Mechanisms of Dihuang(Rehmanniae Radix)in treating diabetic nephropathy complicated with depression based on network pharmacology

Objective To predict the molecular mechanism of Dihuang(Rehmanniae Radix)in the treatment of diabetic nephropathy(DN)complicated with depression based on network pharmacology.Methods The components of Dihuang(Rehmanniae Radix)were identified from the Integrated Pharmacology-based Research Platform of Traditional Chinese Medicine(TCMIP),Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and relevant literature.The component targets were detected by combining the SwissTargetPrediction and Pub Chem databases.Disease targets were collected from the Therapeutic Target Database(TTD),Dis Ge NET,and Ensembl databases with“diabetic nephropathy”and“depression”as keywords.The disease-component targets were mapped using Venny 2.1.0 to obtain potential targets.A protein-protein interaction(PPI)network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database and Cytoscape 3.7.2.The co-expression genes of the key targets were collected based on the COXPRESdb 7.3.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed for potential targets using R language.Target-component docking was verified and evaluated using Discovery Studio 4.5.Results According to the databases and literature reports,Dihuang(Rehmanniae Radix)contained 65 active components,and had 155 related targets for the treatment of DN complicated with depression.PPI screening showed that the key targets included serine/threonine protein kinase 1(AKT1),signal transducer and activator transcription 3(STAT3),interleukin 6(IL-6),mitogen-activated protein kinase 1(MAPK1),and vascular endothelial growth factor A(VEGFA),etc.GO enrichment analysis mainly involved biological processes,such as lipid metabolism,protein secretion regulation,cell homeostasis,and phosphatidylinositol 3 kinase activity.KEGG pathway enrichment analysis included the role of the AGE-RAGE signaling pathway in diabetic complements,insulin resistance(IR),neurotrophin signal path,Toll-like receptor signaling pathway,relaxin signaling pathway,epidermal growth factor receptor tyrosine kinase inhibitors(EGFR-TKIs),etc.Molecular docking showed that the target had high affinity for stachyose,manninotriose,verbascose,nigerose,etc.Conclusion Based on network parmacology,this study preliminarily predict the effects of Dihuang(Rehmanniae Radix)in treating DN complicated with depression by regulating inflammation,glucose metabolism,nution nerve,etc.

The dynamic changes and mechanisms of Rehmanniae radix processing based on Maillard reaction

Background:Traditional Chinese medicines are usually processed before they are used for clinical treatment.This is done in a way associated with the Maillard reaction.Methods:Based on the Maillard reaction,this paper analyzed the degree of processing of rehmannia root(Rehmanniae radix)relative to the dynamic variation rules of Maillard reaction index parameters,including pH,A420,amino acids,and 5-hydroxymethylfurfural.Furthermore,this study introduced thermal analysis techniques and pyrolysis kinetics to assess the influence of the correlation between processing raw rehmannia root and the Maillard reaction during carbonization.It then went through the whole process of transforming the raw material to end-product in order to explain the scientific connotation of processing it.Results:The results showed that each time rehmannia root was processed,its pH value and amino acid content decreased,while the A420 value and 5-HMF increased.Processing with wine shows a significant difference in these experimental indexes.The position and intensity of the maximum thermal weight loss rate peak of processed rehmannia root at different degrees of processing are different.Comprehensive quantitative 221±0.2°C for processed rehmannia root carbonization was the processing temperature limit.Moreover,the kinetic solution verified that the activation energy corresponding to the carbonization temperature was close to the maximum value of the activation energy of the whole carbonization process,and the optimal mechanism function was g(α)=((1−α)−1/3−1)2.Conclusion:The Maillard reaction occurred during the processing of rehmannia root mixed with carbonization.With each increase of the number of steaming and drying cycles involved in the processing,the level of Maillard reaction increased significantly.The wine-steaming method had a significant effect on the quality of the processed product.

