Study On Active Substance And Quality Analysis Of Rhizoma Panax Japonicus

Rhizoma Panax japonicus is the dried rhizome of Panax japonicus, belonging to Panax species of Araliaceae.As a traditional Chinese medicine,it is widely used as a tonics.Meanwhile,it possesses many pharmacological effects,such as antiinflammation,immunoiogicai regulation,anti-tumor, protecting myocardial ischemia,activating blood circulation,detumescence and analgesia,etc.Clinically,it is commonly used in asthenia,cough and hemoptysis,rheumatism.This thesis focused on the screening of active fractions,observation of several pharmacological effects,elucidating mechanism of principal effects, establishment of qualitiy evaluation methods for crude drugs and optimization of producing procedure of crude active products,which lay a solid foundation for the further development of the titled medicine.Its anti-inflammatory actions were observed in vivo against mice ear oedama induced by dimethylbenzene and mice paw swelling induced by Freund's complete adjuvant.Results showed that TSPJ significantly inhibited dimethlybenzene-induced ear oedama and Freund's complete adjuvant-induced paw swelling in mice,and the inhibitory effect of TSPJ at a high dose was superior to that of total glucosides of paeony,i.e.TSPJ had significantly anti-inflammatory actions.The curative effects of TSPJ towards adjuvant arthritis and collagen-induced arthritis were observed in adjuvant arthritis rats induced by complete Freunds adjuvant in vivo and inhibitory action of rats induced by collagen on arthritis synovial cell in intro.Results showed that TSPJ decreased Freund's complete adjuvant-induced paw swelling and the levels of TNF-a and IL-1βin serum of AA rats,which was dose-related;in addition,the significantly inhibitory effects of the samples in the proliferation of synovial cells and the apoptosis rate of rats.The results above inferred that the anti-inflammatory actions of TSPJ be related to the regulation of the the level of TNF-a and IL-1β.The thesis involved in the observation of the effects of TSPJ on a solid tumor,an ascites tumor and proliferation and differentiation of human leukemic cells(HL-60) in vivo and in vitro on mice.The experiments showed that TSPJ markedly inhibited the growth of S-180 tumor,improved the life quality and prolonged the life-span of the mice.The results revealed that TSPJ markedly inhibited the growth of HL-60,and invoked differentiation of granular cells.The protection of TSPJ on the heart muscle ischemia of rats were studied. TSPJ significantly decreased the level of cytokine LDH,CK,MDA in the serum of rats,and promoted the superoxide dismutase in MI rats.It was concluded that the effects above were paralleled with its capacities to remove active oxygen free radicals and inhibit lipid peroxidation.Based on pharmacological activity screening,two extraction methods were adopted to separate and purify total saponins of Panax japonicus,i.e. traditional method and foam separation.The latter was a newly established procedure to separate saponins and related parameters were optimized.The chemical constituents of the titled plant were obtained by column chromatography and HPLC over macroporous adsorptive resins and silica gel. Three compounds were obtained,and two of them were identified as ginsenoside Rd and ginsenoside Re,respectively.According to reports and our experimental results,ginsenosides accounted for most of the activities of Panax japonicus.In order to evaluate the quality of crude drugs,a photometric method was established to determine the content of TSPJ in Rhizoma Panax japonicus with vanillin-perchloric acid as chromogenic agent and detection wavelength at 552 nm.The results showed that the contents of TSPJ were more than 16%in different batches of Panax japonicus.A HPLC method was used to determine the content of oleanolic acid with methanol:water:glacial acetic acid:triethylamine= 90:10:0.04:0.02 as the mobile phase.The results suggested that the content of free oleanolic acid was much less that of combined oleanolic acid,with the latter being up to 99%.Two HPLC methods,i.e.HPLC-UV and HPLC-ELSD,were established to obtain the HPLC fingerprint chromatograms of Panax japonicus. Experimental results showed that both methods exhibited high sensitivity and good reproducibility,which improved the quality standards of this medicine recorded on 2005 Edition of "People's Republic of China Pharmacopoeia".