Optimization of Enzymatic Extraction Process of Polysaccharides from Pseudostellaria heterophylla Fibrous Roots by Response Surface Methodology and Its Pilot Application

[Objectives]To study and optimize the process conditions of enzymatic hydrolysis technology for extracting polysaccharides from Pseudostellaria heterophylla fibrous roots and its application in workshop pilot tests.[Methods]P.heterophylla fibrous roots were taken as the matrix material,and Box Behnken design was used to analyze the extraction time,composite enzyme addition amount,and liquid-solid ratio for response surface optimization experiments,and then applied to the pilot extraction of P.heterophylla fibrous roots.[Results]Response surface analysis showed that all factors had a significant impact on the experimental indicators.The optimal extraction process conditions for polysaccharides from P.heterophylla fibrous roots were extraction time of 2.7 h,compound enzyme addition of 2.5%,and liquid-solid ratio of 32.The yield of polysaccharides from P.heterophylla fibrous roots was 4.83%.The water extraction process of P.heterophylla fibrous roots extraction pilot was used as the control group for response surface optimization of the pilot experiment.The optimization results showed that the extraction time was 3 h,the amount of composite enzyme added was 2.5%,and the liquid-solid ratio was 28.The polysaccharide yield was 4.75%,an increase of 4.63%compared to the control group.[Conclusions]This paper could provide feasibility for the innovation of enzy-matic hydrolysis technology for P.heterophylla fibrous roots and its workshop pilot practice application,as well as a reference for the industrial application of its medicinal resources.

Study on Pathogen and Bio-control of Dampingoff Disease in Pseudostellaria heterophylla in Qiandongnan Prefecture of Guizhou Province

Fungal diseases occur seriously in Pseudostellaria heterophylla in Qian- dongnan Prefecture of Guizhou Province from March to May every year. By field in- vestigation, tissue isolation and microscopic identification, the main pathogen was iden- tified as Rhizoctonia solanL the pathogen of damping-off disease. At the same time, Alternaria alternate was also isolated from the sampled diseased soil, and it has pathogenicity to Pseudostellaria heterophylla seedlings. The bio-control test showed that GH18 strain （Bacillus spp.） had significant inhibitory effects on Rhizoctonia solani and Alternaria alternate.

太子参(PSEUDOSTELLARIA HETEROPHYLLA)病毒病病原鉴定

太子参(Pseudostellaria heterophylla)是我国传统的中药材之一,其病毒病十分严重,田间发病率和种根带毒率可达100％,平均90％左右。自太子参病株分离得到的T-17、T-32、T-69和T-01四个病毒分离物,经鉴定,它们分别为芜菁花叶病毒(Turnip mosaicvirus,TuMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、蚕豆萎蔫病毒(Broad beanwilt virus,BBWV)和烟草花叶病毒(Tobacco mosaic virus,TMV)。其中以TuMV分布最广,为害最重,是太子参病毒病的主要病原。

HETEROPHYLLIN-A AND B,TWO CYCLOPEPTIDES,FROM THE ROOTS OF PSEUDOSTELLARIA HETEROPHYLLA

Two cyclopeptides,heterophyllin A and B,have been isolated from the roots of Pseudostellaria heterophylla.Their structures were elucidated by chemical, spectroscopic,and enzymatic methods.

Effects of Different Extract of Pseudostellaria Heterophylla on Immunological Function in Mice based on Meta-analysis and Network Meta-analysis