Rehmanniae Radix Praeparata aqueous extract improves hepatic ischemia/reperfusion injury by restoring intracellular iron homeostasis

Hepatic ischemia/reperfusion injury(HIRI)is a common pathophysiological condition occurring during or after liver resection and transplantation,leading to hepatic viability impairment and functional deterioration.Recently,ferroptosis,a newly recognized form of programmed cell death,has been implicated in IRI.Rehmanniae Radix Praeparata(RRP),extensively used in Chinese herbal medicine for its hepatoprotective,anti-inflammatory,and antioxidant properties,presents a potential therapeutic approach.However,the mechanisms by which RRP mitigates HIRI,particularly through the regulation of ferroptosis,remain unclear.In this study,we developed a HIRI mouse model and monocrotaline(MCT)-and erastin-induced in vitro hepatocyte injury models.We conducted whole-genome transcriptome analysis to elucidate the protective effects and mechanisms of RRP on HIRI.The RRP aqueous extract was characterized by the presence of acteoside,rehmannioside D,and 5-hydroxymethylfurfural.Our results demonstrate that the RRP aqueous extract ameliorated oxidative stress,reduced intracellular iron accumulation,and attenuated HIRI-induced liver damage.Additionally,RRP significantly inhibited hepatocyte death by restoring intracellular iron homeostasis both in vivo and in vitro.Mechanistically,the RRP aqueous extract reduced intrahepatocellular iron accumulation by inhibiting ZIP14-mediated iron uptake,promoting hepcidin-and ferroportin-mediated iron efflux,and ameliorating mitochondrial iron aggregation through upregulation of Cisd1 expression.Moreover,siRNA-mediated inhibition of hamp synergistically enhanced the RRP aqueous extract's inhibitory effect on ferroptosis.In conclusion,our study elucidates the mechanisms by which RRP aqueous extracts alleviate HIRI,highlighting the restoration of iron metabolic balance.These findings position RRP as a promising candidate for clinical intervention in HIRI treatment.

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.

Deciphering the potential mechanism of Radix Rehmanniae Praeparata for treating cancer-related anemia based on network pharmacology and molecular docking technology

Background:Rehmanniae Radix Praeparata(RRP,Shu Dihuang in Cinese)is a traditional Chinese herb with multiple pharmacological effects and is commonly used to treat blood deficiency syndrome,such as cancer-related anemia(CRA),alone or in combination with other herbs.However,its main active ingredients and mechanisms of action in treating CRA remain unknown.This study aims to elucidate RRP’s potential mechanism and main active components in treating CRA by using network pharmacology and molecular docking technology system.Methods:The main components of RRP were obtained by the TCMSP database and literature search,and active components and potential targets were obtained by the SwissADME and SwissTargetPridiction databases.CRA targets were collected through GeneCards,DisGeNET,and DrugBank databases.Protein-protein interaction networks of potential targets were constructed via STRING 11.5 and analyzed visually with Cytoscape 3.9.1.The Metascape platform was used for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis,which were subsequently visualized with Cytoscape 3.9.1 or SangerBox platform.Moreover,Autodock Vina was used for the molecular docking of potential targets and main active ingredients that were visualized with PyMOL software.Results:In this study,31 main active ingredients of PPR were screened,and 155 related targets related to CRA were unearthed.Protein-protein interaction results showed that PPR’s core proteins for CRA intervention correlate to STAT3,SRC,MAPK3,MAPK1,PIK3R1,PIK3CA,and AKT.Multiple signaling pathways were closely related to the treatment of CRA intervened by PPR,including the PI3K-Akt signaling pathway,HIF-1 signaling pathway,JAK-STAT3 signaling pathway,TNF-αsignaling,cytokine signaling pathway and NF-kappB signaling pathway,which are closely involved in the proliferation and differentiation of hematopoietic stem cell and inflammatory response.Molecular docking results showed that these potential targets had good conformation with the core active components of RRP for treating CRA.Conclusion:This study revealed RRP’s main active components and potential molecular mechanisms in treating CRA,providing a reference for subsequent basic research.

Correlation between 5-hydroxymethylfurfural content and color of Rehmanniae Radix and Rehmanniae Radix Praeparata

In the present study,we explored the correlation between 5-hydroxymethylfurfural(5-HMF)and color of Rehmanniae Radix(RR)and Rehmanniae Radix Praeparata(RRP).The color was observed by human eyes under sunlight.The chromatic value and color difference were detected by a colorimeter.The water content was determined by an oven-drying method.The 5-HMF content was determined by an HPLC method.The correlation between the 5-HMF content and the color was analyzed by bivariate correlation analysis.The results showed that the color was greyish-brown and dark brown for RR,and it was black for RRP.The deepening color of RRP was reflected in the decrease of L^*,a^*and b^*values.The water content was in accord with the requirement of Chinese Pharmacopoeia.The 5-HMF contents of RR and RRP were 0.9711–25.71μg/g and 213.4–4010μg/g,respectively.The average 5-HMF contents of RR and RRP were 8.059μg/g and 1433μg/g,respectively.The 5-HMF content in RRP was higher than that in RR.The correlations between 5-HMF content and L^*,a^*and b^*values were significant.The color of RRP was deeper than that of RR.The 5-HMF content of RRP was higher than 0.02%,while it was less than 0.02%of RR,which could be used as a marker component for judging RR and RRP.

