Effects of <i>Myrothecium verrucaria</i>on Ultrastructural Integrity of Kudzu (<i>Pueraria montana var. lobata</i>) and Phytotoxin Implications

The fungus Myrothecium verrucaria (Alb. & Schwein.) (MV), originally isolated from diseased sicklepod (Senna obtusifolia L.), has bioherbicial activity against kudzu and several other weeds when applied with low concentrations of the surfactant Silwet L-77. To more fully understand the initial events of MV infection or disease progression, and to improve knowledge related to its mechanism of action, the effects of MV and its product (roridin A) on kudzu seedlings were examined at the ultrastructural level. Ultrastructural analysis of MV effects on kudzu seedlings revealed a rapid (~1 h after treatment) detachment of the protoplast from the cell wall and plasmodesmata appeared to be broken off and retained in the wall. These symptoms occurred well in advance of the appearance of any fungal growth structures. Some fungal growth was observed after severe tissue degeneration (24 to 48 h after treatment), but this occurred primarily at the extra-cellular location with respect to the kudzu tissues. Kudzu seedlings treated with roridin A, a trichothecene produced by the fungus, exhibited some symptoms similar to those induced by the fungus applied in spore formulations with surfactant. The overall results are the first to report the ultrastructural effects of this bioherbicide on plants and suggest that penetration of a phytotoxic substance(s) in the fungal formulation was facilitated by the surfactant, and that roridin A exerts phytotoxicity toward kudzu.

Evaluation of the Estrogenic Activity of Pueraria (Kudzu) Flower Extract and Its Major Isoflavones Using ER-Binding and Uterotrophic Bioassays

Pueraria flower extract (PFE) is a hot water extract of the Kudzu flower (Pueraria thomsonii). Tea made from dried Kudzu flower is widely used inChina, and PFE is utilized as a nutritional supplement inJapan. PFE contains unique isoflavones such as 6-hydroxygenistein 6,7-di-O-glucoside (6HGDG), tectorigenin 7-O-xylosylglucoside (TGXG), and tectoridin. 6HGDG is known to be metabolized into 6-hydroxygenistein, and TGXG and tectoridin are known to be metabolized into tectorigenin in the digestive tract. Isoflavones typically mimic the effects β-estradiol has on estrogen receptors (ERs) and may influence the female genital system in the case of excessive intake. As a result, the upper limit of safe daily consumption of soy isoflavones has been enforced inJapan. In the present study, ER-binding assays were performed using the EnBio estrogen receptor/cofactor assay system to compare the estrogenic activity of 6-hydroxygenistein and tectorigenin to that of the soy isoflavone genistein. In addition, uterotrophic bioassays were performed to investigate the estrogenic effects of PFE in vivo. The ER-binding assays revealed that the ER-binding affinities of 6-hydroxygenistein and tectorigenin were approximately 0.01 - 0.04 that of genistein. Soy isoflavone products also induced an increase in uterine wet and blotted weight at doses of 500 mg/kg and 1000 mg/kg, whereas PFE did not cause adverse estrogenic effects, even at a dose of 1000 mg/kg. Based on these results, PFE does not appear to contain compounds with strong estrogenic activity or cause adverse estrogenic effects in vivo. Importantly, the results of this study confirm the safety of PFE as a food supplement.

不同施肥措施对岩质坡面覆土平台区葛(Pueraria lobata)生长影响的综合评价

为获得一种有利于石灰岩矿岩质坡面覆土平台区葛[Pueraria lobata(Willd.)Ohwi]快速生长的土壤基质改良措施,以无为杨家岭石灰岩矿东采区第三阶平台区已实施表层覆土的岩质坡面作为试验区,评价不同施肥措施对葛在重构土壤上的生长状况。试验小区设置:A(100%复合肥)、B(75%复合肥+25%有机肥)、C(50%复合肥+50%有机肥)、D(25%复合肥+75%有机肥)、E(100%有机肥)、CK(对照,不施肥)。2022年8月,对葛的生理生化指标进行分析,并用熵权法对葛的生长状况进行综合评价。结果表明:与CK相比,施肥措施均增加了葛的株长及其叶片的干重,提高了含水率、叶绿素含量、氮磷钾含量、可溶性蛋白质含量以及3种抗氧化酶(POD、CAT、SOD)活性,同时均降低了叶片的MDA含量。综合评价结果表明,25%复合肥+75%有机肥的处理最利于葛的生长,可用于该岩质坡面植被重建的生产实践。

