Puerarin, an isoflavone compound extracted from Gegen (Radix Puerariae Lobatae), modulates sclera remodeling caused by extremely low frequency electromagnetic fields

OBJECTIVE: To evaluate the protective effect of puerarin [an isoflavone compound extracted from Gegen(Radix Puerariae Lobatae)] in scleral remodeling induced by extremely low frequency electromagnetic fields(ELF-EMFs).METHODS: Human fetal scleral fibroblasts(HFSFs)were divided into 5 groups:(a) untreated controls;(b) cells treated with ELF-EMFs;(c) cells treated with ELF-EMFs and puerarin 0.1 μM;(d) cells treated with ELF-EMFs and puerarin 1 μM;(e) cells treated with ELF-EMFs and puerarin 10 μM. Cell proliferation activity was measured by the cell-counting kit-8 assay. Matrix metalloproteinase-2(MMP-2) activity was measured by gelatin enzymography.MMP-2 and collagenⅠ(COL1A1) m RNA, protein expression were measured by Real-Time polymerase chain reaction, Western blot analysis, respectively.RESULTS: Puerarin reduced the inhibition in cell proliferation, MMP-2 activity, m RNA, protein expression of HFSFs exposed to ELF-EMFs and enhanced the COL1A1 m RNA and protein expression.CONCLUSION: Puerarin was found to participate in the matrix remodeling process. It might be a potential agent for the treatment of extracellular matrix degradation of sclera associated with ocular conditions.

葛根产地、品种、采收、加工及其与药材品质关系研究进展

通过检索中国知网、万方数据知识服务平台、重庆维普等数据库,总结产地、品种、采收、加工对葛根药材品质影响的研究进展,为制定葛根药材药材种植、采收、产地加工等规范提供理论依据。

Chemical Constituents from Pueraria lobata

Pueraria lobata ( Wild. ) Ohwi ( Ye-Ge in Chinese ) is a perennial herb ofthe genus Pueraria, which belongs to the Leguminosae family. It is commonly employed to relievefever and dysentery, promote the production of body fluid, reduce stiffness and pain of the nape,and for the treatment of cardiovascular diseases, e. g. hypertension, myocardial infarction, andarrhythmia. Previous phytochemical studies on P. lobata reported a number of bioactive isoflavones,e. g. daidzein, daidzin, and puerarin. Further investigation of its root has led to isolation offourteen compounds and their structures were identified as daidzein, ononin, daidzin, 3'' -methoxypuerarin, puerarin, pueroside B, daidzein-8-C-apiosyl- (1-6)-glucoside, 3''-hydroxy-puerarin,puerarinxyloside, daidzein-7, 4'' -O-glucoside, puerarin-4''-O-glucoside, mirificin-4''-O-glucoside,sissotorin, and pueroside C. Compounds 11 and 13 were isolated from the root of P. lobata for thefirst time.

超声波辅助提取葛根黄酮的工艺研究

采用超声波提取法,研究了乙醇浓度、固液比、超声波提取时间、超声波功率4个因素对葛根(Puerariae lobata(Willd.)Ohwi)黄酮提取率的影响,并采用L8(27)正交试验确定超声波提取葛根黄酮的最优工艺。结果表明:超声波作用时间与固液比对葛根黄酮得率有显著影响(P<0.05),并且其交互影响显著(P<0.05)。优化的葛根黄酮提取条件为:乙醇浓度70%、料液比1∶25、超声功率240W、超声时间40min。

正交试验优化葛根中葛根素的微波提取工艺

以体积分数50%的乙醇为提取剂,采用微波辅助法提取葛(Pueraria montana var.lobata)根中的葛根素,设计正交试验优化提取工艺。结果表明,优化的提取条件为料液比1∶40(m∶V,g/mL)、微波功率500W、微波处理时间5 min。此工艺条件下葛根中葛根素的提取率为3.51%。与加热回流法相比,采用微波辅助法葛根素的提取率提高了19.79%,提取时间缩短了90%。

高黄酮含量野葛淀粉提取工艺优化

采用单因子和正交试验方法对富含黄酮的野葛(Pueraria lobata)淀粉提取工艺进行了研究,对水、甲醇、乙醇3种溶剂提取效果进行了比较,以淀粉中的总黄酮含量为参考指标,分别对4个提取参数(提取温度、乙醇体积分数、提取时间、固液比)进行单因子试验,然后采用正交试验对提取条件进行优化。结果表明,野葛淀粉高黄酮含量提取的工艺条件为提取温度35℃、乙醇体积分数50%、提取时间3.0 h、固液比1∶8(kg∶L)。