To evaluate the effects of different extract of Pseudostellaria heterophylla on immunological function in mice based on Meta-analysis and Network meta-analysis,the article retrieved domestic and foreign databases according to the"PICO"retrieval strategy,and used Stata and ADDIS software for Meta analysis.A total of 6 reports,10 randomised controlled trials(RCTs)were eventually involved.Meta analysis results showed that:compared with the control group,the experimental group of polysaccharide,saponin and crude extract were better than that of the control(P<0.05),which significantly improved the immunological function in mice.Network meta-analysis results showed that the saponin had the best effect on the increase of phagocytic index and the differences were statistically significant[MD=1.50,95%CI(0.71,2.28),P<0.05];The polysaccharide and saponin had better effect on the increase of spleen index than the control and crude extract,and the differences were statistically significant[MD=0.77,95%CI(0.27,1.31),P<0.05],[MD=0.71,95%CI(0.01,1.50),P<0.05];the polysaccharide had the best effect on the increase of thymus index and the differences were statistically significant[MD=0.70,95%CI(0.11,1.34),P<0.05].The rank probability showed that the saponin of Pseudostellaria heterophylla had the maximum probability to increase phagocytic index of mice;the probability for the components of Pseudostellaria heterophylla increasing spleen index of mice was in the order of crude extract>polysaccharide>saponin;the probability for the components of Pseudostellaria heterophylla increasing thymus index of mice was in the order of saponin>polysaccharide>crude extract.Based on the available evidence,the extract of Pseudostellaria heterophylla could improve the immunity of mice,and the clinical effect of polysaccharide and saponin was the best,which provided a more valuable scientific reference for evidencing that the polysaccharide and saponin of Pseudostellaria heterophylla was the effective components for improving immunological function,and also was conducive to the proper further development of Pseudostellaria heterophylla resources.

Optimization of Steam Explosion Process Condition for Extracting Polysaccharides from Pseudostellaria heterophylla by Response Surface Methodology

【Objective】To optimize the steam explosion process condition for extracting polysaccharides from Pseudostellaria heterophylla.【Method】The effects of steam pressure,pressure-maintaining time and material moisture content on the extraction of polysaccharides from Pseudostellaria heterophylla were studied by response surface methodology based on Box-Behnken design.【Result】The findings showed that each factor could significantly affect the test index,and the optimum condition was as follows:steam pressure 1.50 Mpa,pressuremaintaining time 46 s and material moisture content 46%.Under this condition,the verified experimental value of polysaccharides from Pseudostellaria heterophylla was 39.32%,indicating a relative standard deviation of 2.73%from the predictive value.Meanwhile,scanning electron microcopy(SEM)images showed that the surface physical structure of Pseudostellaria heterophylla was irregularly broken and cracked,which means the physical structure of Pseudostellaria heterophylla was changed and destroyed at the cellular level.【Conclusion】This experiment provides a new approach for the extraction of polysaccharides from Pseudostellaria heterophylla,as well as a reference for the resource utilization of Pseudostellaria heterophylla.

Investigation and Green Prevention and Control Strategies of Pseudostellaria heterophylla Diseases in Genuine Producing Area in Zherong County

Zherong County of Fujian Province is a genuine producing area of Pseudostellaria heterophylla,and disease has become a main factor restricting the healthy development of P.heterophylla industry in the county.In order to scientifically understand the occurrence status of agricultural diseases and serve the GAP of P.heterophylla,P.heterophylla diseases in Zherong County were investigated,and the information and regularity of diseases were summarized.P.heterophylla diseases in Zherong County were infectious,and caused by fungi and viruses.The diseases occurred frequently in spring and summer,and symptoms were manifested in pathological changes of leaf and tuber.Eventually,green prevention and control strategies of diseases were put forward:cultivating new disease-resistant varieties,scientifically administrating drugs according to symptoms,administrating drugs in optimum period,strengthening biological control,strictly requiring drug safety period and paying attention to green prevention and control during the planting process.The occurrence of P.heterophylla diseases in Zherong County was investigated and summarized,and the corresponding strategies were put forward,so as to scientifically serve the agricultural industry in P.heterophylla genuine producing area,and to provide a basis for the sustainable development of modern pollution-free planting industry of P.heterophylla.

Effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on Sexual Maturity of SD Female Rats

[Objectives] To study the effects of Panax ginseng,Codonopsis pilosula and Pseudostellaria heterophylla on the growth and development of SD female rats. [Methods] The rats were randomly divided into control group,P. ginseng group,C. pilosula group,and P. heterophylla group,intragastrically administered with physiological saline,P. ginseng,C. pilosula,and P. heterophylla for 2 weeks,daily vaginal smear inspection was performed,estradiol( E_2) level before and after intervention was detected,rats were dissected,took out ovary and uterus,and calculated the ovarian index and uterus index. [Results]In 4 groups of rats,there was no significant difference in the time of sexual maturity and ovarian index; compared with the control group,E_2 level of C. pilosula group and P. ginseng group were higher( P < 0. 05);compared with the control group and P. heterophylla group,the ovarian index of P. ginseng group was higher( P < 0. 05); compared with the control group,the body weight of three administration groups increased much more( P < 0. 05). [Conclusions] P. ginseng and C. pilosula may lead to sexual maturity of SD female rats,while P. ginseng,C. pilosula and P. heterophylla can promote increase of body weight of SD female rats.

