The mechanism of Plantaginis Semen in the treatment of diabetic nephropathy based on network pharmacology and molecular docking technology.

BACKGROUND: Diabetic nephropathy (DN) is one of the common complications of diabetes. Plantaginis Semen (PS) has a variety of therapeutic effects, however its mechanism on DN is unclear. OBJECTIVE: This paper aims to find the ingredients, the key targets, and the action pathways of PS on DN from the perspective of network pharmacology. METHOD: The databases of network pharmacology, such as Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Pharmmapper, OMIM, DrugBank, GeneCards, TTD, Disgenet, STRING, and Cytoscape software, were used to find the main ingredients and targets. Gene Ontology (GO) function and Kyoto Encyclopedia of Genome and Genomes (KEGG) pathway enrichment analysis were used to reveal the potential pathways of the PS on DN. The GEO database was used to find the targets of DN based on valid experimental research. The molecular docking technology was used to evaluate the combination between ingredients of PS and the targets. RESULTS: A total of 9 active ingredients and 216 potential therapeutic targets were obtained for PS on DN. Hub targets were discovered by the Cytoscape software analysis. CASP3 was screened by Venn diagram by making intersection between GSE30529 and hub genes. Moreover, CASP3 was combined with one of the nine active ingredients, quercetin, by molecular docking analysis. The KEGG pathways were mainly involved in diabetic nephropathy, and were simultaneously associated with CASP3 as followed: AGE-RAGE signaling pathway in diabetic complications, apoptosis, lipid and atherosclerosis, MAPK signaling pathway, TNF signaling pathway, IL-17 signaling pathway, and p53 signaling pathway. CONCLUSION: PS can have the treatment on DN through CASP3. Quercetin, as one of the nine active ingredients, can be bounded to CASP3 to inhibit apoptosis in DN. PS can also take action on DN probably through many pathways. The role of PS on DN through other pathways still needs to be further elaborated.

A Plantaginis Semen-Coptidis Rhizoma compound alleviates type 2 diabetic mellitus in mice via modulating AGEs-RAGE pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Plantaginis Semen-Coptidis Rhizoma Compound(CQC) was first recorded in Shengji Zonglu. Clinical and experimental studies have reported that both of Plantaginis Semen and Coptidis Rhizoma exerted the effects of lowering blood glocose and lipid. However, the potential mechanism of CQC on type 2 diabetes (T2DM) remain unclear. AIM OF THE STUDY: The main objective of our investigation was to explore the mechanisms of CQC on T2DM based on network pharmacology and experimental research. MATERIALS AND METHODS: Streptozotocin(STZ)/high fat diet(HFD)-induced T2DM models in mice were established to evaluate the antidiabetic effect of CQC in vivo. We obtained the chemical constituents of Plantago and Coptidis from the TCMSP database and literature sources. Potential targets of CQC were gleaned from the Swiss-Target-Prediction database, and T2DM targets were obtained from Drug-Bank, TTD, and DisGeNet. A protein-protein interaction (PPI) network was constructed in the String database. The David database was used for gene ontology (GO) and KEGG pathway enrichment analyses. We then verified the potential mechanism of CQC that were predicted by network pharmacological analysis in STZ/HFD-induced T2DM mouse model. RESULTS: Our experiments confirmed that CQC improved hyperglycemia and liver injury. We identified 21 components and gleaned 177 targets for CQC treatment of T2DM. The core component-target network included 13 compounds and 66 targets. We further demonstrated that CQC improve T2DM through various pathways, especially the AGEs/RAGE signal pathway. CONCLUSION: Our results indicated that CQC could improve the metabolic disorders of T2DM and it is a promising TCM compound for the treatment of T2DM. The potential mechanism may probably involve the regulation of the AGEs/RAGE signaling pathway.

