| Objective: Bauhinia championii(Benth.)Benth.,which belongs to Leguminosae,Caesalpinioideae,Benth and its medicinal part is the dry stem.It is a kind of traditional Chinese medicine widely distributed in southeast Asian countries such as Vietnam,Sri Lanka,Indonesia and China’s Guangdong,Guangxi,Fujian,Taiwan,Guizhou,Hunan and so on.It is neutral in nature,sweet in taste with slightly bitter and the channel tropism manifests in its therapeutic action on liver and large intestine.Therefore,Bauhinia championii has been proved curative effect in clinically treating rheumatism,apoplectic hemiplegia,stomachache,infantile malnutrition.In TCM theory,gastric ulcer(GU)is assigned to the field of epigastralgia,TCM and its preparation used to treat epigastralgia are effective in treatment of GU with unique advantages and prominent effect.For this reason,it’s of momentous significance and high feasibility to choose active constituents or effective fractions which are of curative effect in treating GU.On one hand,this essay expounds that macroporous resin can be applied to enrich polymethoxyflavones from Bauhinia championii(PMF)and the more optimal technology of purifying PMF.On the other hand,in order to provide certain experimental foundation for clinical application and drug development of PMF,researcher make a preliminary investigation on therapeutic action of GU by total polymethoxyflavones from Bauhinia championii(PMFS),which develops effective drugs to cure GU,and satisfies the needs of market.Methods:(1)Purification technology research of Bauhinia championii(Benth.)Benth.HPLC was adopted to determine four polymethoxyflavones and purification technology of macroporous resin was optimized using single factor.(2)Study on the protective effects of PMFS against experimental gastric ulcer.Phase one of this study,three kinds of gastric ulcer model were used to observe the efftect of the polymethoxyflavones,which include type of ethanol,indomethacin,water immersionrestraint strees in mice.Animals which were randomly divided into model groups,PMFS with low,medium and high dose group(3.9,7.8,15.6 mg·kg-1),omeprazole enteric-coated capsules group(5 mg·kg-1),ranitidine hydrochloride capsules group(50 mg·kg-1).Each ulcer area of mice were measured and calculated ulcer inhibition rate.Phase two of this study,to investigate the protective funetion of PMFS against gastric ulcer in ethanol rat.Animals which were randomly divided into model groups,PMFS with low,medium and high dose group(2.1,4.2,8.4 mg·kg-1),omeprazole enteric-coated capsules group(5mg·kg-1),ranitidine hydrochloride capsules group(50 mg·kg-1).Acute gastric ulcer model in rats were induced by ethanol(5 m L·kg-1),and then the effect of PMFS on gastric lesions by evaluating the ulcer area percentage inhibition of ulcer.At the same time,serum inflammatory mediators TNF-α,IL-6 and IL-10,gatric oxidative status SOD,GSH,CAT and MDA were measured to explain the possible protective mechanisms of total polymethoxyflavone in ethanol-induced gastric ulcer.(3)To measure median lethal dosage of PMFS with method of Bliss.Results:(1)It showed a good purifying profile on NKA resin with the conditions as follows: the concentration of sample extract was 0.66 g·m L-1,the amount of saturated adsorption at 5 body volumn(BV),the flow rate was 2 BV·h-1,then eluting successively with 5 BV water and 8 BV 35% ethanol solution to remove impurities,elution with 75% ethanol at length.75% ethanol eluate was collected and dried.Eventually,the total mass fraction of four polymethoxyflavones in the solid sample was 36%.(2)The present study showed total polymethoxyflavones exerted gastric-protective effect anti-experimental gastric ulcer on the three ulcer models,which could significantly reduce the ulcer area of the mice and had a high inhibition rate.Total polymethoxyflavones of any dosage prior to ethanol ingestion effectively protected the stomach from ulceration and decrease the percentage inhibition of ulcer(P<0.05).(3)The total polymethoxyflavones in different doses could significantly reduce the ulcer area of the ethanol-induced rats(P<0.01)and had a high inhibition rate(49.35%,52.53%,66.63%),and improve pathological morphology.Pre-treatment with total polymethoxyflavones could elevate the level of IL-10(P<0.01),while markedly mitigate the concentration of IL-6(P<0.05)and TNF-α(P<0.01 or P<0.05).Compared with the model group,total polymethoxyflavones groups could increase the SOD activity of rat’s gastric tissue(P<0.01),while decreased the MDA concentration of rat’s gastric tissue(P<0.05 or P<0.01).The high dose of total polymethoxyflavones could increase the CAT activity of rat’s gastric tissue as well as rat’s gastric GSH concentration(P<0.01).(4)The LD50 of total polymethoxyflavones was 1256.319 mg·kg-1.Conclusion:(1)NKA-type macroporous resin can be used for the enrichment of polymethoxyflavone in Bauhinia championii(Benth.)Benth,and the purification process is stable and feasible.(2)The total polymethoxyflavones from Bauhinia championii(Benth.)Benth has obvious protective effect on experimental gastric ulcer models in mice.(3)The total polymethoxyflavones in different doses can significantly reduce the ulcer area of the ethanol-induced rats and has a high inhibition rate,and improve pathological morphology.It can decrease the levels of TNF-α and IL-6,increase the level of IL-10,increase the activities of CAT,SOD and GSH in gastric tissue,and decrease the content of MDA in gastric tissue of rats.The present study showe PMFS exerted gastric-protective effect anti-experimental gastric ulcer caused by ethanol and the mechanism is probably related with suppressing regulation of attack factors and enhancing protective factors.(4)The total polymethoxyflavones have a certain extent of acute toxicity in mice. |
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