Puerariae Radix protects against ulcerative colitis in mice by inhibiting NLRP3 inflammasome activation

Ulcerative colitis(UC)is a common inflammatory disease of the gastrointestinal tract.Traditional Chinese medicine(TCM)has long been used in Asia as a treatment for UC and Puerariae Radix(PR)is a reliable anti-diarrheal therapy.The aims of this study were to investigate the protective effect of PR using the dextran sulfate sodium salt(DSS)-induced UC model in mice and identify molecular mechanisms of PR action.The chemical constituents of PR via ultra-performance liquid chromatography/tandem mass spectrometry and identified potential PR and UC targets using a network pharmacology(NP)approach were obtained to guide mouse experiments.A total of 180 peaks were identified from PR including 48 flavonoids,46 organic acids,14 amino acids,8 phenols,8 carbohydrates,7 alkaloids,6 coumarins and 43 other constituents.NP results showed that caspase-1 was the most dysregulated of the core genes associated with UC.A PR dose of 0.136 mg/g administered to DSS treated mice reversed weight loss and decreased colon lengths found in UC mice.PR also alleviated intestinal mucosal shedding,inflammatory cell infiltration and mucin loss.PR treatment suppressed upregulation of NOD-like receptor protein 3(NLRP3),cysteinyl aspartate-specific proteases-1(caspase-1),apoptosis-associated speck-like(ASC)and gasdermin D(GSDMD)at both the protein and m RNA expression levels.The addition of a small molecule dual-specificity phosphatase inhibitor NSC 95397 inhibited the positive effects of PR.These results indicated that PR exerts a protective effect on DSS-induced colitis by inhibiting NLRP3 inflammasome activation in mice.

Pueraria isoflavones inhibit XOD and GLUT9 to decrease uric acid production and promote uric acid excretion, respectively

Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male Kunming mice were divided into the healthy group (carboxymethylcellulose sodium at a dose of 250 mg/kg), the model group (HUA mice were given carboxymethylcellulose sodium at a dose of 250 mg/kg), the low group (HUA mice were given pueraria isoflavone at a dose of 125 mg/kg), HUA mice were given pueraria isoflavones at a dose of 250 mg/kg once d frequency)and the high group (HUA mice were given pueraria isoflavones at a dose of 500 mg/kg once d frequency) dosage groups, with 8 mice in each group. The contents of uric acid (SUA), urea nitrogen (BUN) and creatinine (SCr) in serum and urine of each group were compared before and after intervention (30 d). Statistical differences of xanthine oxidase (XOD) and human glucose transporter 9(GLUT9), cy- clooxygenase- 2(COX-2), tumor necrosis factor (TNF-α) and interleukin-1 (IL-1β) contents in renal tissues of each group after intervention (30 d) were compared. Results: After intervention, kidney inflammatory factors (COX-2, TNF-α and IL-1β) in the model group were compared. Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were higher than those of healthy group(P<0.05). Renal inflammatory cytokines (COX-2, TNF-α and IL-1β) in low, medium and high dose groups;Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were lower than those of model group, and there were low > medium > high dose groups, the comparison between the two groups had statistical significance(P< 0.05). After intervention, the contents of 3 indicators in blood or urine(COX-2, TNF-α and IL-1β) all decreased compared with before intervention, and the differences in intra-group comparison were statistically significant (P<0.05). Conclusion: Pueraria isoflavones can treat HUA mice by inhibiting the expression of XOD and GLUT9, and then play a role in reducing uric acid pro- duction and promoting uric acid excretion, as well as alleviating the degree of disease inflammation.

Effects of Radix Puerariae, Radix Rehmanniae and Their Compatibility on Blood Glucose and Blood Lipids in Mice

[Objectives]To explore the effects of the compatibility of Radix Puerariae and Radix Rehmanniae on blood glucose and blood lipids in diabetic mouses.[Methods]Diabetic mouse model was established.The body weight and fasting blood glucose of mice were measured after 7 and 14 d of administration,and the biochemical indicators of blood lipids(TC,HDL-C,and LDL-C)were detected after 14 d of administration.[Results]Compared with the Radix Puerariae group and Radix Rehmanniae group,the compatibility group(1:2)had the best hypoglycemic effect(P<0.05),and TC and LDL-C in the compatibility group(2:1)significantly decreased(P<0.05),while HDL-C in the compatibility group(1:1)significantly increased(P<0.05).[Conclusions]Radix Puerariae,Radix Rehmanniae and their combination can reduce the blood glucose of diabetic mice.The compatibility group(1:2)had a significant hypoglycemic effect(P<0.05),and LDL-C in the compatibility group(2:1)significantly declined,while HDL-C in the compatibility group(1:1)rose significantly.