A combined quality evaluation method that integrates chemical constituents, appearance traits and origins of raw Rehmanniae Radix pieces

The quality control of Chinese herbal medicine is a current challenge for the internationalization of traditional Chinese medicine.Traditional quality evaluation methods lack quantitative analysis,while modern quality evaluation methods ignore the origins and appearance traits.Therefore,an integrated quality evaluation method is urgent in need.Raw Rehmanniae Radix(RRR)is commonly used in Chinese herbal medicine.At present,much attention has been drwan towards its quality control,which however is limited by the existing quality evaluation methods.The present study was designed to establish a comprehensive and practical method for the quality evaluation and control of RRR pieces based on its chemical constituents,appearance traits and origins.Thirty-three batches of RRR pieces were collected from six provinces,while high-performance liquid chromatography(HPLC)was applied to determine the following five constituents,including catalpol,rehmannioside A,rehmannioside D,leonuride and verbascoside in RRR pieces.Their appearance traits were quantitatively observed.Furthermore,correlation analysis,principal components analysis(PCA),cluster analysis and t-test were performed to evaluate the qualities of RRR pieces.These batches of RRR pieces were divided into three categories:samples from Henan province,samples from Shandong and Shanxi provinces,and those from other provinces.Furthermore,the chemical constituents and appearance traits of RRR pieces were significantly different from diverse origins.The combined method of chemical contituents,appearance traits and origins can distinguish RRR pieces with different qualities,which provides basic reference for the quality control of Chinese herbal medicine.

Systems response profiles to two Rehmanniae Radix formulae in metabolic syndrome patients

Objective: To explore the effects of two Rehmanniae Radix formulae in patients with metabolic syndrome(Met S), a randomized controlled study was conducted.Methods: Met S patients were randomly assigned to receive either a classic Rehmanniae Six Formula(R6, or ‘Liu Wei Di Huang Wan') or a novel multi-herbal Rehmanniae Radix containing formula SUB889 for 8 weeks. Western medicine related clinical parameters, Chinese medicine defined symptoms and syndromes as well as metabolomic profiles were evaluated at different time points.Results: R6(n = 20) and SUB889(n = 20) showed similar effects on Met S regarding the improvement of clinical parameters(waist circumference, body mass index, LDL-cholesterol, systolic blood pressure) and Qi/Yin deficiency(p < 0.05). Decreased levels of cholesteryl esters, phosphatidylcholines, triglycerides and sphingomyelins were found in the R6 group, while SUB889 formula resulted in increased levels of tricarboxylic acid cycle and glucose metabolism intermediates(malate, fumarate and pyruvate).Conclusions: R6 and SUB889 have similar effects on the treatment of Met S by improving Chinese medicine and Western medicine defined clinical outcomes. R6 is more effective in improving lipid profiles compared to SUB889. The exact mechanisms of the two formulae on Met S remain to be elucidated.

Determination and Analysis of Flavonoid Contents in Different Batches of Dried Radix Rehmannia and Prepared Radix Rehmanniae

[Objectives] This study was conducted to understand Radix Rehmannia quality and provide reference for selection in application. [Methods] The contents of flavonoids in five batches of commercially available samples were determined by ultraviolet spectrophotometry, and the precision and reproducibility of the determination method were analyzed. [Results] The contents of flavonoids in different batches of Radix Rehmannia samples were not completely the same, but the differences were not large overall. The experiment results have good reliability. As a medicinal plant, the climate and soil conditions of the producing areas would have an impact on the quality of Radix Rehmannia. [Conclusions] The contents of flavonoids determined in Radix Rehmannia were relatively stable, the quality was basically the same, and the overall quality was good.

