| Forest is an important resource for the production of natural products, and utilization of secondary metabolites of tree fruit or leaves to exploit drug, health food and so on, is an effective way to expand the medicine source, protect and use sustainably of resources. Catalpa bungei C. A. Mey. originated from China, which belongs to the Catalpa of Bignoniaceae family and is a deciduous tree, is the precious wood and famous ornamental tree species. In the situation of the forest property rights system reform and forest classification management, how to improve the economic benefits of non wood parts of C. bungei C. A. Mey. and achieve sustainable utilization of resources of C. bungei C. A. Mey. is becoming an urgent problem to be solved. This study mainly used leaves and seeds of the C. bungei C. A. Mey. as the materials and studied their bioactive components. Moreover, the effects of compound on He La cell proliferation and apoptosis were further explored. The main results were as follows.The amino acid, polypeptide, protein, sugar, glycosides, steroids, terpenoids, three phenols, flavonoids, lactones, coumarins, cardiac glycosides and alkaloids were determined according to the specific color and precipitation reaction of various chemical constituents through preliminary systematic test. The results also showed that chemical constituents of C. bungei C. A. Mey. Leaves probably contained tannins, anthraquinones but saponins, organic acids, volatile oil and fat composition had excluded.The Catalpa plant with high antioxidant activity was screened. The antioxidant activity and content of phenolic extracted from leaves of 12 clones Catalpa tree derived from Gansu and Henan provinces were compared. The results showed that total phenolic content and antioxidant activity in extract of C. bungei C. A. Mey. 6 leaves derived from Gansu province is the highest. Further comparing Catalpa fargesii Bur. with C. ovata G. Don., the total phenolic content and antioxidant activity in extract of CA6 leaves were significantly higher than C. fargesii Bur. and C. ovata G. Don..The antioxidative components were isolated from C. bungei C. A. Mey. leaves. The compounds were isolated by bioassay and column chromatography separation. The antioxidant activities of each separated fractions and compounds were evaluated. The results showed that the total phenolic content and antioxidant activities of ethyl acetate(EA) fraction in ethanol crude extracts were the highest, followed by n-butanol, petroleum ether(PE), and water fractions. Besides, 7 fractions were obtained from the EA fraction, in which the DPPH radical scavenging activity of Fr.5 was 2.86 times higher than BHT at the same concentration. Two flavonoid compounds with antioxidant activity were further isolated from the Fr.5 and they were luteolin(1) and apigenin(2), respectively, which were first founded in the leaves of C. bungei C. A. Mey..The antibacterial constituents were exploited based on the bioassay, column chromatography separation and size of the bacteriostatic ring of bacillus subtilis and bacillus cereus. The results showed that the antibacterial activity of ethyl acetate(EA) fraction was stronger than n-butanol, petroleum ether(PE), and water fractions. 29 parts were further obtained by separating the ethyl acetate(EA) fraction, in which the 19th(Fr.4.1) part showed higher antibacterial activity. The luteolin and apigenin were further isolated from the 19 th part. Each of two compounds showed poor antibacterial activity when they were separated. But combination of luteolin and apigenin demonstrated strong synergistic interaction. The appropriate ratio for two compounds was 1:1.The antitumor components were also isolated from C. bungei C. A. Mey. leaves. Using bioassay method and column chromatography separation combining with the method of SRB screened antitumor constituents inhibited He La cells proliferation from the leaves of C. bungei C. A. Mey.. The results showed that the ethyl acetate(EA) fraction had the strongest inhibitory effect to the He La cells proliferation compared to petroleum ether(PE), n-butanol, and water fractions. The 29 components isolated from ethyl acetate(EA) fraction all had the inhibitory effect on the He La cells proliferation. Among them, the 17th(Fr.3.11) showed the best inhibitory effect. The Fr.3.11 was further separated and then ursolic acid with antitumor activity was screened, which was first isolated from the leaves of C. bungei C. A. Mey..The effects of ursolic acid on the He La cells proliferation and apoptosis was investigated. Using the method of SRB, clone formation, scratch test and flow cytometry observed the influence of ursolic acid on the He La cells proliferation and cell cycle distribution. Using the Western Blotting detected the expression of protein during apoptosis process. The results indicated that the ursolic acid significantly inhibited He La cells proliferation and induce Hela cells apoptosis by entering the intrinsic and extrinsic pathway.The active ingredients of seed oil and defatted meal of C. bungei C. A. Mey. were analyzed. Using Gas chromatography-mass spectrometry(GC-MS) investigated constitutents of fatty acid and phytosterol of seed oil of C. bungei C. A. Mey.. Using high performance liquid chromatography(HPLC) detected the content of tocopherol of seed oil. Adopted high performance liquid chromatography-tandem mass spectrometry(HPLC-MS) investigated components and antioxidant activities of flavonoids in defatted meal. The results showed that 11 kinds of fatty acids have been detected, in which linoleic acid, α-linolenic acid and oleic acid were main components. The content of unsaturated fatty acids accounted for 87.37 % of fatty acids. Seed oil mainly contained campesterol(21.52 %), stigmasterol(4.43 %), β-sitosterol(74.05 %) and tocopherol(1.33 mg·g-1), in which the content of γ-tocopherol(0.89 mg·g-1) was about 2 times of α- tocopherol(0.44 mg·g-1). The defatted meal contained 7 kinds of flavonoids, and the content of total flavonoids was up to 9.39 mg·g-1. The extract of defatted seed meal also showed the antioxidant activity.Above results showed the active components and construction of the C. bungei C. A. Mey. seeds and leaves and explained the molecular mechanism induced apoptosis of tumor cells by active components. This study provided a theoretical basis and scientific guide for producing medicine and health product, developing new type of soil and utilizing comprehensively of C. bungei C. A. Mey.. |
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