Optimization of Preparative Separation and Purification of Total Flavonoids from Radix Puerariae by Macroporous Resin Method

Aim To screen the optimum macroporous resin and conditions for the isolation and purification of flavonoids from Radix Puerariae. Methods The static and dynamic adsorption/desorption methods were used, and the separation and purification process was evaluated by measuring the concentration of total flavonoid in the fractions with UV spectrophotometer. Results The SP70 macroporous resin was the most effective compared with other macroporous resins. The optimum conditions were screened, which were 0.5 g· mL^- 1 corresponding to crude drug for concentration of extract, pH 5 - 6, and appended 60 times the volume of the resin bed （BV） with the adsorption speed 2 BV·h^-1, and the volume of aq. 70% （V/V） ethanol as eluant was 5 BV with desorption speed 2 BV·h^-1. By this method, the final contents of total flavonoids exceeded 80%. Conclusion The SP70 macroporous resin is the most effective one for large-scale isolation and purification of flavonoids from Radix Pueraria, which meets industrial needs.

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.

中国葛属（Pueraria DC．）的研究

本文对国产葛属植物９种２变种进行了全面整理．对其中的主要类群进行了深入的研究．通过野外考察，腊叶标本的研究和细胞染色体的计数．使对中国葛属植物有一个较为全面的了解。

一株耐酸野生葛藤(Pueraria lobata)根瘤菌的筛选与耐酸机理初步研究

从重庆市北碚区缙云山酸性黄壤（pH4．6）上生长的葛藤根瘤中分离到一株耐酸葛藤根瘤菌PR389，能在pH4．3的YMA培养基上正常生长，而一般根瘤菌最适生长pH值为6．5—7．5，说明PR389为一株耐酸葛藤根瘤菌。通过质子通量试验发现，与不耐酸的菌株相比，PR389的细胞膜能阻止过量的H^＋进入细胞，表明PR389具有某种能力使之在低酸性环境下不受伤害。在耐酸性试验中，PR389在加氯霉素的强酸性（pH3．8）YMA培养液中表现出来的耐酸性被氯霉素抑制。推测胞内特异蛋白质的诱导合成是PR389具有耐酸性的原因。

Protective Effect of Pueraria lobota Extracts on Mitochondria Damage of Liver

[Objective] This study aimed to investigate the protective effect of Pueraria lobota extracts on mitochondria damage of liver. [Method] The liver mitochondria injury was induced by Vc-Fe2＋ , and the influences of Pueraria lobota extracts on mitochondria ATPase activity, mitochondria swelling and protein carbonyl content were measured. In addition, the lipid peroxidation in liver mitochondria was induced by H2O2-Fe2＋ to analyze the influence of Pueraria lobota extracts on MDA content. Futhermore, NBT method was used to evaluate the inhibitory function of Pueraria lobota extracts on the superoxide anion. [Result] The results showed that Pueraria lobota extracts could significantly inhibit mitochondria oxidative damage,prevent mitochondria swelling and ATPase activity reduction,decrease protein carbonyl level,and effectively scavenge superoxide anion produced by mitochondria, indicating that Pueraria lobota extracts can protect rat liver mitochondria from oxidative damage. [Conclusion] This study provided theoretical basis for investigating the pharmacological functions of Pueraria lobota.

野葛(Pueraria lobata)愈伤组织诱导及组织培养

以野葛茎尖分生组织为外植体,通过不同激素与MS培养基配比试验,筛选出野葛茎尖分生组织愈伤组织诱导与植株再生的最适培养基,建立野葛植株再生体系。试验结果表明:茎尖分生组织在MS+2,4-D 0.5 mg/L培养基上启动诱导愈伤组织,再转移到MS+6-BA 0.5 mg/L+NAA 0.5 mg/L继续诱导,愈伤组织诱导率高,繁殖能力旺盛。在MS+6-BA 1 mg/L培养基上愈伤组织分化率100%,生长能力较强。在MS+NAA 0.1 mg/L培养基上,诱导生根率达到60%。

葛属（Pueraria DC．）的分支分析

本文采用２４个形态性状对葛属进行了分支分析，得到了１１个最简约分支图和一个严格一致化分支图，葛属的种间关系。根据分支分析结果对ｖａｎｄｅｒＭａｅｓｅｎ的葛属分类系统提出了修订意见。