Saponins Content in Fibrous Roots of Pseudostellaria heterophylla and Optimization of the SEP Extraction Technology

[Objectives]To explore the saponins resources in fibrous roots of Pseudostellaria heterophylla and optimize the SEP technology for extracting saponins.[Methods]Saponins contents in fibrous roots and root tubers of P.heterophylla were compared;with the fibrous roots as host materials,this experiment applied Box-Behnken design to analyze the influence of steam pressure,maintenance pressure time,moisture content of host materials on the content of saponins extracts.[Results]Saponins content was higher in fibrous roots than in tuberous roots of P.heterophylla;the response surface optimization showed that SEP(steam explosion pretreatment)promoted the contents of saponins and extracts in fibrous roots,and the optimal extracting conditions were stream pressure 1.50 Mpa,maintenance pressure time 54 s,and moisture content of host materials 30%.Under this extracting conditions,the verification value of saponins content experiment was 0.441%,36.53%higher than the untreated control group.[Conclusions]This research offered a new method of exploring saponins resources from fibrous roots of P.heterophylla,and provided a reference for developing new medicinal parts of P.heterophylla.

Pseudostellaria heterophylla:Antioxidant Activity and Its Mechanism of Action

The tonic traditional Chinese medicine Pseudostellaria heterophylla both medicinal and nutritional effects.It has been applied to research in the field of modern aquaculture,and its anti-oxidant clinical trials have also become a hot spot.Through searching domestic and foreign literature databases to sort out related literature,the anti-oxidation property of P.heterophylla was reviewed systematically from the three levels of in vitro chemical method,biological method and animal experiment method.On this basis,the functional mechanism of the anti-oxidant activity of P.heterophylla explored,in order to provide a reference for the follow-up in-depth research on the anti-oxidation of P.heterophylla and the discovery and development of feed and health care value of P.heterophylla the field of modern animal breeding.

In Vitro Probiotic Effect of Pseudostellaria heterophylla Fibrous Roots on Feed Bacillus subtilis and Preliminary Study on Its Solid Fermentation

Traditional Chinese medicine Pseudostellaria heterophylla fibrous roots have been widely studied and applied in non-antibiotic clinical breeding of livestock and poultry. In vitro probiotic effect of P. heterophylla fibrous roots extract on feed probiotics Bacillus subtilis BD-K010 was studied, and the feasibility of solid fermentation of P. heterophylla fibrous roots was preliminarily evaluated. For in vitro probiotic effect, the increased concentration of P. heterophylla fibrous roots extract dependently increased the total biomass of B. subtilis BD-K010;1.0% P. heterophylla fibrous roots extract received the best effect, and the final p H of 1.0% experimental group was closer to neutral. Meantime, B. subtilis BD-K010 with optimum concentration of P. heterophylla fibrous roots showed significantly higher in vitro antibacterial effect than the control group(P<0.01),and the antibacterial effects on Escherichia coli and Staphylococcus aureus were improved by 51.99% and 63.16%, but it was ineffective to Salmonella. For solid fermentation, the profile of substrate complex and appendage flocculent structure on substrate surface at the end of fermenta-tion in experimental groups added with B. subtilis BD-K010 and cellulase plus BD-K010 were more complex;the live bacteria number, polysaccharide content and saponin content at fermentation end-point in two experimental groups were extremely higher than those in the control group(P<0.01).P. heterophylla fibrous roots extract had good in vitro probiotic effect on B. subtilis BD-K010 and promoted its antibacterial effect, and it is feasible to use probiotics for solid fermentation of P. heterophylla fibrous roots and to improve effective components. It is of great significance to further de-velop and utilize P. heterophylla fibrous roots resources in modern animal husbandry.

