| Objectives:To study the preventive and therapeutic effects of chlorophyll-carotene paste from pine needles on experiment gastric ulcer, in order to provide theoretical and experimental evidence for further exploitation of the medicinal value of the pine needles.Methods:(1) The safety of the active fraction was evaluated elementarily by acute toxicity test in mice.(2) Rats were divided randomly into five groups, model control group (distilled water,7.5ml/kg), omeprazole group (0.018g/kg), and groups with tested drug (setting three groups which were administrated the Chlorophyll-carotene paste from pine needles at a dose of0.75,1.5,2.25g/kg.), respectively. All rats were separately administered once every day for5or7days each group. Models of gastric ulcer were induced by pyloric ligation, ethanol and acetic acid in rats, and then the effect of Chlorophyll-carotene paste from pine needles on gastric lesions by evaluating the ulcer area, percentage inhibition of ulcer, gastric juice volume, PH of gastric juice, pepsin activity and the pathology change of gastric mucosa.(3) The method of doubling dilution in tubes was used in susceptibility test, the minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined.(4) Helicobacter pylori (Hp) repeat infected ethanol burning type gastric ulcer rats were used to evaluate the effect of Chlorophyll-carotene paste from pine needles on gastric lesions by examining the ulcer area, percentage inhibition of ulcer, levels of MDA (malonaldehyde), GAS (gastrin), PGE2(ProstaglandinE2) and SOD (superoxide dismutase) in blood serum and gastric tissue, and pathology change of gastric mucosa.(5) Gastric cancer cell SGC-7901were treated with different volum fractions of Chlorophyll-carotene paste-contained serum (10%,20%adnd40%), then the Gastrin promoter activity and gastrin mRNA level were detected by Real-time PCR and a Dual-Luciferase reported assay.Result:(1) Maximum tolerable dose (MTD) of the Chlorophyll-carotene paste from pine needles for mice by intragastric administration was20.0g/kg, which suggests median lethal dose (LD50) of the fraction is more than20.0g/kg.(2) The Chlorophyll-carotene paste from pine needles could relieve gastric mucosa lesions significantly in dose dependence of three ulcer models, and the effect of2.25g/kg dosage group was similar to or better than omeprazole group. In ulcer model by pyloric ligation, all of three dosages of Chlorophyll-carotene paste from pine needles could reduce gastric secretion, pepsin activity and inprove PH of gastric juice in dosage dependence, meanwhile, the effect of2.25g/kg dosage group was similar to omeprazole group. (3) Chlorophyll-carotene paste from pine needles had the inhibitory effects on Hp, with MIC of15.625mg/ml, and MBC of62.5mg/ml.(4) In Hp repeat infected ethanol burning type gastric ulcer rats, we found that all of three dosages of Chlorophyll-carotene paste from pine needles could reduce the ulcer area in dosage dependence, and the percentage inhibition of ulcer were31.47%,45.54%and53.96%, respectively. Meanwhile, the effect of2.25g/kg dosage group was similar to omeprazole group. In addition, all of three dosages could increase the levels of PGE2and SOD, and reduce MDA and GAS of blood serum and gastric tissue in dosage dependence; meanwhile, the effect of2.25g/kg dosage group was similar to omeprazole group.(5) The results indicated that Chlorophyll-carotene paste could inhibit the activity of gastrin promoter and down-regulate gastrin mRNA expression in SGC-7901cell.Conclusions:(1) LD50of the Chlorophyll-carotene paste from pine needles is more than20.0g/kg, which suggests the toxicity of Chlorophyll-carotene paste from pine needles might be minor.(2) The Chlorophyll-carotene paste from pine needles relieves in a dose-dependent way gastric lesions induced by pyloric ligation, ethanol and acetic acid in rats. It may due to the reduction of gastric secretion, pepsin activity and the content of MDA and GAS in blood serum and gastric tissue, improve PH of gastric juice and the levels of PGE2and SOD in blood serum and gastric tissue, and the antibacterial activity of Hp. The antiulcer mechanism of the fraction might be associated with attenuating aggressive factor of gastric mucosa, increasing blood flow of gastric mucosa and enhancing the effect against lipid peroxidation and of scavenging free radical, and so on. |
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