超声波辅助半仿生法提取地桃花多糖

研究超声波法辅助半仿生法提取地桃花(Urena lobata L.)中多糖的最佳工艺。采用半仿生提取,在超声波协同作用的基础上,以正交试验法,考察超声功率、超声时间、超声温度、料液比4个因素对多糖提取率的影响。结果表明,超声波辅助半仿生法提取地桃花多糖的优化工艺条件为超声功率60 W、提取时间50 min、提取温度75℃、料液比1∶30(m/V,g∶mL)。在此条件下多糖提取率为12.86%,加样回收率为95.3%,超声波辅助半仿生提取地桃花多糖是一种可行的方法。

Ultrasonically Assisted Extraction of Isoflavones from Stem of Pueraria lobata （Willd.） Ohwi and Its Mathematical Model

Ultrasonically assisted extraction of isoflavones from the stem of Pueraria lobata （Willd.） Ohwi has been carried out with an ultrasonic extracting apparatus （20kHz, electrical power input to the transducer in 0-650W）. The influence of the electrical power input and extraction time on the＇extraction yield is investigated in water, n-butanol, and 95% （by volume） and 50% （by volume） ethanol aqueous solution. The experimental results indicate that the yields of total isoflavones are higher in ultrasonically assisted extraction than those obtained from con-ventional extraction.Moreover,a mathematical model is proposed,by introducing the electrical power input to index the ultrsound intensity,to describe the behavior of ultrasonically assisted extraction.It is found that the model calcuations are in good agreement with the experimental data.

Kinetics of Leaching Flavonoids from Pueraria Lobata with Ethanol

The kinetics of leaching flavonoids from Pueraria Lobata with ethanol was investigated. The effects of leaching temperature, mechanical agitation rate, concentration of ethanol and feed particle size on leaching kinetics were examined. It is found that the smaller the feed particle size or the higher the leaching temperature, the higher the leaching rate. The leaching process can be described by the shrinking-core model. The apparent activation energy is 10.8kJ·mol^-1, suggesting that the leaching process is controlled by the inner diffusion. An empirical equation relating the flavonoids leaching rate constant to the feed particle size and leaching temperature was expected.

Two new C-glucofuranosyl isoflavones in puerarin injection

Two new C-glucofuranosyl isoflavones of impurities in puerarin injection were isolated. Their structures were detemained to be 8-C-α-glucofuranosyl-7, 4P-dihydroxyisoflavone and 8-C-β-glucofuranosyl-7, 4＇-dihydroxy- isoflavone by chemical and spectral analysis.

Anti-diabetic potential of Urena lobata leaf extract through inhibition of dipeptidyl peptidase IV activity

Objective: To evaluate the anti-diabetic potential of leaf extract from Urena lobata(U. lobata) through dipeptidyl peptidase IV(DPP-IV) inhibitory activity.Methods: U. lobata leaf was extracted in hot water and ethanol. The activity of DPPIV inhibitor was tested by in vitro study using gly-pro-p-nitroanilide as substrat of DPPIV and vildagliptin, as standard reference. A product of the reactions between gly-pro-pnitroanilide and DPP-IV, was observed by microplate readers with λ = 405 nm. All data were expressed as mean ± SD and the IC50 value was determined by non linear regression curve fit. Active substances in leaf extract of U. lobata was analyzed by liquid chromatography-mass spectrometry. DPP-IV inhibitory activity of active compounds was evaluated in silico using docking server. Results: The ethanolic extract of U. lobata showed stronger DPP-IV inhibitor activity than water extract with the IC50 values of 1 654.64 and 6 489.88 μg/mL, respectively. Vildagliptin, based on standard reference for DPP-IV inhibitor activity, has IC50 value of 57.44 μg/mL. Based on in silico analysis, mangiferin, stigmasterol and β-sitosterol in U. lobata extract have a strong inhibitory activity on DPP-IV. Conclusions: The results showed that DPP-IV inhibitory activity of U. lobata is related to its active compounds such as mangiferin, stigmasterol and β-sitosterol.

Kinetics of Leaching Flavonoids from Pueraria Lobata with Ethanol

The kinetics of leaching flavonoids from Pueraria Lobata with ethanol was investigated. The effects of leaching temperature, mechanical agitation rate,concentration of ethanol and feed particle size on leaching kinetics were examined.It is found that the smaller the feed particle size or the higher the leaching temperature,the higher the leaching rate. The leaching process can be described by the shrinking-core model. The apparent activation en- ergy is 10.8kJ·mol-1, suggesting that the leaching process is controlled by the inner diffusion. An empirical equa- tion relating the flavonoids leaching rate constant to the feed particle size and leaching temperature was expected.