Key Points for High-yield Cultivation Management of Pseudostellaria heterophylla in Weihai Area

[Objectives] To explore the scientific cultivation mode of Pseudostellaria heterophylla in Shandong Province. [Methods] According to the ecological characteristics of P. heterophylla, the cultivation techniques and management points of P. heterophylla in Weihai are summarized. [Results] The scientific cultivation techniques and management points of P. heterophylla are introduced from the aspects of land selection, land preparation, seed selection, seedling nursing, timely sowing and transplanting, field management and disease and pest control. [Conclusions] This study will provide theoretical guidance for the high-quality, high-yield and high-efficiency cultivation of P. heterophylla in Shandong area.

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China

Objective:Pseudostellaria heterophylla has been paid more attention in recent years,mainly as a medicine food homology plant.The content determination of P.heterophylla is not specified in the Chinese Pharmacopoeia(version 2020).The environmental conditions in different production areas could exert an influence on the quality of P.heterophylla.The purpose of this study is to discriminate P.heterophylla collected from different geographical origins of China.Methods:In this study,the content of polysaccharide in 28 batches of P.heterophylla was determined using phenol–sulfuric acid.HPLC fingerprints were established under optimised HPLC-PDA methods.Subsequently,the similarity analysis(SA)and the quantification of heterophyllin B were analyzed.The metabolites of P.heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system.Principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),hierarchical cluster analysis(HCA)and orthogonal PLS-DA(OPLS-DA)were performed based on all peak areas.Results:The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas,which varied significant from different origins.While the content of heterophyllin B in Anhui and Jiangsu was high.The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990,and the characteristic map can be used to identify and evaluate the quality of P.heterophylla.The samples from Fujian,Guizhou,Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA,PLS-DA,HCA,OPLS-DA,indicating that their metabolic compositions were significantly different.Ultimately,a total of 15 metabolites which were filtrated by a VIPvalue>1 and a P-value<0.05 associated with the separation of different origins were identified.Conclusion:HPLC fingerprint was established to evaluate the quality and authenticity of P.heterophylla.The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions.A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P.heterophylla from different origins in China for the first time.Overall,this study provides insights to metabolomics of P.heterophylla and supplies important reference values for the development of functional foods.

Alleviatory effect of rare earth micro-fertilizer on photosystemⅡ(PSⅡ)photoinhibition in Pseudostellaria heterophylla leaves at photosynthetic midday depression

In order to explo re the mitigatory mechanism of rare earths on the midday depression of photosynthesis,this study used fast-phase chlorophyll fluorescence technology combined with an antioxidant enzyme system to analyze the effects of lanthanum and cerium on the photosystemⅡ(PSⅡ)function of Pseudostellaria heterophylla leaves at noon.The results show that the maximum photochemical efficiency(F_(v)/F_(m))and the photosynthetic performance index(PI_(ABS))of P.heterophylla leaves during the photo synthetic noon-break are only 0.73 and 0.86,respectively.The leaves of P.heterophylla show obvious photoinhibition.The application of two rare earth fertilizers promotes the light energy absorption,capture,conversion,and electron transfer efficiencies of PSⅡin the P.heterophylla leaves and alleviates the photoinhibition caused by excess excitation energy.Additionally,the effect of lanthanum is better than that of cerium.In the fertilization groups,P.heterophylla leaves show higher PSⅡphotochemical activity.Fertilization protects the oxygen-evolving complex(OEC)activity of P.heterophylla,thus reducing oxidative damage to the chloroplasts and preventing the dissociation of the thylakoids to maintain the normal physiological functioning of PSⅡ,thereby reducing the degree of photoinhibition.Additionally,fertilization increases the electron transfer ability from Q_(A) to Q_(B) on the PSⅡelectron receptor side of P.heterophylla.Correlation analysis shows that the fluorescence parameters V_(L),V_(K),and V_(J) are significantly negatively correlated with antioxidant enzyme activity and are significantly positively correlated with malondialdehyde content,relative electric conductivity and reactive oxygen species(ROS),which further indicates that rare earths can enhance the ROS scavenging system in plants and reduce the degree of membrane lipid peroxidation and plasma membrane damage,thereby maintaining the structure and function of the OEC on the donor side of PSⅡ,increasing the electron receptor pool,and contributing to the defense against photoinhibition.

