Studies On The Chemical Constituents And Pharmacological Activities Of Platycladi Cacumen

Platycladi Cacumen is a traditional Chinese medicine with more kind medicinal value which recorded various traditional functions by many Chinese ancient herb books.In order to further clarify the promotes hair growth bioactive material basis of the lipid soluble constituents of the Platycladi Cacumen,this study conducted a detailed extraction and separation of the lipid soluble constituents from Platycladi Cacumen,and analyzed the volatile components.In addition,rosin paraffin wax mouse hair removal model was applied to study the pharmacologic activity of hair growth.In order to study its antitumor action of Platycladi Cacumen polysaccharides,the ultrasonic extraction process of polysaccharide from Platycladi Cacumen was optimized.The antitumor activity of polysaccharide in the Platycladi Cacumen was studied in order to provide scientific basis for the deep development and clinical application of Platycladi Cacumen.The research contents are as follows:A large number of literature showed that there were flavonoids,volatile oils,tannins,inorganic elements,etc chemical constituents in Platycladi Cacumen.There are many studies on the extraction,separation and content determination of flavonoids and volatile oils,but the development and utilization of the chemical components of Platycladi Cacumen are incomplete.Platycladi Cacumen has some pharmacological actions such as promoting hair growth,hemostasis,antibacterial,antioxidation and anti-tumor,but the potential pharmacological effects has not been fully developed.The pharmacological basis of the Platycladi Cacumen is rich,but the research on the chemical composition and pharmacological activity of the Platycladi Cacumen is only concentrated in the flavonoids and volatile oil,so the further exploration of other chemical constituents in Platycladi Cacumen has an extremely broad development prospect.In the chemical composition,two compounds Isopimara-7,15-dien-3β-ol（compound 1）and pinusolide（compound 2）were identified by the extraction,separation and purification of small molecules of lipophilic molecules in Plat ycladi Cacumen by the combination of the positive phase silica gel column chromatogra phy and the gel Sephadex LH-20 column chromatography.Isopimara-7,15-dien-3β-ol was extracted from Platycladi Cacumen for the first time.Forty-two compounds of five typie s alkenes,alcohols,ketones,aldehydes and esters were identified by GC-MS of Platycla di Cacumen lipid soluble constituents.The most content of the compounds was alkenes.After analysis and comparison,the two monomer compounds identified were not detected in GC-MS,the two compounds identified by the separation were not found in the result s of GC-MS detection.The two compounds both of which were characterized by low p olarity,high fat-soluble and non-volatile.At the same time,the residues from Platycladi Cacumen were removed and the polysaccharides from Platycladi Cacumen were obtaine d after removing the lipid soluble constituents.In order to establish mathematical models and to obtain optimal process,extraction temperature,extraction time,and liquid-solid rat io were selected as the independent variables,and the yield of polysaccharides was used as response value based on single-factor experiments.The optimal extraction conditions were obtained as follows:optimal extraction temperature 72℃,extracting time 67 min,li quid-solid ratio 19:1,experimental extraction yield 6.87±0.18%。In the field of pharmacological activity（1）Study on antitumor activity of crude polysaccharide from Platycladi Cacumen.The spleen index and thymus index of the tumor suppressor rate were calculated and the level of cytokines in serum and the antioxidant capacity of the body were detected by establishing the model of H₂₂ mice subcutaneous transplantation tumor.Furthermore,the antitumor activity and mechanism of the polysaccharides of Platycladi Cacumen were preliminarily discussed according to the H&E and Hoechst33258 two staining methods.The tumor inhibition rates of the low,medium and high three dose groups were 34.83%,42.70%and 49.44%.Moreover,the polysaccharide of Platycladi Cacumen protected the immune organs of the body,and adjust the content of IL-2,TNF-αand IFN-γin the serum of H₂₂ bearing mice to the normal level.The polysaccharide of Platycladi Cacumen reduced the antioxidant activity index MDA content and significantly increase the activity of CAT and SOD.Thus,the content of lipid peroxides is reduced,which has antioxidant effect,and finally the activity of anti-tumor is played.（2）Study on the active constituents of the Hair growth active ingredients in Platycladus orientalis.The pharmacological effects of Isopimara-7,15-dien-3β-ol（compound 1）and pinusolide（compound 2）of Platycladi Cacumen on hair growth were studied,and the mechanism was preliminarily discussed.The newborn hair length and weight,the level of cytokine VEGF and TNF-αwere detected by rosin-paraffin hair removal,and the morphology of newborn hair follicles of the hair was observed.The results showed that the effect of the pinusolide on the growth of newborn hair length was obvious（P<0.01）.Both Isopimara-7,15-dien-3β-ol and pinusolide significantly increased the weight of newborn hair（P<0.01）.Both Isopimara-7,15-dien-3β-ol and pinusolide were significantly increased the levels of VEGF and the levels of TNF-αwere significantly increased by pinusolide（P<0.05 or<0.01）.The results of tissue section observation showed that two compounds in the Platycladi Cacumen could improve the vigor of the hair follicle,maintain the normal growth state of the hair follicle,and promote the growth of hair.Conclusion:This study shows that the polysaccharide of the Platycladi Cacumen has antitumor activity which achieved by improving the immune function and anti-oxidative ability of the body.In addition Isopimara-7,15-dien-3β-ol and pinusolide have the effect of promoting hair growth.In this study,the active material bases of hair growth were identified Isopimara-7,15-dien-3β-ol and pinusolide.