Effects of Rare Earth Elements on Promoting Isoflavonoid Production and Release of Pueraria Lobata Hairy Roots in Vitro

The effects of various rare earth elements on growth and isoflavonoid production in hairy root cultures of Pueraria lobata (Willd.) Ohwi (strain TR2) cultured in 500 ml flasks were studied. After 32 days of culture, the biomass of hairy roots increase 15 times and reach 3.2 g dry weight. Hairy root growth was inhibited by Y_2O_3, NaSeO_3 and Sm^(3+) because of brown formation. But significant promoting effect on root growth due to callus formation was observed in La^(3+) treatments. It is the most noteworthy that the production of total isoflavonoids and puerarin was enhanced greatly by La^(3+) treatment. A major portion of increased total isoflavonoids and puerarin was released into medium in La^(3+) treatment while the hairy root viabilities were preserved. Some specific secondary metabolite release processes could be induced by La^(3+) and their possible mechanism is discussed.

Removal of competitive native species combined with tree planting can accelerate the initial afforestation process: an experiment in an old field in Japan invaded by dwarf bamboo and kudzu

Restoring natural forests after field abandon- ment is a land management objective that fosters the re- covery of forest biodiversity. We performed seeding and transplanting of native tree species 40 years after the abandonment of an arable field that became dominated by a dwarf bamboo （Pleioblastus chino （Franch. et Sav.） Makino） and by kudzu （Pueraria lobata （Willd.） Ohwi）. By permutation tests, the removal of competing vegetation （gap creation） significantly increased the survival of three seeded species of Fagaceae and of eight transplanted spe- cies. In contrast, intact vegetation prevented most indi- viduals of all species from surviving for 1 year after planting. The lack of natural recruitment of Fagaceae in the nonseeded subplots indicated that seed limitation was a cause of the slow afforestation. Although litter accumula- tion in gaps at the time of seeding slightly increased sur- vival for late-germinating Quercus myrsinifolia Blume andCastanopsis sieboldii （Makino） Hatus. ex T. Yamaz. et Mashiba, the effect was not consistent among plots and was not statistically significant. Our results suggest that for successful afforestation using native trees in abandoned fields, it will be necessary to remove competitive native species to avoid severe limitations on microsite availability and that simultaneous tree establishment by seeding or transplanting should be implemented to accelerate the establishment of native tree species.

Study on the Production Technology of Pueraria lobata Jelly

An orthogonal experiment was carried out to investigate the optimal production formula of Pueraria lobata jelly combining P. lobata,red beans and sugar as the main raw materials. The isoflavone contents in P. lobata and the P. lobata jelly were determined by colorimetry and sensory evaluation. The results showed that the optimal formula for the product was: P. lobata paste 50%,red bean paste 60%,and white granular sugar and agar 20% and 3% of the total weight of P. lobata paste and bean paste,respectively. The development of this product is of great significance to the utilization of P. lobata.

Estimating plant crown transpiration and water use efficiency by vegetative reflectance indices associated with chlorophyll fluorescence

This research developed estimates of plant crown transpiration and water-use-efficiency using reflectance and derivative indices extracted from remotely sensed chlorophyll fluorescence measurements under natural conditions. Diurnal changes of leaf-level gas exchange (carbon assimilation rate (A), stomatal conductance (gs), transpiration rate (E)), chlorophyll fluorescence and canopy-scale remote sensing were measured on top crown of valley oak (Quercus lobata) in the foothills of central California, USA. The results indicated Q. lobata experienced saturating irradiance (PAR), which induced photoinhibition indicated by a decrease in the quantum efficiency of photosystem II (r2 = 0.648 with Fv ′/Fm′ and r2 = 0.73 with FPSII) and open reaction centers (qP;r2 = 0.699). The excess absorbed quantum energy was dissipated as heat through the Xanthophyll cycle and other processes (photorespiration and the water-water cycle) rather than energy emission as steady state chlorophyll fluorescence (Fs). An increase in leaf temperature caused by the activity of Xanthophyll cycle was correlated to a decrease in Fs (r2 = 0.381) and an increase in evaporative cooling through E (r2 = 0.800) and water use efficiency (WUE;r2 = 0.872).