Jiaohong pills attenuate neuroinflammation and amyloid-βprotein-induced cognitive deficits by modulating the mitogen-activated protein kinase/nuclear factor kappa-B pathway

Background:Jiaohong pills(JHP)consist of Pericarpium Zanthoxyli(PZ)and Radix Rehmanniae,two herbs that have been extensively investigated over many years due to their potential protective effects against cognitive decline and memory impairment.However,the precise mechanisms underlying the beneficial effects remain elusive.Here,research studies were conducted to investigate and validate the therapeutic effects of JHP on Alzheimer's disease.Methods:BV-2 cell inflammation was induced by lipopolysaccharide.AD mice were administered amyloid-β(Aβ).Behavioral experiments were used to evaluate learning and memory ability.The levels of nitric oxide(NO),tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)were detected using enzymelinked immunosorbent assay(ELISA).The protein expressions of inducible nitric oxide synthase(iNOS)and the phosphorylation level of mitogen-activated protein kinase(MAPK)and nuclear factor kappa-B(NF-κB)were detected using Western blot.Nissl staining was used to detect neuronal degeneration.Results:The results demonstrated that an alcoholic extract of PZ significantly decreased the levels of NO,IL-1β,TNF-α,and iNOS;increased the expression level of IL-10;and significantly decreased the phosphorylation levels of MAPK and NF-κB.These inhibitory effects were further confirmed in the AD mouse model.Meanwhile,JHP improved learning and memory function in AD mice,reduced neuronal damage,and enriched the Nissl bodies in the hippocampus.Moreover,IL-1βand TNF-αin the cortex were significantly downregulated after JHP administration,whereas IL-10showed increased expression.Conclusions:It was found that JHP reduced neuroinflammatory response in AD mice by targeting the MAPK/NF-κB signaling pathway.

地黄炮制过程中HPLC-RID指纹图谱及模式识别研究

为建立地黄(Rehmanniae radix)不同炮制品炮制过程中HPLC-RID特征图谱及多种糖含量测定方法,结合模式识别方法分析炮制过程中糖类成分变化趋势,采用Agilent ZORBAX NH2色谱柱(250.0mm×4.6 mm,5μm),以乙腈-纯水(70∶30)混合溶液为流动相,体积流量1.0 mL/min,柱温40℃,流通池温度50℃,建立地黄不同炮制品HPLC-RID特征图谱分析方法,外标法测定地黄不同炮制品中果糖、葡萄糖、蔗糖、棉子糖、水苏糖的含量;借助主成分分析(PCA)和聚类分析(HCA)模式识别方法分析鲜地黄-生地黄-熟地黄糖成分的差异与关联。结果表明,分析了12批地黄样品,匹配出HPLC-RID特征图谱共有峰6个,指认其中5个色谱峰,地黄片和鲜地黄无显著差异;鲜地黄中水苏糖含量最高;鲜地黄-生地黄-熟地黄炮制过程中,水苏糖含量锐减,果糖含量一直递增;PCA和HCA能较好地区分地黄不同炮制品,且结果一致。HPLC-RID特征图谱能反映地黄炮制过程中的变化,HPLC-RID法同时测定地黄中5种糖含量简便可靠,为地黄糖类成分提取工艺筛选及产品开发奠定研究基础。

Study on Trace Elements in Rehmannia glutinosa Libosch. by Principal Component Analysis and Clustering Analysis

[Objective] This study aimed to investigate the trace elements in Rehman- nia glutinosa Libosch. by using principal component analysis and clustering analysis. [Method] Principal component analysis and clustering analysis of R. glutinosa medicinal materials from different sources were conducted with contents of six trace elements as indices. [Result] The principal component analysis could comprehen- sively evaluate the quality of R. glutinosa samples with objective results which was consistent with the results of clustering analysis. [Conclusion] Principal component analysis and clustering analysis methods can be used for the quality evaluation of Chinese medicinal materials with multiple indices.

The marriage of sealant agent between structure transformable silk fibroin and traditional Chinese medicine for faster skin repair

Fast skin repair is critical for less infection, less pain and high quality of life, which is still limited with undesirable rehabilitation speed and side effects. Currently, laser-activated silk sealant agent without suture and gauze has been demonstrated promising for fast skin repair taking advantage of its structural transformation after heating. Nevertheless, more efficient healing effects and less side effects of laseractivated silk sealant agent remains challenging due to absence of suitable photo-thermal materials and robust/biomimetic protein materials. In this work, the marriage between silk protein and Rehmanniae radix preparata(a kind of the traditional Chinese herb) has been demonstrated as a novel and effective way to achieve an excellent healing effect for skin repair. The non-toxicity, high photothermal conversion efficiency and healing mechanism are systematically studied and proved. This new methodology might shed a new light for combining dark traditional Chinese medicine and silk fibroin for advanced wound healing technology.