| Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used for their analgesic, antipyretic, anti-inflammatory and antiplatelet effects. Gastrointestinal symptoms are the most common adverse events associated with NSAID therapy In short or long term NSAID therapy, gastric ulcers, bleeding, and perforation are serious side effects that are observed and these ulcerative lesions are the major limitation to their use as anti-inflammatory drugs.Cuttlebone as traditional Chinese medicine, has a long history and wide application. It was used in the treatment of gastric hyperacidity, stomach ulcers and a variety of bleeding. Clinical studies have showed that cuttlebone was effective to treat peptic ulcer, but the treatment mechanism is still needed to study. Also the efficiacy of cuttlebone in treating gastric mucosal injury induced by NSAIDs. hasn’t studied.In the present study, we firstly established the model of indomethacin-induced gastric mucosal lesion in rats. Then we observed the efficiacy of cuttlebone with different doses in this model and measured the amount of EGF, PGE2, MDA and SOD.Material and methods1. Laboratory animals:60healthy, sexual-mature, and male SD rats were adpopted to our research. The body mass was150±15g.2. Division:The rats were randomly divided into6groups,10rats in each group. Cuttlebone was administered intragstrically at3000,1500and750mg/kg dosages in distilled water to groups of rats fasted for24h. At the same time,4mg/kg omeprazole in phosphate buffer was given to a fourth group of rats fasted for24h. After30min,40mg/kg indomethacin was given to each rat in all groups (3000, 1500and750mg/kg cuttlebone groups,4mg/kg omeprazole group and indomethacin control group) by oral gavage in distilled water. An identical volume of distilled water was given to the healthy group.3. Methods3.1Lesion index:We observed the gastric specimens and measured the lesion index with vernier caliper according to the Guth standard.3.2Pathological inflammation score:The gastric tissue was fixed in10%formalin, paraffin sections, HE staining, and scored according to Rauws classification standard.3.3Blood EGF and PGE2:measured by enzyme linked immunosorbent assay according to the kit.3.4Tissue PGE2, MDA and SOD:The concentration of PGE2, MDA and inhibition rate were measured by ELISA, TBA and WST-1, then divided by the prtotein concentration in tissue measured by BCA method.Result1. General observation:The control group:gastric mucosa was smooth, ruddy color, and the texture of mucosal folds were clear; the model group:gastric cavity were hemorrhagic effusion, mucosal surface with a large number of blood scab, the mucosa was edema and diffuse linear ulcer and punctate erosions after ice physiological saline flushing; the omeprazole group and cuttlebone high, middle dose group:only a small amount of punctate erosions, mild mucosal hyperemia; cuttlebone low dose group:showed gastric mucosal hyperemia and punctum ulcer lesions.2. Lesion index (LI):The LI in the model group (31.50±10.60) was higher than the omeprazole and cuttlebone high, middle dose group (17.90±3.50,18.30±6.95and20±7.13)(p<0.01). The LI in the low dose group of cuttlebone was (24.50±6.79), was decreased significantly compared with model group (p<0.05).3. Inflammation score:The inflammation score of the model group (4.70±1.16) was significantly higher than the control group(1.70±0.82)(p<0.01). The inflammation score of omeprazole group (3.20±1.32) and high dose group of cuttlebone (3.30± 1.42) were lower than that of the model group (4.70±1.16)(p<0.01). That of middle dose of cuttlebone (3.40±1.08) was lower than the model group(p<0.05). The inflammation score of the low dose of cuttlebone was also lower than the model group, but the difference was not statistically significant (p>0.05).4. The serum level of EGF:The serum EGF level of the model group (76.21±11.58pg/ml) was lower than the control group (90.47±18.97pg/ml)(p<0.05). Compared with the model group (76.21±11.58pg/ml), the omeprazole group (92.31±13.79pg/ml) was increased (p<0.05), the high, middle and low dose of cuttlebone group (111.42±13.33pg/ml,97.01±14.49pg/ml,117.83±15.36pg/ml) wereincreasec significantly (p<0.01).5. The serum level of PGE2:The serum PGE2level of the model group (100.63±32.78pg/ml) was lower than the control group (111±16.1lpg/ml), but the difference was not statistically significant (p>0.05). The omeprazole group and cuttlebone each dose group (139.60±30.67pg/ml,144.61±20.86pg/ml,136.86±21.70g/ml,163.30±34.58pg/ml) were significantly higher than the model group(100.63±32.78pg/ml)(p<0.01).6. The tissue level of PGE2:The tissue PGE2level of the model group (12.11±2.10pg/mgprol) was significantly lower than that of the control group (16±3.78pg/mgprol)(p<0.01). The omeprazole group and cuttlebone high, middle dose group (15.74±2.50pg/mgprol,15.94±3.56pg/mgprol,16.77±2.79pg/mgprol) were significantly higer than the model group (12.11±2.10pg/mgprol)(p<0.01). The cuttlebone low dose group (15.45±2.94pg/mgprol) was higher than the model group (p<0.05).7. The tissue content of MDA:The MDA content of the model group rats (0.32±0.06nmol/mgprol) was significantly higher than that of the control group (0.22±0.04nmol/mgprol)(p<0.01). The omeprazole group and cuttlebone each dose group (0.21±0.03nmol/mgprol,0.26±0.03nmol/mgprol,0.25±0.05nmol/mgprol and0.25±0.05nmol/mgprol) were significantly lower than the model group (0.32±0.06nmol/mgprol)(p<0.01)).8. The activity of SOD in tissue:The activity of SOD in the model group (1.50±0.20U/mgprol) was obviously lower than that of the normal control group (2.05±0.29U/mgprol)(p<0.01). The omeprazole group and cuttlebone each dose group were all increased compared with the model group, but only the difference between the cuttlebone high dose group (2.51±0.38U/mgprol) and the model group was statistically significant (p<0.01).ConclusionIn conclusion, this study reveals that cuttlebone showes a a curative effect against indomethacin-induced gastric lesion at all doses and the antiulcerative effect shows a positive correlation with dosage. Consequently, we can say there is a correlation between the antiulcer mechanism and increased secretion of EGF and PGE2, prevention of lipid peroxidation, activation of radical scavenging enzymes. |
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