Anti-inflammatory Efficacy and Liver Protective Activity of Pine Pollen according to Probe Sonicator Ultrasonic Disintegration Extraction Method

송화분의 초음파 파쇄 추출 방법에 따른 항염증 효능 및 간 보호 활성

  • Kim, Ok Ju (Department of Natural Science Institute, Silla University) ;
  • Woo, Young Min (Department of Natural Science Institute, Silla University) ;
  • Jo, Eun Sol (Department of Natural Science Institute, Silla University) ;
  • Jo, Min Young (Department of Pharmaceutical Engineering, Silla University) ;
  • Li, Chun-Ri (College of Acupuncture and Moxibustion, Liaoning University of Traditional Chinese Medicine) ;
  • Lee, Young-Ho (Protein Structure Research Group, Division of Bioconvergence Analysis, Korea Basic Science Institute, Bio-Analytical Science, University of Science and Technology (UST)) ;
  • Ahn, Mee Young (Department of Pharmaceutical Engineering, Silla University) ;
  • Lee, Sang-Hyeon (Department of Pharmaceutical Engineering, Silla University) ;
  • Ha, Jong Myung (Department of Pharmaceutical Engineering, Silla University) ;
  • Kim, Andre (Department of Pharmaceutical Engineering, Silla University)
  • 김옥주 (신라대학교 자연과학연구소) ;
  • 우영민 (신라대학교 자연과학연구소) ;
  • 조은솔 (신라대학교 자연과학연구소) ;
  • 조민영 (신라대학교 제약공학전공) ;
  • 이춘일 (중국 센양 요녕중의대) ;
  • 이영호 (한국기초과학연구소, 과학기술연합대학원) ;
  • 안미영 (신라대학교 제약공학전공) ;
  • 이상현 (신라대학교 제약공학전공) ;
  • 하종명 (신라대학교 제약공학전공) ;
  • 김안드레 (신라대학교 제약공학전공)
  • Received : 2019.07.09
  • Accepted : 2019.08.13
  • Published : 2019.10.10


In this study, the effect anti-oxidant, anti-inflammatory, and liver protective activity was investigated via quick ultrasonic disintegration of pine pollen using a probe sonicator (PS) followed by the extraction with water, 70% ethanol, and 100% ethanol. The anti-inflammatory effect was studied by measuring the production of nitric oxide (NO) and cytokine in RAW264.7 cells induced with lipopolysaccharides (LPS). The cell toxicity was also checked with an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the experiment was conducted using non-toxic $100{\mu}g/mL$. The NO inhibition rate was highest in the 70% ethanol PS group at $85.99{\pm}0.12%$. Also an excellent efficiency was obtained from the results of interlukin-1 beta ($IL-1{\beta}$) and tumor necrosis factor alpha ($TNF-{\alpha}$), which is related to inflammation-related cytokine, with the respective inhibition rates of 63 and 22%. To examine liver protective activity, HepG2 cells were treated with Taclin, and the generation of glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH) was measured in the culture solution. From GOT and LDH generation results, the inhibition rates in the 70% ethanol PS group were 28% and 13%, respectively, which was higher compared to that of using negative control group. Our results suggest that pine pollen extracted in 70% ethanol using PS may be used to develop food products that have anti-aging, anti-inflammatory, and liver protective effects.


