Herbal Formula Science (대한한의학방제학회지)

The Korean Medicine Society for the Herbal Formula Study (대한한의학방제학회)
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Ephedra has anti-fibrogenic effects by inhibiting the TGF-β/Smad pathway in LX-2 cells

마황(麻黃) 열수 추출물의 TGF-β/Smad 경로 억제를 통한 간섬유화 억제효능

  • Jea Hyun Yoo (College of Korean Medicine, Daegu Haany University) ;
  • Sang Mi Park (College of Korean Medicine, Daegu Haany University) ;
  • Dae Hwa Jung (Department of Pharmaceutical Engineering, Daegu Haany University) ;
  • Sang Chan Kim (College of Korean Medicine, Daegu Haany University)
  • 유재현 (대구한의대학교 한의과대학) ;
  • 박상미 (대구한의대학교 한의과대학) ;
  • 정대화 (대구한의대학교 제약공학과) ;
  • 김상찬 (대구한의대학교 한의과대학)
  • Received : 2024.04.05
  • Accepted : 2024.05.23
  • Published : 2024.05.31
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Abstract

Objective : Ephedrae Herba (Ephedra) has been frequently used in the East Asian traditional medicine including Korea, China and Japan in the clinical treatment of asthma, cold and influenza etc. This study was performed to explore an anti-fibrogenic potential of Ephedra Herba water extract (EHE) using immortalized human hepatic stellate cell line, LX-2 cells. Methods : We examined the anti-fibrogenic effects of EHE on canonical pathway of transforming growth factor-β1 (TGF-β1) signaling in LX-2 cells. Cell viability was measured using the MTT assay. mRNA levels were detected by real-time PCR. Proteins expression were detected by Western blot. Results : Treatment of EHE 30 ㎍/ml did not show any cytotoxicity on LX-2 cells. Pre-treatment of EHE (30 ㎍/mL) significantly inhibited α-smooth muscle actin expression induced by TGF-β1. Additionally, EHE significantly decreased Smad2 and Smad3 phosphorylations, Smad binding element-driven luciferase activity and plasminogen activator inhibitor type 1 expression by TGF-β1. Furthermore, increases of matrix metalloproteinases 2 genes by TGF-β1 was also attenuated by EHE treatment. Conclusion : These results suggest that EHE has an ability to suppress fibrogenic process in activated HSC via inhibition of TGF-β1-TGFBR mediated canonical (Smad dependent) pathway.

Keywords

Acknowledgement

This study was supported by the National Research Foundation of Korea funded by Korea government (MSIP) (Grant No.2018R1A5A2025272)