葛根汤干预脊髓型颈椎病模型大鼠神经炎症反应和凋亡的作用

背景:炎症和细胞凋亡在脊髓型颈椎病病理过程中起着关键作用。既往研究表明葛根汤对脊髓型颈椎病有着良好的治疗效果。目的:探讨葛根汤对脊髓型颈椎病大鼠神经炎症反应和细胞凋亡的影响及作用机制。方法:60只SD大鼠随机分为正常组、假手术组、模型组、葛根汤低、中、高剂量组,采用吸水膨胀材料压迫脊髓的方法构建脊髓型颈椎病模型,术后2周给予4.86,9.72,19.44 g/kg葛根汤灌胃,其余组则灌胃生理盐水,每日1次,持续4周。在给药后第1,7,14,21,28天对大鼠进行运动功能评价(BBB评分);在给药4周后采用苏木精-伊红染色观察脊髓组织病理形态变化,免疫荧光双染检测脊髓组织小胶质细胞极化情况,荧光定量PCR检测脊髓组织中白细胞介素6和白细胞介素1β mRNA表达,Western blot法检测脊髓组织中p-NF-κB p65、NF-κB p65、NLRP3、ASC、Cleaved Caspase-1、Bax、Bcl-2、Cleaved Caspase-3、NOX4、p-Drp1、Drp1和Mfn2蛋白表达,TUNEL法检脊髓组织细胞凋亡情况,DHE染色检测脊髓组织中活性氧水平。结果与结论:(1)与正常组、假手术组比较,模型组大鼠BBB评分降低(P <0.05),脊髓神经元皱缩畸形,呈空泡样改变;与模型组比较,葛根汤低、中、高剂量组大鼠BBB评分升高(P <0.05),且脊髓神经元损伤明显改善;(2)与正常组、假手术组比较,模型组脊髓组织中Iba-1和诱导型一氧化氮合酶蛋白表达升高(P <0.05),白细胞介素6和白细胞介素1β mRNA表达升高(P <0.05);与模型组比较,葛根汤低、中、高剂量组脊髓组织中Iba-1、诱导型一氧化氮合酶、p-NF-κB、NLRP3、ASC和cleaved Caspase-1蛋白表达以及白细胞介素6和白细胞介素1βmRNA表达降低(P <0.05);(3)与正常组、假手术组比较,模型组脊髓组织中TUNEL阳性细胞率以及Bax、Cleaved Caspase-3蛋白表达升高(P <0.05),而Bcl-2蛋白表达降低(P <0.05);与模型组比较,葛根汤低、中、高剂量组上述指标显著改善;(4)与正常组、假手术组比较,模型组大鼠脊髓组织中活性氧水平以及NOX4和p-Drp1蛋白表达升高(P <0.05),Mfn2蛋白表达降低(P <0.05);与模型组比较,葛根汤低、中、高剂量组脊髓组织中活性氧水平以及NOX4和p-Drp1蛋白表达降低(P <0.05),Mfn2蛋白表达升高(P <0.05)。结果表明,葛根汤能抑制脊髓型颈椎病模型大鼠的神经炎症反应和神经元细胞凋亡,其作用机制可能与调节NOX4/活性氧/DRP1信号通路有关。

Two New Oleanene-Type Triterpenoid Saponins from Pueraria peduncularis

From the radix of Pueraria peduncularis Grah., two new oleanene-type triterpenoid saponins named pedunsaponins B (2) and C (3) were isolated. Their structures were determined as 3-O-(6-O-methyl)-beta-D-glucuronopyranosyl-3 beta.15 alpha-diol-12-oleanene-16-one (2) and 3-O-beta-D-glucopyranosyl-( 1 --> 3)-beta-D-glucuronopyranosyl-3 beta, 15 alpha-diol-12-oleanene-16-one (3) on the basis of spectroscopic evidence and chemical reactions.

Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii

Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.

A new isoflavonolignan from Pueraria alopecuroides Craib

A new isoflavonolignan has been isolated from the Pueraria alopecuroides Craib. The structure was elucidated by the combination of 1D, 2D NMR analysis and mass spectrometry.

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.

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.

Peduncin, A New Indole Alkaloid from Pueraria peduncualris

A new indole alkaloid, peduncin was isolated from the roots of Pueraria peduncularis. Its structure was established as 1 by NMR spectroscopic and X-ray crystallographic analysis.