Anti-inflammatory activity of crude extracts from some traditional medicinal plants

Test Samples (12 종 )  금은화 ( 金銀花 ) Lonicera japonica (Caprifoliaceae)  두충 ( 杜食 ) Eucommia ulmoides (Eucommiaceae)  만형자 ( 蔓荊子 ) Vitex trifolia (Verbenaceae)  산수유 ( 山茱萸 ) Cornus officinalis (Cornaceae)  생지황 ( 生地黃 ) Rehmanniae glutinosa var. purpurea (Scrophulariaceae)  용담 ( 龍膽 ) Gentiana scabra (Gentianaceae)  인동 ( 忍冬 ) Lonicera japonica (Caprifoliaceae)  정향 ( 丁香 ) Eugenia caryophyllata (Myrtaceae)  차전자 ( 車前子 ) Plantago asiatica (Plantaginaceae)  치자 ( 梔子 ) Gardenia jasminoides (Rubiaceae)  패장 ( 敗醬 ) Patrinia villosa (Valerianaceae)  현삼 ( 玄蔘 ) Scrophularia ningpoensis (Scrophulariae)

Photos 금은화 두충만형자산수유 생지황용담 인동 정향 차전자치자 패장 현삼

Objective  약용식물 12 종의 항염작용 고찰  Cytotoxicity (MTT)  COX-1 activity (TXB2)  COX-2 activity (PEG2)  TNFα formation  NO production

Crude Extraction H2OH2O MeOH

Cytotoxicity (MTT) assay for extracts Principle MTT Formazan reduction by mitochondrial respiration Method  RAW cell culture and seeding in a 96-well plate  LPS and a crude extract (100μg/ml) treatment and incubation for 20hrs at 37 ℃  Incubation with MTT(0.1mg/ml) for 4hrs at 37 ℃  Adding DMSO (200μl) to dissolve formazan crystal  Absorbance measurement at 450 and 650nm using automatic ELISA microplate reader

Cytotoxicity (MTT) assay for extracts Result H2OH2O MeOH LPS(+) 100 μg/ml

Cytotoxicity (MTT) assay for extracts Result H2OH2O MeOH LPS(+) 100 μg/ml

Effects of extracts on TXB 2 production  HEL cell culture and seeding in a 96-well plate  A crude extract (1,10, 50, 100 ㎍ /ml) treatment and incubation for 1hr  Arachidonic acid (30 μM ) treatment for 15min  Preparation of cell lysate  ELISA using anti-TXB 2 monoclonal antibodies  IC 50 calculation Method

Effects of 금은화 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 금은화 Ex : 1/10/50/100 μg/ml

Effects of 두충 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 두충 Ex : 1/10/50/100 μg/ml

Effects of 만형자 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 만형자 Ex : 1/10/50/100 μg/ml

Effects of 산수유 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 산수유 Ex : 1/10/50/100 μg/ml

Effects of 생지황 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 생지황 Ex : 1/10/50/100 μg/ml

Effects of 용담 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 용담 Ex : 1/10/50/100 μg/ml

Effects of 인동 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 인동 Ex : 1/10/50/100 μg/ml

Effects of 정향 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 정향 Ex : 1/10/50/100 μg/ml

Effects of 차전자 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 차전자 Ex : 1/10/50/100 μg/ml

Effects of 치자 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 치자 Ex : 1/10/50/100 μg/ml

Effects of 패장 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01, 0.1, 1, 10 μM Aspirin : 1, 10, 50, 100 μM 패장 Ex : 1, 10, 50, 100 μg/ml

Effects of 현삼 Ex on TXB 2 production Result ELISA (n=3) Piroxicam : 0.01, 0.1, 1, 10 μM Aspirin : 1, 10, 50, 100 μM 현삼 Ex : 1, 10, 50, 100 μg/ml

Effects of extracts on PEG 2 production  RAW cell culture and seeding in a 96-well plate  A crude extract (1,10, 50, 100 ㎍ /ml) treatment and incubation for 1hr  Arachidonic acid (30 μM ) treatment for 15min  Stimulation with LPS(1 ㎍ /ml) and IFN-γ(10U/ml) for 4hr  Preparation of cell lysate  ELISA using anti-PGE 2 monoclonal antibodies  IC 50 calculation Method