Supported by : 중소벤처기업부


  1. B. Halliwell, Reactive oxygen species and the central nervous system, J. Neurochem., 59, 1609-1623 (1992).
  2. G. Block and L. Langseth, Antioxidant vitamins and disease prevention, Food Technol., 48, 80-85 (1994).
  3. H. L. Ko, K. H. Jegal, S. Y. Song, N. E. Kim, J. Kang, S. H. Byun, Y. W. Kim, I. J. Cho, and S. C. Kim, Water extract of Rosa leavigata Michx. protects hepatocytes from arachidonic acid and iron-mediated oxidative stress, Korean J. Herbology, 30, 7-15 (2015).
  4. F. P. de Heredia, S. Gomez-Martinez, and A. Marcos, Chronic and degenerative diseases obesity, inflammation and the immune system, Proc. Nutr. Soc., 71, 332-338 (2012).
  5. C. Nathan, Nitric oxide as a secretory product of mammalian cells, FASEB J., 6, 3051-3064 (1992).
  6. S. M. An, H. G. Kim, E. J Choi, H. H. Hwang, E. O. Lee, J. H. Baek, Y. C. Boo, and J. S. Koh, Screening for anti-inflammatory activities in extracts from Korean herb medicines, J. Soc. Cosmet. Sci. Korea, 40, 95-108 (2014).
  7. D. S. Lee, G. S. Jeong, R. B. An, B. Li, E. Byun, K. H. Yoon, and Y. C. Kim, Hepatoprotective effects of plants extracts from baekdu mountain on tacrine-induced cytotoxicity in HepG2 cells, Korean J. Pharmacogn., 39, 68-73 (2008).
  8. E. J. Cho and S. H. Yoon, Protective effect of asiasari radix on rat liver, J. Korean Soc. Hyg. Sci., 5, 85-91 (1999).
  9. P. B. Watkins, H. J. Zimmerman, M. J. Knapp, S. I. Gracon, and K. W. Lewis, Hepatotoxic effects of tacrine administration in patients with alzheimer's disease, JAMA, 271, 992-998 (1994).
  10. R. A. Osseni, C. Debbasch, M. O. Christen, P. Rat, and J. M. Warnet, Tacrine-induced reactive oxygen species in a human liver cell line: The role of anethole dithiolethione as a scavenger, Toxicol. In Vitro, 13, 683-688 (1999).
  11. M. L. Kim, Sensory characteristics of korean wheat noodles with pine pollen and antioxidant activities of pine pollen Extracts, Korean J. Food Cookery Sci., 21, 717-724 (2005).
  12. M. Gilliam, W. F. McCaughey, and B. Wintermute, Amino acids in pollens and nectars of citrus cultivars and in stored pollen and honey from honeybee colonies in citrus groves, J. Apic. Res., 19, 64-72 (1980).
  13. A. E. Vivino, and L. S. Palmer, The chemical composition nutritional value of pollen collected by bees, Arch. Biochem., 4, 129-136 (1994).
  14. B. Y. Lee, H. D. Choi, and J. B. Hwang, Components analysis of Korean pollens and pollen extracts, Korean J. Food Sci. Technol., 5, 869-875 (1997).
  15. M. A. Cribellaro, G. Senna, G. Riva, C. Cislaghi, P. Falagiani G. W. Canonica, and G. Passalacqua, Pollen mixtures used as health food may be a harmful source of allergens, J. Investig. Allergol. Clin. Immunol., 10, 310-311 (2000).
  16. B. Y. Lee, H. S. Chun, H, G. Kim, and H. M. Suk, Processing method of pollen, Korean Patent 10, 0211287, 0000 (1999).
  17. S. J. Kim, K. S. Youn, and H. S. Park, Antioxidative effect of pine, oak, and lily pollen exteracts, Korean J. Food Sci. Technol., 37, 833-837 (2005).
  18. M. S. Blois, Antioxidant determinations by the use of a stable free radical, Nature, 181, 1191-1200 (1958).
  19. S. Marklund and G. Marklund, Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase, Eur. J. Biochem., 47, 469-474 (1974).
  20. L. Prosky, N. G. Asp, T. F. Schweizer, J. W. Devries, and I. Furda, Determination of insoluble, soluble, and total dietary fiber in foods and food products: Interlaboratory study, J. Assoc. Off. Anal. Chem., 71, 1017-1025 (1988).
  21. J. M. McCord and I. Fridovich, Superoxide dismutase an enzymic function for erythrocuprein (hemocuprein), J. Biol. Chem., 244, 6049-6055 (1969).
  22. K. H. Lee, A. J. Kim, and E. M. Choi, Antioxidant and antiinflammatory activity of pine pollen extract in vitro, Phytother. Res., 23, 41-48 (2009).
  23. V. Goossens, J. Grooten, K. De Vos, and W. Fiers, Direct evidence for tumor necrosis factor-induced mitochondrial reactive oxygen intermediates and their involvement in cytotoxicity, Proc. Natl. Acad. Sci. USA, 92, 8115-8119 (1995).
  24. Y. K. Kim, Y. H. Kim, D. H. Kim, and K. T. Lee, Cytoprotective effects of natural flavonoids on carbon tetrachloride-induced toxiciy in prinary cultures of rat hepatocytes, Korean J. Pharmacogn., 36, 224-228 (2005).
  25. H. J. Kim, I. S. Lee, K. R. Lee, Antimutagenic and anticancer effects of Ramaria botrytis(Fr.) rick exstracts, J. Korean Soc. Food Sci. Nutr., 28, 1321-1325 (1999).