[논문]동충하초(Cordyceps militaris) 균사체 액체발효 포스트바이오틱스로부터 항염증 활성다당 분리

김연숙; 신현영; 김훈; 정은진; 김현경; 서민근; 서형주; 유광원

doi:10.9799/ksfan.2023.36.1.006

동충하초(Cordyceps militaris) 균사체 액체발효 포스트바이오틱스로부터 항염증 활성다당 분리
Anti-Inflammatory Active Polysaccharide from Postbiotics of Cordyceps militaris Mycelium-Liquid Culture 원문보기

한국식품영양학회지 = The Korean journal of food and nutrition, v.36 no.1, 2023년, pp.6 - 16

김연숙 (한국교통대학교 식품영양학전공) , 신현영 (고려대학교 대학원 의생명융합과학과 러닝헬스시스템융합전공) , 김훈 (중앙대학교 생명공학대학) , 정은진 (고려대학교 대학원 의생명융합과학과 러닝헬스시스템융합전공) , 김현경 (숙명여자대학교 식품영양학과) , 서민근 (고려대학교 대학원 의생명융합과학과 러닝헬스시스템융합전공) , 서형주 (고려대학교 대학원 의생명융합과학과 러닝헬스시스템융합전공) , 유광원 (한국교통대학교 식품영양학전공)

Abstract ▼ AI-Helper

To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

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표/그림 (7)

그림 Fig. 1. Fractionation of liquid culture by Cordyceps militaris mycelium (A) and molecular weight chromatogram of CM-CP and DCP (B).
$Table 1. HPLC analytical condition for the component sugar and molecular weight of subfraction from liquid culture by Cordyceps militaris mycelium$ 표 Table 1. HPLC analytical condition for the component sugar and molecular weight of subfraction from liquid culture by Cordyceps militaris mycelium
$Fig. 2. Cytotoxicity of subfraction from liquid culture of Cordyceps militaris mycelium against RAW 264.7 macrophage (A) and HaCaT keratinocyte (B). After each subfraction treated RAW 264.7 or HaCaT cells, inflammatory substances, such as lipopolysaccharide (LPS) or TNF-α+IFN-γ (T+I) was subsequently stimulated for 24 hr. A dexamethasone (50 and 20 μg/mL, for RAW 264.7 and HaCaT, respectively) was used as positive controls (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent tests in triplicate. Asterisks mean significant difference between inflammation group (LPS or T+I) and each group by Student's t-test. *p<0.05, **p<0.01, ***p<0.001.$ 그림 Fig. 2. Cytotoxicity of subfraction from liquid culture of Cordyceps militaris mycelium against RAW 264.7 macrophage (A) and HaCaT keratinocyte (B). After each subfraction treated RAW 264.7 or HaCaT cells, inflammatory substances, such as lipopolysaccharide (LPS) or TNF-α+IFN-γ (T+I) was subsequently stimulated for 24 hr. A dexamethasone (50 and 20 μg/mL, for RAW 264.7 and HaCaT, respectively) was used as positive controls (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent tests in triplicate. Asterisks mean significant difference between inflammation group (LPS or T+I) and each group by Student's t-test. *p<0.05, **p<0.01, ***p<0.001.
$Fig. 3. Anti-inflammatory activity of subfraction from liquid culture of Cordyceps militaris mycelium on LPS-stimulated RAW 264.7 cell. RAW 264.7 cells were treated with subfraction followed by lipolysaccharide (LPS) treatment for 24 hr, and production of IL-6 (A) or MCP-1 (B) was evaluated. A dexamethasone (50 μg/mL) was used as a positive control (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent tests in triplicate. Asterisks mean significant difference between the LPS group and each group by Student’s t-test. *p<0.05, **p<0.01, ***p<0.001.$ 그림 Fig. 3. Anti-inflammatory activity of subfraction from liquid culture of Cordyceps militaris mycelium on LPS-stimulated RAW 264.7 cell. RAW 264.7 cells were treated with subfraction followed by lipolysaccharide (LPS) treatment for 24 hr, and production of IL-6 (A) or MCP-1 (B) was evaluated. A dexamethasone (50 μg/mL) was used as a positive control (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent tests in triplicate. Asterisks mean significant difference between the LPS group and each group by Student’s t-test. *p<0.05, **p<0.01, ***p<0.001.
$Fig. 4. Anti-inflammatory activity of subfraction from liquid culture of Cordyceps militaris mycelium on TNF-α+IFN-γ- stimulated HaCaT keratinocyte. HaCaT keratinocyte were treated with each subfraction followed by TNF-α+IFN-γ (T+I) treatment for 24 hr, and the production of IL-6 (A) or IL-8 (B) was evaluated. A dexamethasone (20 μg/mL) were used as positive control (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent test in triplicate. Asterisks mean significant difference between T+I group and each group by Student's t-test. *p<0.05, **p<0.01, ***p<0.001.$ 그림 Fig. 4. Anti-inflammatory activity of subfraction from liquid culture of Cordyceps militaris mycelium on TNF-α+IFN-γ- stimulated HaCaT keratinocyte. HaCaT keratinocyte were treated with each subfraction followed by TNF-α+IFN-γ (T+I) treatment for 24 hr, and the production of IL-6 (A) or IL-8 (B) was evaluated. A dexamethasone (20 μg/mL) were used as positive control (PC). CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP. Results are expressed as mean±S.D. of three independent test in triplicate. Asterisks mean significant difference between T+I group and each group by Student's t-test. *p<0.05, **p<0.01, ***p<0.001.
$Table 2. Chemical property of subfraction from liquid culture by Cordyceps militaris mycelium$ 표 Table 2. Chemical property of subfraction from liquid culture by Cordyceps militaris mycelium
$Fig. 5. β-1,3/1,6-glucan content of subfraction from liquid culture of Cordyceps militaris mycelium. All results are expressed as mean±S.D. for three independent experiments. Different letters within a column are significantly different (p<0.05) by Duncan's multiple range test. CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP.$ 그림 Fig. 5. β-1,3/1,6-glucan content of subfraction from liquid culture of Cordyceps militaris mycelium. All results are expressed as mean±S.D. for three independent experiments. Different letters within a column are significantly different (p<0.05) by Duncan's multiple range test. CM-HW; postbiotics from C. militaris mycelium liquid culture, CP; crude polysaccharide of HW, and DCP; dialyzed-CP.

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저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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