Analysis of Chemical Composition Variations in Plantaginis Semen at Different Frying Time Based on UPLC-Q-TOF/MSE / 中国实验方剂学杂志

ObjectiveIn order to explore the changes of chemical constituents in Plantaginis Semen before and after stir-frying， ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE） was used to rapidly identify and semi-quantitatively analyze the differential components in Plantaginis Semen processed at different stir-frying time. MethodWaters ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.8 μm） was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-1 min， 5%-10%B； 1-2 min， 10%-15%B； 2-10 min， 15%-20%B； 10-12 min， 20%-40%B； 12-13 min， 40%-100%B； 13-14 min， 100%-5%B； 14-15 min， 5%B）， the flow rate was 0.3 mL·min-1， the column temperature was 40 ℃， and the injection volume was 3 μL. Electrospray ionization （ESI） was applied for mass spectrometric analysis under positive and negative ion modes， and the scanning range was m/z 50-1 500. MarkerLynx 4.1 software was used to find the differential compounds， and the intensity of each ion peak in samples with different stir-frying time was compared to study the content variations of these compounds. ResultA total of 20 components with potential significant differences were found， among which 17 were identified and 3 were unknown， mainly including phenylethanoid glycosides， iridoid glycosides， alkaloids and others. After processing， the peak intensities of 7 compounds， such as sucrose， geniposidic acid， verbascoside and plantagoguanidinic acid A， in Plantaginis Semen decreased. The peak intensities of orobanchoside， dianthoside and plantain D increased first and then decreased during the stir-frying process. The peak intensities of 10 compounds （decaffeoylacteoside， calceolarioside A， isoacteoside， etc.） increased， and 9 of them were newly generated components. ConclusionThe content and composition of the chemical components in Plantaginis Semen changed significantly after stir-frying， which may be related to the reduction of laxative effect and the enhancement of antidiarrheal and diuretic activities of Plantaginis Semen after stir-frying.

Spiky titanium dioxide nanoparticles-loaded Plantaginis Semen polysaccharide as an adjuvant to enhance immune responses.

The purpose of this study was to prepare spiky titanium dioxide nanoparticles-loaded Plantaginis Semen polysaccharide (SN-TiO2-PSP), and the structural characterization and immune response of infectious laryngotracheitis (ILT) vaccine in Hetian chickens were investigated. The structural characterization of SN-TiO2-PSP was analyzed by FT-IR, TEM, and TGA analysis. And the immune organs indexes, lymphocytes proliferation, specific antibody levels, and ratios of CD4+ and CD8+ T lymphocytes were studied. Structural characterization results showed that SN-TiO2-PSP has a typical polysaccharide absorption peak and good stability. The SN-TiO2-PSP's shape was similar to sea urchin, and its zeta potential and particle size were 27.56 mV and 976.11 nm, respectively. In vivo results showed that SN-TiO2-PSP could enhance the proliferation of peripheral lymphocytes, specific antibody levels, CD4+ and CD8+ T lymphocytes ratios, IL-4 and INF-γ levels in Hetian chickens vaccinated with ILT vaccine on D7, D14, D21, and D28. In addition, SN-TiO2-PSP not only enhanced the indexes of immune organs but also promoted the development of immune organs. Therefore, SN-TiO2-PSP has immune adjuvant activity and may become a new potential immune adjuvant.

Lipidomics study of the therapeutic mechanism of Plantaginis Semen in potassium oxonate-induced hyperuricemia rat.