References

  1. Lee SI. Herbology. Seoul:Suseowon. 1981:189-90.
  2. Hwang CH, Yun YG. Study on Application of Herba Ephedrae Main Blended Prescription from Dongeuybogam. The Koran Journal of Oriental Medical Prescription. 2005;13(2):16-92.
  3. Park BR, Lee SH, Han KM, Hwang JW, Kim HI, Baek SY. Development and Utilization of KASP Markers for the Identification of Three Types of Ephedra Herbs. Kor. J. Pharmacogn. 2022;53(4):226-33.
  4. Lee GS. The Anatomical Identification-key of Ephedrae Herba and its adulterant. Kor. J. Herbol. 2019;34(6):25-32.
  5. Ha TH, Kwon TW, Kim YK. Effects of Ephedra Herba ethyl-acetate fraction on high fat diet induced hyperlipidemic mice. Journal of Society of Preventive Korean Medicine 2014;18(2):101-13.
  6. Kim HJ, Kim EJ, Han YH. The Effect of Ephedra sinica Pharmacopuncture on Lipid Metabolism in an Experimental Mouse Model of Obesity. J. Int. Korean Med. 2016;37(4):579-90.
  7. Jeong JJ, Kim BW. The Effect of Ephedrae Herba Pharmacopuncture on Adipocyte Metabolism. Korean J. Orient. int. Med. 2008;29(1):80-9.
  8. Won CW, Jung YS, Yoon KH, Lee HY, Yoon MC, Kim BK, Park SD, Shin SS. Herba Ephedrae and Rhizoma Amorphophalli modulates visceral obesity in micro-CT of high fat induced obese male mice. The Koran Journal of Oriental Medical Prescription. 2008;16(2):205-17.
  9. Oh MJ, Lee CH, Kim HJ, Kim HR, Kim MS, Lee DY, Oh CH, Kim MS, Kim JS. The comparative studies on anti-obesity effects of Ephedrae Herba and Cyperi rhizoma in high fat diet fed mice. Herb. Formula Sci. 2016;24(2):108-23.
  10. Jo EH, Jo IJ, Park SJ, Jo SH, Park MC. Effects of Ephedra sinica (ES) Extract on the Ovalbumin-Induced Allergid Asthma in Mice. J Korean Med Ophthalmol Otolaryngol Dermatol. 2014;27(3):84-95.
  11. Ryu HC, Shin JG. A study on proper dosage of Ma-huang. J of KMediACS. 2013;5(1):101-11.
  12. Lee TH, Hsing LC, Yang CS, Kim LH, Seo ES, Jang IS. Adverse Effects of Ephedra According to Sasang Typology in Healthy Adults : ADouble-Blind Randomized Controlled Trial. Korean J. Orient. Int. Med. 2009:30(1)144-52.
  13. Lee TH. Literature investigation on the meaning of Mahwang, Gyeji, Sesin, Buja and Related Prescriptions. Herb. Formula Sci. 2018;26(3):261-5.
  14. Ming Y, Cheng S, Long W, Wang HL, Xu C, Liu X, Zhang Q, Zhao S, Zou X, Fan J, Wang L, Shen H. The Herbal Formula Granule Prescription Mahuang Decoction Ameliorated Chronic Kidney Disease Which Was Associated with Restoration of Dysbiosis of Intestinal Microbiota in Rats. Evid Based Complement Alternat Med. 2021:4602612.
  15. Dai H, Liu F, Qiu X, Liu W, Dong Z, Jia Y, Feng Z, Liu Z, Zhao Q, Gao Y, Zhang Z, Gao C, Sun S, Tian X, Liu B. Alleviation by Mahuang Fuzi and Shenzhuo Decoction in High Glucose-Induced Podocyte Injury by Inhibiting the Activation of Wnt/beta-Catenin Signaling Pathway, Resulting in Activation of Podocyte Autophagy. Evid Based Complement Alternat Med. 2020:7809427.
  16. Kang HJ, Ku TH, Kim GC. Hemodynamic Effects of Herbal Prescription Containing Ephedra on Weight Loss: A 3-Dimensional Radial Pulse Tonometry Device Study. J Korean Med Obes Res 2023;23(1):28-41.
  17. Zhai HQ, Zhang SF, Gao MC, Liu Y, Ou M, Meng FY, Wang YY. Effects of Herba Ephedra Sinicae and Fructus Schisandrae Chinensis on pathology of rats with bleomycin A(5)-induced idiopathic pulmonary fibrosis. Zhong Xi Yi Jie He Xue Bao. 2011;9(5):553-7.
  18. Zhuo Z, Nie J, Xie B, Wang F, Shi M, Jiang Y, Zhu W. A comprehensive study of Ephedra sinica Stapf-Schisandra chinensis (Turcz.) Baill herb pair on airway protection in asthma. J Ethnopharmacol. 2024;322:117614.
