with low T3, T4 and high TSH. In Epimedii Herba group, T4 was significantly increased(p<0.05). There was no significant difference in TSH between the Epimedii Herba treatment group and the control group. And no significant differences were observed in biochemical labs and weight between the Epimedii Herba group and the control group. Conclusions: These results suggest that Epimedii Herba could help thyroid cells to produce thyroid hormones. And no significant side effects related with Epimedii Herba were found, suggesting that it is safe to administer. According to these results, Epimedii Herba may be a safe alternative medicine for hypothyroidism.. cc This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.. 2011 Korean Pharmacopuncture institute. Corresponding Author Se-Young Ahn, Department of Internal medicine VI, Kyung Hee University, Hoegi-dong, Dongdaemun-gu, Seoul 446-701, Korea Tel: +82-2-958-9153, Fax: +82-2-958-9158 E-mail: [email protected] http://pharmacopuncture.co.kr.

(2) 14. Journal of Pharmacopuncture 제 14권 제 4호( 2011년 12월). I. 서론. cold intolerance. PTU Rat. II. 재료 및 방법 1. Epimedii Herba 1,500 80 heating mantle 2 500 applicator filter Rotary evaporator Model NE-1 Japan 1 1 10 95 2 1,000. 14,000rpm. 20 0.2. Levothyroxine TSH. 70 19.7. 2. PTU Epimedii Herba Berberidaceae Epimedium bervicornum Maxim, E.sagittatum S. et Z. Maxim E. koreanum Nakai. 2 rat. 200 20 Korea 40~70. Sprague-Dawley. male. cage. 1 6-Propyl, 2-thiouracil water supplement Ca P3755, 10g, sigma 400 H2O 40 rat 100 0.2 day 4.

(3) 15. 3.. 4.. SD male rat 6 Control group Thyroxine LT4 Normal group Propyl, 2-thiouracil. Normal group 1. sampling. PTU 64 PTU. 2. 500 Zonde 1 Control group. CAS 2.5D, Korea rat bowl rat. rat. 1 2 PTU. 2 LT4 PTU 2 Levothyroxine CA T2376, 1g, sigma 0.5N NaOH isotonic saline 250 500 rat 100 0.1 day 2 Table 1, Fig. 1 Table 1. Structure of the Experimenta Group No. of Rat. PTU. Levothyroxine. Treatment. Normal. 6. none. none. No-treatment. Control. 6. 0.2 /100g(s.c.). none. Saline(p.o.). LT4b. 6. 0.2. /100g(s.c.) 0.1. /100g(i.p.). Saline(p.o.). Epimediic. 6. 0.2. /100g(s.c.). none. Epimedii Herba 500 / (p.o.). 5. 4 rat 10cc 3,000rpm 20 -40 AST Aspartate Transaminase ALT Alanine Transaminase ALP Alkaline Phosphatase BUN Blood Urea Nitrogen Creatinine, Glucose, Albumin, Protein, Total Cholesterol, HDL-Cholesterol, LDL-Cholesterol, Triglyceride. 6.. T3, T4. TSH. a. Each parenthesis represents the route of administration. s.c.,subcu-. taneous injection; i.p.,intraperitoneal; p.o.,oral administration. LT4 b, Levothyroxine 0.1 500 /. /100g treatment group. Epimediic, Epimedii Herba. treatment group.. Figures & Tables start. 1week. 2week. T3, T4 TSH Rodent T3 ELISA Enzyme-Linked Immunosorbent Assay test kit Endocrine technologies Inc., USA. Rodent T4 ELISA test kit Rodent TSH ELISA test kit 3,000rpm 20 -40. 3week. 4week. T3. PTU(6-propyl, 2-thiouracil) (s.c.). Epimedii Herba (p.o.) Levothyroxine (i.p.). sampling. Fig. 1. The experimental schedule for this study. T3-antibody microtiter wells 50 standard T3 solution 100 T3 HRP Horseradish Peroxidase conjugate, 100 TBM color solution, 50 2N HCl stop solution ELISA reader 450nm T3 T4 TSH T4-antibody TSH-antibody microtiter wells 100 standard T4 TSH solution.

(4) Journal of Pharmacopuncture 제 14권 제 4호( 2011년 12월). 16. 100 T4 TSH enzyme conjugate, 100 TBM color solution, 50 2N HCl stop solution ELISA reader 450nm T4 TSH. 5. LT4 7.. Table 2 Table 2. Body Weight of Each Experimental Group. rat 70. 80. 10 95. 100. 0week(g). 100. 95 80 rehydration matoxyline-Eosin HE. 2week(g). 3week(g). 4week(g). Normal. 238.3 3.98. 252.8 8.13. 306.0 10.88. 339.0 26.42. 347.5 41.05. Control. 240.3 6.35. 280.5 5.39. 295.2 2.14. 262.2 11.27. 291.8 37.04. LT4. 241.3 4.55. 277.2 3.97. 301.5 6.38. 266.5 9.01. 303.5 9.33. Epimedii. 244.0 2.90. 284.3 6.83. 302.2 19.30. 313.7 20.53. 312.5 24.55. microtome 4 gelatin coated slide. 1week(g). xylene 70 He2. Olympus BX-50,. Olympus Optical, Tokyo, Japan 400. rat ALP, AST, ALT. 8.. LT4 69.2 12.25IU L. GraphPad PRISM statistical package ver 4.03, Graphpad software inc., San Die go, USA one-way analysis of variance ANOVA Tuckey’s post-hoc test mean S.D. Twotailed p value p 0.05. AST. Table 3 Table 3. Biochemical Analysis of Alkaline Phosphatase, Aspartate Transaminase and Alanine Transaminase in Each Experimental Group ALP(IU/L). AST(IU/L). ALT(IU/L). Normal. 161.7. 32.68. 123.3. 31.68. 52.5. 11.40. Control. 128.2. 26.87. 117.5. 42.44. 46.0. 3.69. LT4. 120.0. 29.87. 69.2. 12.25*. 36.2. 10.68. Epimedii. 101.2. 22.19. 5.10. 35.6. 7.64. III. 결과 1.. LT4 117.5 42.44IU/L p 0.05 ALP, ALT. 73.9. *Significantly different from the control group at p 0.05. rat 1. 4. 3..