太子参脱毒苗培养、化学成分及指纹图谱研究进展

太子参是我国的传统中药材,含有多种生物活性成分,如糖类、环肽类、皂苷类、微量元素、挥发油类等。现代药理学研究表明这些化学成分具有增强记忆、免疫调节、抑制食管癌、降血糖、促血管新生等活性。近年来,多地大力推广人工栽培,建设太子参GAP基地。但亦出现了一些问题,如太子参病毒病呈逐年加重、品种混乱、内在质量参差不齐等。针对这些问题学者们开展了诸多研究。本文从太子参脱毒苗研究、化学成分研究及指纹图谱研究等方面进行综述,以期为该资源的种植栽培、药物研发、质量标准建立等提供参考。

樟芝-太子参双向液体发酵培养基优化及效应分析

【目的】有效利用太子参根须副产物资源,提高樟芝(Antrodia camphorata)菌丝产量,促进活性成分合成。【方法】以太子参根须(粉末)作为培养基组分,Plackett-Burman试验筛选影响樟芝菌丝生长的关键成分,正交试验优化培养基组成。【结果】筛选试验结果表明,葡萄糖、酵母粉、蛋白胨及KH2PO4对樟芝菌丝生长有重要影响,正交试验优化的培养基组成为:太子参6 g·L^(−1),葡萄糖10 g·L^(−1),酵母粉4 g·L^(−1),蛋白胨6 g·L^(−1),KH_(2)PO_(4)1.5g·L^(−1)。利用优化培养基深层发酵,樟芝生物量达到4.73 g·L^(−1),与对照培养基及PDB培养基相比,胞内三萜含量分别提高2.75%和24.85%,胞外多糖分别提高161.11%和113.64%;发酵液中高密度无性孢子含量达1.8×10^(7)个·mL^(-1)。【结论】太子参适合作为樟芝双向液体发酵的药性成分,可有效促进樟芝菌丝生长、活性成分合成和无性孢子形成,是太子参根须副产物资源利用新途径。

太子参多糖对OVA免疫小鼠肠道免疫应答能力的影响

【目的】研究太子参多糖(PHPS)对卵清白蛋白(OVA)免疫小鼠肠道免疫应答的影响,为了解PHPS能否增强小鼠机体的免疫应答能力提供依据。【方法】将72只健康清洁级(SPF)雌性ICR小鼠随机分成空白对照组(CK组)、OVA免疫对照组(OVA组)、铝佐剂阳性对照组(AL组)及PHPS各剂量组[50 mg·kg^(-1) PHPS(PHPS-L)组、100 mg·kg^(-1) PHPS(PHPS-M)组、200 mg·kg^(-1) PHPS(PHPS-H)组],每组12只小鼠。试验第1~5天和第16~20天,PHPS各剂量组小鼠灌胃对应剂量的PHPS,其余各组均灌胃双蒸水;试验第6天和第21天,CK组小鼠按0.2 mL·只^(-1)的剂量腹股沟皮下注射无菌PBS,AL组小鼠按0.2 mL·只^(-1)的剂量腹股沟皮下注射OVA与铝佐剂的混合液,其他组小鼠均按0.2 mL·只^(-1)的剂量腹股沟皮下注射OVA溶液;试验第36天采用颈椎脱臼法处死小鼠,采集样品,制作十二指肠、空肠、回肠切片检测小鼠肠道形态结构和上皮内淋巴细胞数量,检测小鼠肠道细胞因子(IL-6、TNF-α)和分泌性免疫球蛋白A(SIgA)的含量,以及IL-6、TNF-αmRNA的转录水平。【结果】(1)与OVA组相比,PHPS各剂量组小鼠十二指肠、空肠、回肠的绒毛高度和绒隐比提高,PHPS-M组十二指肠、PHPS-H组空肠、PHPS各剂量组回肠的隐窝深度显著降低(P<0.05);同时,PHPS各剂量组十二指肠、空肠、回肠的上皮内淋巴细胞数量均有所增加。(2)与OVA组相比,PHPS各剂量组小鼠十二指肠、空肠、回肠IL-6、TNF-α的含量增加,SIgA含量显著增加(P<0.05)。(3)与OVA组相比,PHPS各剂量组小鼠十二指肠、空肠、回肠IL-6、TNF-αmRNA的转录水平显著上调(P<0.05)。【结论】PHPS能在一定程度上改善肠道形态结构,促进肠道上皮内淋巴细胞分泌,调节相关免疫分子的释放与表达,从而增强肠道黏膜免疫反应,提高小鼠对OVA的免疫应答能力。