  19. Yoon KH, Lee HY, Jung YS, Seo BI, Park GY, Yoon MC, Shin SS. Modulation of obesity by Gyeongshingangjeehwan18 in ob/ob mice. Kor. J. Herbology 2010;25(3):1-9.
  20. Lee HY, Yoon KH, Seo BI, Park GY, Yoon MC, Shen ZB, Cui HH, Shin SS, Molecular biologic mechanism of obesity by GGEx18. Kor. J. Herbology 2011;26(1):65-74.
  21. Kim HJ, Kim JB. Effects of Mahwangimun-tang in Cholesterol Diet and Triton WR-1339-induced Hyperlipidemic Rat. Korean J. Oriental Physiology & Pathology. 2010;24(1):102-10.
  22. Kim KH, Kang GH, Lee YU, Kang S, Ji SH, Song JH, Kim CH, Lee SK. Treating COVID-19 with Qingfei Paidu Decoction with and without Ephedra Herba: A Retrospective Case Series. J. Int. Korean Med. 2022;43(2):264-73.
  23. Yun CS, Hong SH, Park MC, Hwang CY. The Effects of Bojungikgitang-gamibang Administration along with Mahwangshingungsan on the Rat Model with Allergic Rhinitis. The Journal of Korean Oriental Medical Ophthalmology & Otolaryngology & Dermatology 2008;21(3):111-23.
  24. Park JE, Hwang HS, Kim TY. Effects of Mahwanghangingamchosukgo-tang Extracts. Korean J. Oriental Physiology & Pathology. 2020;34(6):319-25.
  25. Jang HK, Heo DS. Anti-OsteoArthritic effects of Mahwangbuja-tang(Mahuangfuzi-tang) on Papain- Induced OsteoArthritic mice model. J Oriental Rehab Med. 2012;22(4):59-77.
  26. Kim KB, Ahn SH. The Efficacy of Ephedra sinica, Panax ginseng, and Alisma orientale Extract on Insulin resistance induced by Non-alcoholic fatty liver disease (NAFLD). J Pediatr Korean Med. November, 2020;34(4):11-21.
  27. Ki YB, Kim DH, Kang DH, Kim SJ, Choi JB, Yang MS. Effect of Mixed Extract of Ephedrae Sinica and Fibrosum Gypsum on Differentiation of Preadipocytes and Obesity of Rats. Journal of Korean Medicine Rehabilitation. 2014;24(3):11-27.
  28. Kim KS, Kim YB. A Case of Idiopathic Cholinergic Urticaria Associated with Acquired Hypohidrosis by Herbal Medicine Applied Gejimahwanggakbantang. The Journal of Korean Oriental Medical Ophthalmology & Otolaryngology & Dermatology 2009;22(3):220-7.
  29. Bae JY, Jo EH, Kim HY, Park MC. Inhibitory Effect of Ten Kinds of Herbal Medicine Including Ephedrae Herba on Nitric Oxide in LPS-induced Murine Peritoneal Macrophage. The Journal of Korean Oriental Medical Ophthalmology & Otolaryngology & Dermatology 2009;22(2):19-38.
  30. Kim JR, Yun SJ, Lee YK, Lee HJ, Kim JS. A Review on Safety of Herbal Medicines for Doping. J Korean Med. 2019;40(3):139-76.
  31. Jo GW, Ok JM, Kim SY, Lim YW. Review on the Efficacy and Safety of Mahuang and Ephedrine in the Treatment of Obesity -Focused on RCT-. J Korean Med. 2017;38(3):170-84.
  32. Chang MS, Shim KJ, Oh JH, Yoon JW, Choi MJ, Kim DR, Yang WM, Lim HH, Lee MJ, Park SK. Acute Oral Toxicity of Ephedrae Herba in SD Rats. Korean J. Oriental Physiology & Pathology. 2008;22(4):762-5.
  33. Choi DG, Shim KJ, Choi BJ, Park SY, Chang MS, Park SK. Subacute Oral Toxicity of Ephedrae Herba Extract in SD Rats. Kor. J. Herbology 2008;23(4):1-7.
  34. Ren Y, Chen Y, Tang EH, Hu Y, Niu B, Liang H, Xi C, Zhao F, Cao Z. Arbidol attenuates liver fibrosis and activation of hepatic stellate cells by blocking TGF-beta1 signaling. Eur J Pharmacol. 2024;967:176367.
  35. Dewidar B, Meyer C, Dooley S, Meindl-Beinker N. TGF-in Hepatic Stellate Cell Activation and Liver Fibrogenesis-Updated 2019. Cells. 2019;8(11):1419.
  36. Chen G, Xia B, Fu Q, Huang X, Wang F, Chen Z, Lv Y. Matrix Mechanics as Regulatory Factors and Therapeutic Targets in Hepatic Fibrosis. Int J Biol Sci. 2019;15(12):2509-21.
  37. Kim WH, Choi DY. Jangbubyeonjeungnonchi <7th Ed>. Seoul:Seongbosa. 2004:153-66.