(5) 17. BUN. LT4 LT4. Creatinine. BUN Creatinine 0.5 0.04 p 0.05. 0.4 0.08. Total Cholesterol HDLCholesterol, LDL-Cholesterol, Triglyceride Triglyceride 43.3 12.66 85.5 35.21 p 0.05 HDL-Cholesterol, LDL-Cholesterol, Total Cholesterol. Table 4. LT4. Table 4. Biochemical Analysis of Blood Urea Nitrogen and Creatinine in Each Experimental Group BUN( /. ). Cr(. / ). Normal. 17.2 1.77. 0.5. 0.08. Control. 17.5 2.38. 0.5. 0.04. LT4. 17.0 2.88. 0.4 0.08*. 20.00. 0.5. Epimedii. 1.42. Total Cholesterol LT4 9.65 76.8 13.70 p 0.05 Table 6. 53.5. Table 6. Biochemical Analysis of HDL-Cholesterol, LDLCholesterol, Total Cholesterol and Triglyceride in each experimental group HDL-Chol ( / ). LDL-Chol ( / ). Normal. 28.8. 5.50. 14.5. 6.83. 66.8. 17.94. 85.5 35.21. Control. 31.7. 5.62. 15.0. 3.85. 76.8. 13.70. 43.3 12.66#. LT4. 26.4. 3.97. 9.7. 2.16. 53.5. 9.65*. 53.2 15.69. Epimedii. 23.7. 3.21. 18.7. 3.49. 62.9. 11.05. 30.6. 0.05. *Significantly different from the control group at p 0.05. 4. #. T-Chol ( / ). Tg( / ). 9.02. Significantly different from the normal group at p 0.05. *Significantly different from the control group at p 0.05. 103.0 32.75 122.2 13.38 6. LT4 rat Table 5 Table 5. Biochemical Analysis of Glucose in each experimental group. Glucose( / ) Normal. 122.2. 13.38. Control. 103.0. 32.75. LT4. 125.3. 30.12. Epimedii. 132.1. 17.02. LT4 Albumin, Total Protein Table 7 Table 7. Biochemical analysis of Albumin and Total Protein in each experimental group Albumin(g/ ). 5.. Total Protein(g/. Normal. 1.9. 0.29. 5.0. 1.15. Control. 1.8. 0.20. 5.2. 0.96. LT4. 1.4. 0.29. 3.9. 0.57. Epimedii. 1.6. 0.15. 4.7. 0.60. ).

(6) Journal of Pharmacopuncture 제 14권 제 4호( 2011년 12월). 18. 7. PTU rat T3, T4. TSH. Thyroglobulin T3. 0.05. 0.8 0.13 p 0.001 4.3 0.93 p 0.001 1.7 0.57 IU p 0.001. IU. 0.5. colloid epithelial cell. T4. TSH. 1.0 0.09 LT4. TSH. 5.1 1.03 epithelial cell. T3. LT4. T4. LT4. 6.1 1.53. 1.0 0.09 p 0.001. Fig. 2. T4. 1.3 0.30 p 0.05 1.4 0.35 IU. TSH. LT4 5.1 1.03 IU p 0.001. Fig. 2. Histology of thyroid gland of Normal, Control, LT4, Epimedii-treated Hypothyroidism rats inducued by PTU. Control, PTU; T4, PTU + T4; Epimedii, PTU + Epimedii.. Table 8 Table 8. Level of T3, T4 and TSH in each experimental group T3 (ng/ Normal Control. ###. 0.8. ). 0.13. 0.5 0.05. ###. T4 ( g/ 4.3. ). TSH( IU/. 0.93. 1.0 0.09. 1.7 0.57. ###. LT4. 0.5. 0.04. 6.1 1.53***. Epimedii. 0.6. 0.07. 1.3 0.30*. ). 5.1 1.03### 1.4. 0.35***. 4.0 0.81. Significantly different from the normal group at p 0.001. ***Significantly different from the control group at p 0.001 *Significantly different from the control group at p 0.05. 8. PTU. Rat. Hematoxyline-Eosin HE follicle. IV. 고찰.

(7) 19. PTU Rat T3, T4 T3. TSH T4 TSH TSH T3, T4 T3, T4 T3, T4 Epimedii Herba ridaceae bervicornum Maxim Z. Maxim. BerbeEpimedium E.sagittatum S. et E. koreanum Nakai. TSH. T4 TSH. T4. T4. T3 T3. TSH. T4. T3. T3 PTU 20. flavonoid icariin noricariin lignan cresyl alcohol, flavonol glycoside korepimedoside A B. T4. T3 T4. PTU TSH LT4 negative feed back. TSH TSH.

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