BACKGROUND: Plantaginis Semen has been widely used as folk medicine and health care food against hyperuricemia (HUA) and gout, but its pharmacological mechanism remains unclear. This study investigated the therapeutic mechanism of Plantaginis Semen extract on potassium oxonate -induced HUA rats based on a lipidomics approach. METHODS: A model of HUA was established by potassium oxonate intragastric administration. 42 Sprague-Dawley (SD) male rats were randomly divided into the control group, model group, benzbromarone group (10 mg/kg) and three Plantaginis Semen groups (n = 7). The Plantaginis Semen groups were treated orally with Plantaginis Semen, 0.9375, 1.875  or 3.75 g/kg for 28 days. The levels of serum uric acid (UA), creatinine (Cr), triacylglycerol (TG) and tumor necrosis factor-α (TNF-α) were  measured using enzyme-linked immunosorbent assay kits. Ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) was used for the serum lipidomics analysis, multivariate statistical analysis and independent samples t-test were carried out for the pattern recognition and characteristic metabolites identification. The relative levels of critical regulatory factors were determined by quantitative real-time polymerase chain reaction (RT-qPCR). RESULTS: Compared with the model group, the levels of serum UA, Cr, TG and TNF-α were significantly (p < 0.05) decreased in benzbromarone and three Plantaginis Semen groups. With lipidomics analysis, significant lipid metabolic perturbations were observed in HUA rats, 13 metabolites were identified as potential biomarkers and glycerophospholipid metabolism pathway was  most affected. These perturbations  were partially restored via treatment of benzbromarone and Plantaginis Semen. Additionally, the mRNA expression levels of urate anion transporter 1 (URAT1) and phosphatidylinositol 3-kinase/protein kinases B (PI3K/Akt) were significantly decreased (p < 0.01) after treatment with benzbromarone and high dose of Plantaginis Semen. CONCLUSIONS: Plantaginis Semen had significant effects on anti-HUA, anti-inflammatory and renal protection. It attenuated potassium oxonate-induced HUA through regulation of lipid metabolism disorder.

Overview of Research on Variety，Processing and Quality Evaluation of Plantaginis Semen / 中国实验方剂学杂志

Plantaginis Semen is a traditional Chinese medicine （TCM） commonly used in China， which is one of the authentic medicinal materials in Jiangxi. It has great development prospects. However， the current research on Plantaginis Semen is not in-depth enough， mainly involving chemical components and pharmacological activities. There are few researches on processing and variety of Plantaginis Semen. In order to further develop and utilize the resources of Plantaginis Semen， we summarized 4 varieties that have been studied more at present， the processing contents of Plantaginis Semen in ancient and modern literature were consulted and sorted out， and its processing historical evolution were summarized. The influences of different processing technologies and methods on the chemical composition and pharmacological effects of Plantaginis Semen were analyzed， the possible processing mechanism was discussed. Meanwhile， and the quality evaluation methods of Plantaginis Semen varieties included in the 2020 edition of Chinese Pharmacopoeia were summarized. The author mainly analyzed the researches status of Plantaginis Semen and its decoction pieces in the three aspects of variety， processing and quality evaluation， and summarized its current major problems such as insufficient use and development of varieties， unclear processing mechanisms， and undetermined quality evaluation standards. And combined with the national standardization project of TCM to carry out the prospect and analysis for it， in order to solve the problems in the actual production and use of Plantaginis Semen， and provide reference for its further development， production of the high-quality decoction pieces， analysis of the processing mechanism， and establishment of the quality control system.

Effect of Polysaccharides from Plantaginis Semen on Renal Injury and Gut Microbiota in Rats with Membranous Nephropathy / 中国实验方剂学杂志