  38. Kim JK, Han NR, Park SM, Jegal KH, Jung JY, Jung EH, Kim EO, Kim D, Jung DH, Lee JR, Park CA, Ku SK, Cho IJ, Kim SC. Hemistepsin A alleviates liver fibrosis by inducing apoptosis of activated hepatic stellate cells via inhibition of nuclear factor-κB and Akt. Food Chem Toxicol. 2020;135:111044.
  39. Chung JY, Chan MK, Li JS, Chan AS, Tang PC, Leung K, To K, Lan H, Tang PM. TGF-β Signaling: From Tissue Fibrosis to Tumor Microenvironment. Int J Mol Sci. 2021;22(14):7575.
  40. Kutz SM, Hordines J, McKeown-Longo PJ, Higgins PJ. TGF-β1-induced PAI-1 gene expression requires MEK activity and cell-tosubstrate adhesion. J Cell Sci. 2001;114:3905-14.
  41. Allen RR, Higgins PJ. Plasminogen activator inhibitor type-1 expression and the pathophysiology of TGF-β1-induced epithelial-tomesenchymal transition. Recent Res Devel Physiol. 2004;2:355-66.
  42. Desmouliere A, Geiniz A, Gabbiani F, Gabbiani G. Transforming growth factor-β 1 induces α-Smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol. 1993;122:103-11.
  43. Geervliet E, Bansal R. Matrix Metalloproteinases as Potential Biomarkers and Therapeutic Targets in Liver Diseases. Cells. 2020;9(5):1212.
  44. Ries C, Egea V, Karow M, Kolb H, Jochum M, Neth P. MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines. Blood. 2007;109(9):4055-63.
  45. Asrani SK, Devarbhavi H, Eaton J, Kamath PS. Burden of liver diseases in the world. J Hepatol. 2019;70:151-71.
  46. Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005;115:209-18.
  47. Fabregat I, Caballero-Diaz D. Transforming Growth Factor-β-Induced Cell Plasticity in Liver Fibrosis and Hepatocarcinogenesis. Front Oncol. 2018;8:357.
  48. Pellicoro A, Ramachandran P, Iredale JP, Fallowfield JA. Liver fibrosis and repair: immune regulation of wound healing in a solid organ. Nat Rev Immunol. 2014;14(3):181-94.
  49. Puche JE, Saiman Y, Friedman SL. Hepatic stellate cells and liver fibrosis. Compr Physiol. 2013;3:1473-92.
  50. Huang SJ, Chen SQ, Lin Y, Yang HY, Ran J, Yan FF, Huang M, Liu XL, Hong LC, Zhang XD. Maternal nicotine exposure aggravates metabolic associated fatty liver disease via PI3K/Akt signaling in adult offspring mice. Liver Int. 2021;41(8):1867-78.
  51. Dooley S, Delvoux B, Streckert M, Bonzel L, Stopa M, ten Dijke P, Gressner AM. Transforming growth factor beta signal transduction in hepatic stellate cells via Smad2/3 phosphorylation, a pathway that is abrogated during in vitro progression to myofibroblasts. TGF beta signal transduction during transdifferentiation of hepatic stellate cells. FEBS Lett. 2001;502:4-10.
  52. Ghosh AK, Vaughan DE. PAI-1 in tissue fibrosis. J Cell Physiol. 2012; 227:493-507.
  53. Iwaki T, Urano T, Umemura K. PAI-1, progress in understanding the clinical problem and its aetiology. British Journal of Haematology. 2012;157:291-8.
  54. Cho IJ, Kim SH, Kim SG. Inhibition of TGFb1-mediated PAI-1 induction by oltipraz through selective interruption of Smad 3 activation. Cytokine. 2006;35:284-94.
  55. Boo YC. Therapeutic Potential and Mechanisms of Rosmarinic Acid and the Extracts of Lamiaceae Plants for the Treatment of Fibrosis of Various Organs. Antioxidants (Basel). 2024;13(2):146.
  56. Lindert S, Wickert L, Sawitza I, Wiercinska E, Gressner AM, Dooley S, Breitkopf K. Transdifferentiation-dependent expression of alpha-SMA in hepatic stellate cells does not involve TGF-beta pathways leading to coinduction of collagen type I and thrombospondin-2. Matrix Biol. 2005;24:198-207.
  57. Rong X, Yang Y, Zhang G, Zhang H, Li C, Wang Y. Antler stem cells as a novel stem cell source for reducing liver fibrosis. Cell Tissue Res. 2020;379(1):195-206.
  58. Duarte S, Baber J, Fujii T, Coito AJ. Matrix metalloproteinases in liver injury, repair and fibrosis. Matrix Biol. 2015;44-46:147-56.