Effects of 금은화 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 금은화 Ex : 1/10/50/100 μg/ml

Effects of 두충 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 두충 Ex : 1/10/50/100 μg/ml * * *

Effects of 만형자 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 만형자 Ex : 1/10/50/100 μg/ml

Effects of 산수유 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 산수유 Ex : 1/10/50/100 μg/ml

Effects of 생지황 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 생지황 Ex : 1/10/50/100 μg/ml

Effects of 용담 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 용담 Ex : 1/10/50/100 μg/ml

Effects of 인동 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 인동 Ex : 1/10/50/100 μg/ml

Effects of 정향 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 정향 Ex : 1/10/50/100 μg/ml

Effects of 차전자 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 차전자 Ex : 1/10/50/100 μg/ml * * *

Effects of 치자 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 치자 Ex : 1/10/50/100 μg/ml

Effects of 패장 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 패장 Ex : 1/10/50/100 μg/ml

Effects of 현삼 Ex on PEG 2 production Result ELISA (n=3) Meloxicam : 0.01/0.1/1/10 μM Aspirin : 1/10/50/100 μM 현삼 Ex : 1/10/50/100 μg/ml

Effects of extracts on TNF  production  RAW cell culture and seeding in a 96-well plate  A crude extract (1,10, 50, 100 ㎍ /ml) treatment and incubation for 1hr  Stimulation with LPS(1 ㎍ /ml) and IFN-γ(10U/ml) for 4hr  Preparation of cell lysate  ELISA using anti-TNF  monoclonal antibodies  IC 50 calculation Method

Effects of 금은화 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 금은화 Ex : 1/10/50/100 μg/ml

Effects of 두충 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 두충 Ex : 1/10/50/100 μg/ml

Effects of 만형자 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 만형자 Ex : 1/10/50/100 μg/ml

Effects of 산수유 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 산수유 Ex : 1/10/50/100 μg/ml

Effects of 생지황 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 생지황 Ex : 1/10/50/100 μg/ml

Effects of 용담 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 용담 Ex : 1/10/50/100 μg/ml

Effects of 인동 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 인동 Ex : 1/10/50/100 μg/ml

Effects of 정향 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 정향 Ex : 1/10/50/100 μg/ml

Effects of 차전자 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 차전자 Ex : 1/10/50/100 μg/ml * * *

Effects of 치자 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 치자 Ex : 1/10/50/100 μg/ml

Effects of 패장 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 패장 Ex : 1/10/50/100 μg/ml

Effects of 현삼 Ex on TNFα production Result ELISA (n=3) Rolipram : 0.01/0.1/1/10 μM 현삼 Ex : 1/10/50/100 μg/ml

Effects of extracts on NO production  RAW cell culture and seeding in a 96-well plate  A crude extract (1,10, 50, 100 ㎍ /ml) treatment and incubation for 1hr  Stimulation with LPS(1 ㎍ /ml) and IFN-γ(10U/ml) for 24hr  Preparation of cell lysate  The isolated supernatants were mixed with an equal volume of Griess reagent  Nitrite production was determined by measuring the absorbance at 550 nm  IC 50 calculation Method

Effects of 금은화 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 금은화 Ex : 1/10/50/100 μg/ml

Effects of 두충 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 두충 Ex : 1/10/50/100 μg/ml

Effects of 만형자 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 만형자 Ex : 1/10/50/100 μg/ml

Effects of 산수유 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 산수유 Ex : 1/10/50/100 μg/ml

Effects of 생지황 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 생지황 Ex : 1/10/50/100 μg/ml

Effects of 용담 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 용담 Ex : 1/10/50/100 μg/ml

Effects of 정향 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 정향 Ex : 1/10/50/100 μg/ml

Effects of 차전자 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 차전자 Ex : 1/10/50/100 μg/ml * *

Effects of 치자 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 치자 Ex : 1/10/50/100 μg/ml

Effects of 패장 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 패장 Ex : 1/10/50/100 μg/ml

Effects of 현삼 Ex on NO production Result ELISA (n=3) L -NAME : 1/10/50/100 μM 현삼 Ex : 1/10/50/100 μg/ml

IC 50 of aqueous extracts for COX-1, COX-2, TNFα and NO Result COX-1COX-2TNFαNO A- 차전자 A- 두충 IC 50 ( μg/ml )