Objective:To study the protective effect of polysaccharides from Plantaginis Semen （PSP） against renal injury in rats with membranous nephropathy （MN） and its influence on the gut microbiota to provide a theoretical basis for the further investigation of PSP in the treatment of MN. Method:The MN model was induced by tail vein injection of cationic bovine serum albumin （C-BSA， 3.5 g·L^-1） in rats with a modeling period of seven weeks. At the 4th week of modeling， the model rats were divided into a model group， a positive drug group （benazepril hydrochloride， 10 mg·kg^-1·d^-1）， a PSP high-dose group （PSP-H， 800 mg·kg^-1·d^-1）， a PSP medium-dose group （PSP-M， 400 mg·kg^-1·d^-1）， and a PSP low-dose group （PSP-L， 200 mg·kg^-1·d^-1） according to the random number table， with 10 in each group. Ten healthy rats were assigned to the normal control group. The rats in the normal control group and the control group received an equal amount of physiological saline by gavage， and those in the groups with drug intervention were administered correspondingly，once a day，for consecutive four weeks. The pathological changes of rat kidney and colon tissues were observed by optical microscopy. The enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） and interleukin-1<italic>β </italic>（IL-1<italic>β</italic>） in the serum and colon tissues. The immunohistochemistry （IHC） was used to detect the protein expression of TNF-<italic>α </italic>and IL-1<italic>β </italic>in renal tissues. The 16S rRNA sequencing method was used to investigate the effect of PSP on the gut microbiota in MN rats. Result:Compared with the normal control group， the model group showed enlarged glomeruli， thickened basement membrane， atrophied colonic gland， increased TNF-<italic>α</italic> and IL-1<italic>β</italic> in the serum and colon tissues （<italic>P</italic><0.05）， and elevated protein expression of TNF-<italic>α</italic> and IL-1<italic>β </italic>（<italic>P</italic><0.01）. Compared with the model group， the positive drug group and the PSP-H group displayed shrunk glomerular capsules， relieved basement membrane thickening， and neatly arranged colonic mucosa in colon tissues， while the PSP-M and PSP-L groups were inferior in improving renal tissues and colon tissues. Additionally， the PSP-H and PSP-M groups showed declining TNF-<italic>α</italic> and IL-1<italic>β</italic> in the serum and colon tissues （<italic>P</italic><0.05） and dwindled protein expression of TNF-<italic>α</italic> and IL-1<italic>β </italic>in the renal tissues （<italic>P</italic><0.01）. No significant difference was observed in the PSP-L group. Compared with the normal control group， the model group showed increased abundance of Firmicutes and decreased abundance of Bacteroidetes. After PSP intervention， the abundance of Firmicutes was decreased， while that of Bacteroidetes was increased， and such changes were predominant in the PSP-H group. Conclusion:PSP can effectively alleviate renal injury， reduce the expression of inflammatory factors， regulate the structure of gut microbiota， and improve the damaged intestinal barrier of MN rats.

HPLC determination of the content for six constituents before and after salt processing of Plantaginis Semen from different habitats / 国际中医中药杂志

Objective:To compare the content change of 6 constituents in Plantaginis Semen from different habitats before and after salt processing. Methods:HPLC method was used to quantitatively analyze 6 ingredients in Plantaginis Semen and processed with salt including geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside. Results:The geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside were well separated. The linear ranges of which were 0.259 2-3.628 8 μg ( r=0.999 8), 0.054 3-0.760 5 μg ( r=0.999 6), 0.030 0-0.420 6 μg ( r=0.999 4), 0.055 6- 0.777 8 μg ( r=0.999 5), 0.287 0-4.018 0 μg ( r=0.999 8), 0.033 1-0.463 1 μg ( r=0.999 7), respectively. Average recovery rates were 98.68%, 98.46%, 98.87%, 98.99%, 98.34%, 98.75% ( n=6), respectively. There were mild differences in the contents of 6 ingredients of 8 batches of Plantaginis Semen from 5 different habitats. There were no obvious differences between the raw products and the products after salt process in Plantaginis Semen. The content of flavonoids, geniposidic acid and isoverbascoside in Plantaginis Semen were significantly increased after salt process, while the content of verbascoside was reduced. Conclusion:HPLC method to quantitatively analyze the 6 constituents in Plantaginis Semen before and after salt process could provided a reference for the quality change and the material basis for the efficacy of Plantaginis Semen before and after salt process.

Diuretic Effect and Metabolomics Analysis of Crude and Salt-Processed Plantaginis Semen.

