Studies On The Anti-inflammatory Constituents Of Rhododendron Mariae Hance

Rhododendron amesiaeRehd.et Wils,also known as Rhododendron mariaeHance,belongs to genus Rhododendron by morphological and DNA analysis,which is widely distributed in Jiangxi,Hunan,Guangdong,Guangxi provinces,etc.It has been documented that the leaves and shoots of Rhododendron mariae contain volatile oils,various terpenoids,phenols,organic acids and tannins.It has also been reported that the rhododendron leaves contain coumarin,lactone,oxime and oleanolic acid.The main active constituents of Rhododendron mariaeare are quercetin,hyperoside,rhododendrin and other flavonoids,which have antitussive,expectorant,anti-inflammatory,antibacterial,smooth muscle spasm,respiratory inhibition,central inhibitioneffects,especially for treating asthma,phlegm and bronchitis.There are few literatures on Rhododendron mariae at home and abroad,and it is widely distributed in Jiangxi,Hunan,Guangdong,Guangxi and other areas with toxic homologous plants such as Yellow Azalea and A.rugulosa,soit is not clear that ifR.mariaecontains the xylosin-type diterpenoid toxic substances.Most of the existing papers believe that the anti-inflammatory,anti-inflammatory and antitussive activities of R.mariae are derived from the flavonoids that have been isolated and confirmed.In order to demonstrate the correctness of this view,and it is also to find out whether there are other structurally novel compounds in the R.mariae.It can provide a scientific basis for the secondary development of the two drug approvals of R.mariae tablets and zicha granules retained byGuangzhou Baiyun Mountain Hutchison Whampoa traditional Chinese Medicine Co.,Ltd.?Guangyao Group subsidiary company?.We decided to further study the chemical constituents and pharmacological effects of R.mariae.In this thesis,the chemical constituents of 70%ethanol extract of R.mariaewere studied,and the anti-inflammatory activityon zebrafish model were performed on total extract,each fraction and pure compounds obtained from the extraction of 70%alcohol extract and macroporous resin.1.The study on chemical components ofR.maria.In this thesis,11 compounds,including one flavonoid,three andrographolide and seven triterpenoids,were isolated and identified from 70%ethanol extract of R.mariae by various modern chromatographic separation methods and spectroscopic methods.They are3-dehydro-deoxyandrographolide?DJ-1?,deoxy-andrographolide?DJ-2?,dehydrated andrographis Lactone?DJ-3?,2?,3?,24?-Trihydroxyurs-12-en-28-oicacid?DJ-4?,2?,3?-dihydroxy-12-ursen-28-oic acid?DJ-5?,2?,3?-dihydroxy-12-oleanen-28-oic acid?DJ-6?,2?,3?,24?-trihydroxyolean-12en-28-oic acid?DJ-7?,2?,3?,24?-trihydroxy ursolic acid?DJ-8?,hypotatic acid-A?DJ-9?,2?,3?,24?-trihydroxy-12-en-urs-28-oic acid?DJ-10?,and farrerol?DJ-11?.Among them,the compoundsDJ-1,DJ-2,DJ-3 are andrographolidediterpenoids,which are isolated from family Ericaceaefor the first time.The triterpenes,DJ-4,DJ-7,DJ-9 and DJ-10 are isolated from genus Rhododendronfor the first time,while DJ-5,DJ-6andDJ-8were isolated fromR.mariaefor the first time.2.The preliminary study on the anti-inflammatory effects of R.mariaeThe CuSO₄ induced inflammation zebrafish model and Tail-cutting model were established,and preliminary study on anti-inflammation effects of total ethanol extract,macroporous resin fraction and the isolated compounds from R.mariae were carried out.The results showed that the total extract?concentration:100?g/ml?could significantly reduce the number of macrophages in the inflammatory model of zebrafish induced by copper sulfate and the number of macrophages aggregated in the broken tail of zebrafish.Compared with sub-fractions and monomer compounds,the total extract had the strongest anti-inflammatory activity,and had more significant anti-inflammatory effect than DEX group?concentration:5?g/ml?.The fractions showed different anti-inflammatory effects in two zebrafish models of acute inflammation.Among them,DJ1-12 and DJ13-16 fractions showed good anti-inflammatory activity in both anti-inflammatory models,second only to the total extract.Diterpenoids have good anti-inflammatory effects in both CuSO₄induced acute inflammation model and tail-amputated acute inflammation model of zebrafish.The anti-inflammatory activity of DJ-2?concentration:50?g/ml?was the strongest,which was second only to that of DEX?concentration:5?g/ml?.Triterpenoid monomers showed anti-inflammatory effects in both CuSO₄induced acute inflammation model and tail-amputated acute inflammation model in zebrafish.