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Objective: ① off Cangzhu different extract fractions of ethanol-induced gastric mucosal injury experiment the screen mainly active site, and to explore the mechanism of the protective effect of off herb active site of ethanol-induced gastric mucosal injury prevention; ② by GC-MS technical analysis determination of the relevant herb Supercritical extract chemical composition and HPLC method off herb supercritical extract atractylenoide Ⅰ, atractylenoide Ⅲ content determined off the herb in the active site of the gastric mucosal protective effect of prevention may ingredients. Methods: ① Supercritical-C02 Extraction off herb supercritical extraction of material (GCC), the dregs after the extraction extraction of ethanol as entrainer continue under the same conditions, the addition of ethanol entrainer supercritical extract (GCCY ). The supercritical extraction dregs water extraction and extracted with n-butanol extract the water layer (GCS) and positive butanol layer (GCZ) of two parts. Establishment of ethanol-induced gastric ulcer model in mice, oral administration of the four extract, administered for 5 consecutive days, measuring the gastric ulcer area and gastric tissue content of NO, PGE2, and explore the four extract the stomach mucosal injury, and to identify the extract fractions from the gastric mucosal protective effect GCCY. (2) ethanol-induced rat gastric ulcer model, respectively, were given 0.35 g / Kg / d and 0.70 g / Kg / d of GCCY administered for 5 consecutive days, measuring the gastric ulcer area and stomach tissue MDA, MPO, NO content and SOD, NOS activity, and determination of COX-2 protein expression, to further explore the extraction of parts from the main mechanism of gastric mucosal protection. ③ the two supercritical extracts using GC-MS determination of customs herb, identified two main component of the extract compare. The GC conditions inlet temperature 200 ° C, the starting temperature of 40 ° C, retained for 1 min, 5 ℃ / min, the temperature was raised to 140 ℃, 10 ℃ / min, the temperature was raised to 250 ° C and retained for 5 min, carrier gas He ; split ratio of 50:1, the constant current of 1.00 mL / min, ionization mode EI, ionization voltage 70 eV, connecting rod temperature of 280 ℃, the ion source temperature of 280 ° C, mass spectral libraries NIST8.0LIB. (4) using high performance liquid chromatography (HPLC) method the the determination off herb extract atractylenoide Ⅰ atractylenoide Ⅲ content, chromatographic conditions as the mobile phase was methanol - acetonitrile (60:40), the detection wavelength was 210 nm, a flow rate of 1.0 mL / min, column temperature at room temperature. Results: ① Compared with model group, three off the herb extract GCCY, GCC and GCZ could significantly reduce the mouse gastric ulcer index (P lt; 0.01), the gastric membrane GCCY group NO content was significantly increased (P lt; 0.05) GCCY group and the GCC group mucosal PGE2 content increased significantly. ② GCCY0.35 g / Kg / d and 0.70 g / Kg / d dose group and the model group compared to both a reduction in the extent of gastric mucosal damage area. Compared with the model group, the dose group GCCY two MDA (P lt; 0.01) and MPO were significantly lower (P lt; 0.05), while SOD was significantly higher (P lt; 0.05); raised in rats high doses of ethanol gastric tissue COX-2 expression, COX-2 expression and GCCY 0.35 g / Kg / d dose group was significantly lower. ③ by GC-MS technique of GCC and GCCY, detect, divided distinguish peaks 71 and 49, 53 and 25 components were identified, peak area normalization method for the determination of their relative percentage of the total contents accounted the GCC, GCCY total of 98.56% and 94.33%, respectively. Compared with the GCC, GCCY a unique ingredient, ethyl acetate. The ④ HPLC measured atractylenoide Ⅰ atractylenoide Ⅲ the content than GCCY in the GCC. Conclusion: ① Off supercritical extract of the herb GCC, GCCY ethanol-induced gastric mucosal injury have preventive and protective effect, including a strong protective effect in GCCY. GCCY possible through the lower MPO and free radical damage biofilm;, lowered the COX-2 gene expression by maintaining the level of the gastric mucosa NO, play an important role in the prevention of gastric mucosal injury. The ② GC-MS, respectively 53 and 25 ingredients identified in the GC, GCCY. Compared with the GCC, GCCY impurities; unique ingredients as ethyl acetate, ethanol entrainer into four components relative percentage significantly higher than the GCC. The ③ Atractylenoide Ⅰ and atractylenoide Ⅲ content is higher than in the GCC GCCY, off the herb may also exist outside the atractylenoide Ⅲ gastric mucosal protective ingredients, GCCY than GCC higher relative percentage composition may also gastric mucosal protective ingredients.
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