Research On The Anti-inflammative Constituents From The Seeds Of Quercus Serrata Var. Brevipetiolata

Quercus serrata var. brevipetiolata is a variant belonging to Fagaceae, Quercus genus. According to the “Flora of China”, there are about 300 species of Quercus genus around the world, in which China has 51 species, 14 varieties and 1 form. The plants are mainly distributed in southern Liaoning, Shanxi, Gansu, Shandong, Jiangsu, Anhui, Zhejiang, Jiangxi, Fujian, Taiwan, Henan, Hubei, Hunan, Guangdong, Guangxi, Sichuan, Guizhou, and et al, and many of them are economic plants both in traditional Chinese medicine and food.At present, there are many studies on the genus of Q., but fewer of the chemical components and pharmacological activities on the plant of Q. serrata var. brevipetiolata, and research about its seed has not been found yet. According to the relevant literatures, the main chemical compositions isolated from the genus are flavonoids, triterpenoids, tannins, alkaloids, volatile oil, and et al. And modern pharmacological researchs have showed that these compounds display good effects of antioxidant, antibacterial and antiviral, anti-inflammatory, anti-tumor. The purpose of this research was focused on the bioactive components of the seeds of Q.serrata var. brevipetiolata.In this study, 34.5 kg seeds of Q. serrata var. brevipetiolata were extracted four times with 70% ethanol under reflux. And the solvent was evaporated in vacuo to gain 30 L aqueous residue. The aqueous residue was partitioned successively with cyclohexane, Et OAc, and n-Bu OH. After preliminary analysis by TLC, the ethyl acetate extracts were futher isolated by means of silica gel column chromatography, Sephadex LH- 20, ODS and preparative HPLC, and 34 compounds were isolated and purified. Based on their chemicophysical properties and spectroscopic data, the structures of the 34 compounds were identified as 2?,3?,19?-Trihydroxy-24-oxo-olean-12-en-28-oic acid(1*), 3?-acetoxy-2?,19?,23-Trihydroxyurs-12-en-28-oic acid(2*), 3,23-OMethyl butyrate-2?,3?,19?,23-tetrahydroxy-olean-12-en-28-oic acid(3*), 3,23-O-Methyl butyrate-2?,3?,19?,23-tetrahydroxy-olean-12-en-28-oic acid ?-Dglucopyranoside ester(4*), 3,23-O- Methyl butyrate-2?,3?,19?,23-tetrahydroxy-urs-12-en-28-oic acid ?-D-glucopyranoside ester(5*), 3?,23-Dihydroxy-24-oxo-olean-13(18)-en-28-oic acid 28-O-?-D-glucopyranoside ester(6*), Oleanolic acid(7), Arjunolic acid(8), Arjunglucoside II(9), Arjungenin(10), Arjunglucoside I(11), Arjunic acid(12), 24-Deoxysericoside(13), Sericic acid(14), Sericoside(15), 2?,3?,19?-Trihydroxy-olean-12-en-24,28-dioic acid 28- β-D-glucopyranoside ester(16), Paradrymoniside(17), Glucosyl tormentate(18), 4-epi-Niga-ichigoside F1(19), 2?,3?,19?,23- Tetrahydroxy-urs-12-en-28-oic acid(20), Niga-ichigoside F1(21), Trachelosperoside A-1(22), Pinfaenoic acid(23), 2?,3?,23-Trihydroxy-olean-12,18(19)-dien-28-oic acid ?-D-glucopyranoside ester(24), 2?,3?,23-Trihydroxy-urs-12,19(20)-dien-28-oic acid(25),2?,3?,23-Trihydroxyurs-12,19(20)-dien-28-oic acid 28-O-?-D-glucopyranoside ester(26), 2?,3?,23-Trihydroxyurs-12,19(29)-dien-28-oic acid(27), Quadranoside VIII(28), Quadranoside X(29), 2?,3?,23-Trihydroxyurs-11,13(18)-dien- 28-O-?-D-glucopyranoside ester(30), 5-Hydroxymethylfurfural(31), Gallic acid(32), ?-Daucosterol(33), Palmitic acid(34), respectively. Compounds 1-6 were new, and all of the above were isolated for the first time from this plant. Meanwhile, all isolated compounds were tested for their inhibitory effects on LPSinduced nitric oxide production in RAW 264.7 macrophages. Except compounds 9, 15 and 16, others showed obvious effect of anti-inflammatory activities. Compounds 1, 4, 7, 11, 13, 23, 25, 27 exhibited stronger anti-inflammatory activities with IC50 values of 5.4, 8.2, 7.8, 4.0, 8.9, 8.6, 6.3 and 5.4 ?M, respectively, and compounds 2, 5, 8, 12, 14, 17, 18, 19 and 21 also showed moderate anti-inflammatory activities with IC50 values of 19.1, 12.8, 13.0, 20.1, 17.2, 10.1, 10.4, 10.3, 11.6, 12.3 ?M, compared with the positive control drug indomethacin(IC50 = 47.4 ?M). In addition, compounds 1, 2, 4, 5 were also tested for their inhibitory effects on TNF-? induced IL-6 and IL-8 production in MH7 A, and results showed that the four compounds can also exertanti-inflammatory effects by inhibiting the production of IL-6 and IL-8.