| Alpiniae officinarum Rhizoma(AR)is the dry rhizome of Zingiberaceae,which has the effects of warming stomach and dispelling cold,and relieving digestion and pain.In this paper,the research progress of the main chemical components,pharmacological effects and gout of AR was reviewed,which laid a literature foundation for the anti-gout experiment of AR active parts.First,the AR was separately subjected to water extraction and alcohol extraction,and the alcohol extract was sequentially subjected to system solvent extraction with petroleum ether,dichloromethane,ethyl acetate and n-butanol.The hyperuricemia model of mice was established by intragastric administration of xanthine and intraperitoneal injection of potassium oxonate.The serum uric acid value was detected and the uric acid-lowering active site of AR was initially screened.The results showed that the n-butanol extraction of AR could significantly reduce the serum uric acid level in mice,followed by alcohol extract,mother liquor,water extract and other parts.These results indicated that there were strong alcohol soluble and water soluble active ingredients in AR to reduce the level of uric acid.In particular,the aqueous extract mainly contains polysaccharides and volatile oils,while the n-butanol fraction contains a large amount of flavonoid glycosides.Therefore,it is proposed to carry out extraction and purification and activity screening studies on these parts.Secondly,the volatile oil of AR was extracted by steam distillation.The yield of volatile oil was used as the index.Four factors including the ratio of material to liquid,pulverization degree,soaking time and extraction time were selected to investigate and screen out the best extraction process.The results showed that the highest yield of the 24 target powder of AR was 0.171 mL/g under the conditions of 1?12(g/mL),1hour of soaking and 5 h of extraction.The polysaccharide of AR was extracted by water and precipited by alcohol.The content was determined by the method of phenol-sulfuric acid.The extraction yield and content were used as indexs in Single factor and orthogonal experiments to select the ratio of material to liquid,extraction temperature and extraction time.The optimalconditions for the extraction of polysaccharides from AR were as follows: the ratio of material to liquid was 1?45(g/mL),the extraction temperature was 100?,and the extraction time was 2.5 h.Under these conditions,the content and yield of polysaccharide were 39.83% and 52.71%,respectively.The total flavonoids of AR were extracted by heating and refluxing.The method of ultraviolet spectrophotometry to determine the total flavonoids content of AR was established.The yield and content of total flavonoids were used as indexs in Single factor and orthogonal experiments to optimize the best extraction conditions.And then,the purity of Galangin was determined by high performance liquid chromatography.Finally,the optimal extraction conditions for the total flavonoids of AR were: material-to-liquid ratio was 1?6(g/mL),extraction temperature was 60?,extraction time was 1.5 h,ethanol concentration was 85%.Under this condition,the yield of total flavonoids of AR was 14.69%,the purity was 22.69%,and the purity of Galangin was 5.21%.Different types of macroporous resins were investigated and AB-8 macroporous resin was screened out according to the static adsorption experiment.Then,the conditions of loading volume,pH value of the sampled solution and concentration of elution solvent were investigated by dynamic experiments,and a method for efficiently separating and purifying the total flavonoids from the alcohol extraction of AR was determined.The results showed that the optimal conditions were:pH=5 of the sample solution,2.5 g/g of dynamic saturated adsorption,and 80%ethanol.While the purity was 96.93%,and the yield was 66.28% by using AB-8macroporous resin combining with polyamide resin to purify the total flavonoids of AR.Thirdly,The mouse hyperuricemia model was used to investigate the effects of aqueous extracts,alcohol extracts,total flavonoids,volatile oils and polysaccharides on uric acid,creatinine and urea nitrogen levels in mouse serum.The results showed that the ethanol extract and total flavonoids of AR could effectively reduce the levels of serum uric acid and urea nitrogen,the high-dose volatile oil could reduce the level of serum uric acid,the water extraction could reduce the levels of creatinine and urea nitrogen,and the polysaccharide could reduce the creatinine level.Fourthly,A mouse model of gouty arthritis was established by injection of sodium urate crystals(MSU)into the ankle joint.The toe swelling volume of C57 mice was investigated by different extraction sites of petroleum ether,dichloromethane,ethyl acetate and n-butanol.Effects of foot balance differences,serum IL-1? and TNF-? protein levels.The results showed that the extractions could relieve joint swelling and pain,and reduce the levels of IL-1? and TNF-? in serum,especially the n-butanol extraction.The effects of total flavonoids of AR with different purity were investigated in the model of sodium urate crystals(MSU)-induced mouse ankle joint.The swelling volume of the toes,the balance of the feet and the levels of IL-1? and TNF-? in serum were determined.The pathological changes of the joint were also observed by HE-staining.The results showed that the extractions could reduce the degree of swelling of the toes or the weight of the feet,and lower the levels of IL-1? and TNF-? in the serum.Moreover,the joint inflammation of mice in different groups(exclude the lower-dose group)were alleviated,especially that of in the high-dose AB-8 group and the alcohol-extracted group.Fifthly,the effects of different compounds of AR were investigated in MSU-induced airbag inflammatory mice.The leukocyte count,levels of IL-1? and TNF-? in the airbag lavage fluid were detemined.Moreover,macrophage and neutrophils in the airbag lavage fuild and the airbag skin were examined by immunohistochemical staining.The results exhibited that some compounds in AR could inhibit the levels of inflammatory factors TNF-? and IL-1? and reduce the number of white blood cells.They also could effectively inhibit the wall thickening of airbags,reduce vascular proliferation and fibrosis,and improve inflammation.In summary,the total flavonoids of AR could significantly ameliorate the hyperuricemia and gouty arthritis,and have a good development potential as anti-gout drugs. |
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