Comparative molecular docking studies of lupeol and lupenone isolated from Pueraria lobata that inhibits BACE1: Probable remedies for Alzheimer's disease

Objective: To discover lead lupane triterpenoid's potential isolated from Pueraria lobata roots against b-site amyloid precursor protein cleaving enzyme 1(BACE1), which serve as a rate limiting step in amyloid beta(Aβ) production altering the course of Alzheimer's disease. In addition, enzyme kinetics study and molecular docking were conducted to establish the inhibition type and structure activity relationship.Methods: A systematic study of 70% ethanolic P. lobata root extract was employed to identify its BACE1 inhibitory potential. Further, BACE1 inhibitory potential of two lupane terpenoids, yielded from ethanolic extract, was assessed. In order to determine their inhibition mode, Lineweaver–Burk plots and Michaelis–Menten model for BACE1 was performed. Auto Dock 4.2 program in addition determined the molecular interaction of BACE1 with isolated terpenoids.Results: Considering the inhibitory potential of 70% ethanolic extract of P. lobata against BACE1(IC_(50)= 80.35 mg/mL), lupeol and lupenone were subsequently isolated and exhibited notable or moderate BACE1 inhibitory activity with IC_(50) values of 5.12 and 62.98 mmol/L, respectively, as compared to the positive control quercetin(IC_(50)= 21.28 mmol/L). The enzyme kinetics study enabled us to identify both compounds as competitive inhibitors, where lupeol displayed a very potent inhibition against BACE1 with low inhibition constant(Ki) value of 1.43 mmol/L, signifying greater binding affinity.In order to understand the binding mechanism and structure–activity relationship of two triterpene-based BACE1 inhibitors, we employed computer aided docking studies which evidently revealed that hydroxyl group of lupeol formed two hydrogen bonds with the ASP32(catalytic aspartic residue) and SER35 residues of BACE1 with the binding energy of(-8.2 kcal/mol), while the ketone group of lupenone did not form any hydrogen bonds with BACE1 giving evidence for less binding affinity. These results in turn have predicted the dependence of the inhibitory activity in the presence of hydroxyl group which has provided a new basis for BACE1 blockade.Conclusions: Our results have successfully explored the molecular mechanism of lupane triterpenoids via BACE1 inhibition, suggesting that lupeol in particular could be utilized as a useful therapeutic and preventive agent to mitigate Alzheimer's disease.

Kudzu Forage Quality Evaluation as an Animal Feed Source

The nutritional value of kudzu (Pueraria lobata) was evaluated as a potential animal feed. Randomly selected samples from natural habitats with Jacksonville area (Alabama, USA) were utilized in the study. Kudzu leaves showed significantly higher dry matter and crude protein content than the stems;whereas, ADF fiber was significantly lower in the leaves. However, NDF and total digestible nutrients (TDN) were similar in the leaves and the stems. The leaf part of kudzu was especially rich in CP (24.46%) and met the requirements and recommendations for most ruminants. The TDN value for kudzu stem is 55.99%, which falls below but within the recommended range. Macronutrients concentration between the stem and leaf of kudzu, nitrogen, calcium, and magnesium showed a significantly higher in the leaf than the stem. However, phosphorous was similar in concentration in the stem and the leaf, and potassium was higher in the stem. In regard to the micronutrients, the only significant difference between the leaf and the stem was iron. However, iron, manganese, zinc, and copper were all above the recommended concentrations by the National Research Council. In general, kudzu can be considered a good source for animal feed, especially the leaf, which shows a higher value in most of the determined criteria in comparison to what was recommended by the National Research Council. This should provide a new outlook to kudzu, which has been considered a noxious weed and help in the utilization of kudzu in a positive way in the southern region of the US.

十堰市葛根资源利用现状及发展建议

介绍了葛(Pueraria lobata)根的营养成分及功效,分析了十堰市葛根资源利用现状及发展前景,并针对葛根产业发展存在的主要问题,提出了十堰市葛根产业的发展建议。

Diurnal changes in net photosynthetic rate of Pueraria lobataand its impact factors

In order to identify the relationship between diurnal changes in the net photosynthetic rate（Pn） of Pueraria lobata and environmental factors,diurnal changes in the Pn of leaves of two P.lobata cultivars were measured using a CIRAS-1 portable photosynthesis measurement system（PP-Systems,UK）.The results show that diurnal changes in Pn of both cultivars could be interpreted as double-peak curves,indicating the occurrence of an obvious midday depression.Further analyses indicate that the correlation between Pn and stomatal conductance was positive and extremely significant（p 〈 0.01）.The correlations of Pn with intercellular CO2 concentration and transpiration rate were positive and significant（p 〈 0.05）,while the correlations of Pn with air and leaf temperatures were negative and significant（p 〈 0.05）.The results indicate that among the factors affecting photosynthetic properties,some can be grouped as stomatal limitations while others are non-stomatal limitations.