Effects Of Angelica On Proliferation Of Murine Bone Marrow Mononuclear Cell And Its Cellular Signaling Pathways

The aim of this thesis is to understand regulating effects of several Traditional Chinese Medicine (TCM)herbs,especially Angelica Sinensis(Oliv.)Diels,on proliferation of murine bone marrow cell,and its intracellular signal pathways.The paper divided into three parts.The first part aimed at choice of herbs and the best working condition.ln this part we chose four typical herb,.Angelica for "enrich the Blood",Astragalus, for "regulate vital energy",Lycium Barbarum for "nourish Yin", and Tripterygium for "Expel wind-evil and remove wetness-evil" .Effects of these herbs on proliferation of murine bone marrow cells were investigated.Result of CFU-GM and MPO showed stimulating effects from extract of Angelica and Astragalus. Dosage and effect relationship were found at herb concentration of 0.1mg/ml,1mg/ml,and 10mg/ml.The maximal effect of Angeilca extract was observed at 10mg/ml with 73% increased CFU-GM count and 369% increased MPO enzyme/protein ratio.The maximal effect of Astragalus extract was found at 1mg/ml with 22% increased CFU-GM count and 96% increased MPO enzyme/protein ratio.Extract of Lycium Barbarum-LBP was also studied and no effect was detected at dose of 0.01mg/ml,0.1mg/ml,1mg/ml.Effect of Tripterygium preparation-TGW,is inhibited.The maximal effect was observed at concentration of 1mg/ml with 41% decreased CFU-GM count and 77% decreased MPO enzyme/protein ratio.According these results,we chose Angelica for further investigation.The next step is to chose the best working condition.Murine bone marrow mononuclear cell and MTT assay were used to make out the cell growth curve.We found day3 is suitable time for our study,and the best concentration of Angelica injection is lmg/ml.Culture medium was checked for the best effect.10%FBS-IMDM,5%FBS-IMDM and FBS-free-IMDM were tested.We found 444% increased cell growth at the dose of lmg/ml Angelica injection.In 10%FBS-IMDM and 5% FBS-IMDM,the background proliferation is strong which covered parts of effect of Angelica injection.In the second part we investigated the effect of Angelica on MAPK pathway.MAPK pathway is one of important ways in cell growth regulation.The key process is MAPKKK→MAPKK→MAPK.Erk1/2,SAPK/JNK and P38 are three main types of MAPKs with their special antibodies.Our experiments with Angelica added in mononuclear cell indicated that ERK1/2 and P38 MAPK were activated from 30 min after the herb preparation added,while SAPK/JNK MAPK keeps no change.Further investigated was did with MAPK inhibitor.Co-culture with PD98059,the ERK1/2 ihibitor,Angelica was found a 57.3% decreased effect on cell proliferation ,and with SB 203580,the P38 inhibitor,42.7% decreased effect was observed.So we proved that Angelica induced proliferation of murine boe marrow mononuclear cell was transmitted by ERK1/2 and P38 MAPK.The third part of this thesis discussed effect of Angelica on JAK-STAT pathway-another important pathway involving in cell growth regulation.