太子参化学成分及生物活性研究进展

太子参为石竹科植物孩儿参的干燥块根,味甘、微苦,性平,归脾肺经,具有益气健脾、生津润肺之功效,常用于气虚津伤的肺虚燥咳,被国家卫生健康委员会批准用于保健食品。现代药理和临床研究表明,太子参中特征性化学成分多糖和环肽具有免疫调节、抗炎、改善记忆等作用。近年来对太子参的化学成分、结构和药理作用的综述仅局限于特征成分,对其他成分还鲜有系统梳理报道。因此本文系统梳理了太子参的化学成分及生物活性研究进展,以期为太子参的开发利用提供参考和依据。

不同遮光处理对太子参苗期生长及生理特性的影响

为探究遮光对太子参苗期生长及生理特性的影响,对太子参进行30%、50%的遮光和全光照处理,测定不同遮光处理对太子参苗期生长及部分生理指标的影响。结果表明,随着遮光强度的增加,太子参的叶片数、分枝数、子根数、根长、生物量等生长指标均表现为先上升后下降,30%遮光处理的太子参生长最好。遮光可显著降低太子参叶片的酶活性和丙二醛含量,且均以30%遮光处理的含量最低;在30%遮光处理下太子参叶片的SOD、POD、CAT、APX酶活性和丙二醛的含量分别为129.74 U·g^(-1)、789.32 U·(g·min)^(-1)、34.82 U·(g·min)^(-1)、211.77 U·(g·min)^(-1)FW、0.06μmol·g^(-1),比对照分别降低了35.52%、31.04%、13.73%、23.77%、45.45%。30%遮光处理的叶片可溶性糖含量(69.53 mg·g^(-1))和可溶性蛋白含量(4.27 mg·g^(-1))均为最高。2种遮光处理的叶片叶绿素a和叶绿素b含量差异不显著(P>0.05),但显著高于全光照处理(P<0.05);30%遮光处理的叶片类胡萝卜素含量为0.12 mg·g^(-1),显著高于全光照处理(P<0.05)。在人工栽培过程中,遮光度控制在30%左右对太子参苗期的生长及生理最有利。

植物乳杆菌发酵添加太子参的培养基优化及抗氧化活性研究

目的:优化植物乳杆菌(Lactiplantibacillus plantarum)发酵太子参的培养基,为太子参资源的利用和产品开发提供一定科学依据。方法:以菌体密度为指标,通过Plackett-Burman试验筛选对菌体细胞生长有重要影响的培养基组分,通过正交试验确定响应面试验的中心点,通过Box-Behnken响应面试验优化培养基组成;利用优化培养基发酵并测定发酵液(冻干粉)的抗氧化活性。结果:太子参水提物适合作为植物乳杆菌发酵的培养基成分;响应面法优化的培养基为麦芽糖29.7 g·L^(-1)、酵母粉27.5 g·L^(-1)、太子参水提物7.6 g·L^(-1)和MgSO40.5 g·L^(-1),优化条件下菌体密度达到1.11×10^(8)CFU·mL^(-1);优化培养基发酵液DPPH自由基清除率高于未发酵培养基(对照),低于MRS培养基发酵液(参照),ABTS+自由基清除率和FRAP值高于对照和参照。结论:太子参水提物可作为植物乳杆菌发酵的优良培养基成分,提高菌体密度;植物乳杆菌发酵可提高抗氧化活性。