| Purpose:To study the chemical constituents of Pinus massoniana pollen in order to provide theoretical and experimental evidence for further exploitation of the medicinal value of Pinus massoniana pollen and the protective effects of Pinus massoniana pollen on experimental gastric mucosa in rats.Material and method:The compounds were isolated and purified by repeated column chromatography and their structures were identified on the basis of their chemical and physical properties as well as the spectra data of ESI-MS,1H-NMR, 13C-NMR etc, compared with data reported in literatures. On basis of the study chemical constituents from Pinus massoniana pollen, effect of Pinus massoniana pollen on gastric ulcers in rats were studied.Rats were divided randomly into six groups, normal control group, model control group, groups with tested drug (setting three groups which were administrated respectively Pinus massoniana pollen at the dose of 900,1800,3600mg/kg) and Cimitidine group. All rats were separately administered at 10ml/kg to each group for fourteen days. All rats were deprived of food but allowed free access to water for 24hr after the last administration, and then the model of gastric ulcer induced by ethyl alcohol and indomethacin were built, In the model induced by ethyl alcohol,1 hour after building the model, collecte blood, observe the index of the injury of gastric mucosa, and determine ulcer index. Furthermore, the contents of nitric oxide (NO), malonaldehyde (MDA) and the activities of superoxide dismutase (SOD) in blood serum were examined. The model of acetic acid-induced gstric ulcers were made as follows: under ether anesthesia, the abdomen was incised and stomachs exposed. A round plastic mold (5 mm in diameter) was placed in contact with the serosal surface of the corpus mucosa, and acetic acid 75μl was poured into the mold and left in place for 60 seconds. after the acid was removed, the treated surface was rinsed with saline and the abdomen was closed, and the animals were fed normally, on the first day after the operation, the rats were randomized into 5 groups:model control group, Cimitidine group, Pinus massoniana pollen groups (setting three groups which were administrated respectively Pinus massoniana pollen at the dose of 900,1800,3600mg/kg) groups. The rats of model group were adminisitered saline. All rats were seperately administered at 10ml/kg to each group every day for 14 continuous days. All rats were deprived of food but allowed free access to water for 24hr after the last administration, observe the index of the injury of gastric mucosa, and determine ulcer index.Results:19 compounds had been isolated from the extracts,16 compounds were identified as:(3-sitosterol (1),2,3-Dihydroxypropyl octadecanoate (2), hexadecan-oic acid 2,3-dihydroxy-propyl ester (3), daucosterol (4),1,3-Dimyristin (5), myristic acid (6), 14-Methylhexadecanoic acid (7), p-Hydroxybenzaldehyde (8),3-hydroxy-4-methoxy-benzoic acid (9), succinic acid (10),3,3',5,5',7-pentahydroflavanone (11), dihydrokaempferol (12), p-hydroxybenzcic acid (13),3,4- dihydroxybenzoic acid (14), kaempferol (15), uracil (16); The results of the study of protective effects of Pinus massoniana pollen on experimental gastric mucosa in rats showed that the middle and high dose groups of Pinus massoniana pollen could significantly inhibit the ulcer areas (p<0.05, p<0.01), Pinus massoniana pollen could inhibit elevation of MDA (P<0.05) and decrease of NO level (P<0.05), as well as the activities of SOD (p<0.05, p<0.01).Conclusion:Isolation and identification of 19 compounds,15 compounds were obtained from Pinus massoniana pollen for the first time:2,3-Dihydroxypropyl octadecanoate (2), hexadecan-oic acid 2,3-dihydroxy-propyl ester (3), daucosterol (4),1,3-Dimyristin (5), myristic acid (6),14-Methylhexadecanoic acid (7), p-Hydroxybenzaldehyde (8), 3-hydroxy-4-methoxy- benzoic acid (9), succinic acid (10),3,3',5,5',7-pentahydroflavanone (11), dihydrokaempferol (12), p-hydroxybenzcic acid (13),3,4- dihydroxybenzoic acid (14), kaempferol (15), uracil(16). The experimental results showed that Pinus massoniana pollen has protective effects against gastric mucosa injury, and the mechanism is probably related with the increasingly release of NO in plasm, enhencement of SOD activity, decrease of MDA level. |
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