Plantaginis Semen (PS) is well recognized in traditional Chinese medicine (TCM) and health products. Crude PS (CPS) and salt-processed CPS (SPS) are the two most commonly used decoction pieces of PS, and are included in the 2020 edition of Chinese Pharmacopoeia. Although they all have multiple effects, the mechanisms for treating diseases are different and remain unclear, the processing mechanism of SPS is also indeterminate, which hinders their clinical application to a certain extent. In order to solve these problems and further develop PS in the clinical application. Here, we used saline-loaded model rats for experiments, and utilized an integrated approach consisting of pharmacological methods and metabolomics, which could assess the diuretic impact of CPS and SPS ethanol extracts on saline-loaded rats and elucidate the underlying mechanism. The results showed that CPS and SPS both produced increased urine volume excretion and urine electrolyte excretion, but the levels of aldosterone (ALD) and aquaporin 2 (AQP2) were decreased. And 30 differential metabolites such as linoleic acid, lysoPC(O-18:0), sphingosine-1-phosphate, lysoPC(18:0) were found, mainly involving three metabolic pathways. In conclusion, CPS and SPS both have a diuretic effect, and that of SPS is better. This work investigated the possible diuretic mechanisms of CPS and SPS which may also be the mechanism of PS for anti-hypertension. In addition, a holistic approach provided novel and helpful insights into the underlying processing mechanisms of TCM.

Optimization of Processing Technology of Salt-processed Products of Plantaginis Semen Based on Response Surface Method and AHP-CRITIC Mixed Weighting Method and Investigation of Its Diuretic Effect / 中国实验方剂学杂志

Objective:To optimize the processing technology of salt-processed products of Plantaginis Semen with the specific process parameters, and verify the obtained processing technology by pharmacodynamic research, so as to provide experimental basis for the standardized production and quality control of this decoction pieces. Method:Taking composite score of appearance character score, dry extract yield and contents of three components (geniposidic acid, acteoside and isoacteoside) as index, the analytic hierarchy process (AHP)-criteria importance through intercrieria correlation (CRITIC) mixed weighting method was used to determine the weight coefficient of each index. Based on single factor tests, the response surface method was used to investigate the effects of frying time, frying temperature, salt amount and water amount on the processing technology of salt-processed products of Plantaginis Semen, and the processing technology was verified by diuretic experiment with furosemide tablets as the positive drug (administration dose of 0.01 g·kg-1). Result:The weight coefficients of geniposidic acid content, acteoside content, appearance character score, isoacteoside content and dry extract yield were 0.319, 0.193, 0.207, 0.273 and 0.008, respectively. The optimal process parameters were as following：fried at 150-180 ℃ for 10 min (obtained from the single factor tests), 100 g of Plantaginis Semen sprayed evenly with 2 g of salt (2 g of salt dissolved in 20 mL of water), and fried at 150-180 ℃ for 15 min. Compared with the blank group, both of the raw products group and the salt-processed products group could significantly increase the secretion of urine volume (P<0.01), but the excretion of Na+ in the urine of rats in the salt-processed products group was significantly higher than that in the raw products group (P<0.05). Conclusion:The optimized processing technology is simple and feasible, which can provide reference for standardizing the industrial production of salt-processed products of Plantaginis Semen. At the same time, combined with inherent quality and appearance of the salt-processed products, and verified by pharmacodynamic test, the obtained results are reasonable and reliable, which can be used for quality control of this decoction pieces.

Effect of Plantaginis Semen on Inflammatory Cytokines and mRNA and Protein Expressions of AQP4 in Colon Tissue of Diarrhea Rats / 中国实验方剂学杂志

Objective:To observe Plantaginis Semen's mechanism in treating diarrhea by observing the effect on inflammatory factors in serum and mRNA and protein expressions of aquaporin4 (AQP4) in colon tissue of diarrhea rats. Method:Senne Folium was orally administered to duplicate diarrhea rats. Sixty male SD rats were randomly divided into normal group, model group, hydrochlorothiazide group (9 mg·kg-1), and low, middle, and high-dose Plantaginis Semen groups (0.95, 1.9, 3.8 g·kg-1). Senne Folium (20 mL·kg-1) was intragastrically administered in 5 groups in the morning, except for normal group that was orally given the same dose of distilled water. In the afternoon, each treatment group was orally given the corresponding drugs, while normal group and model group were orally given the same dose of distilled water. The loose stool rate, average degree of loose stool, and diarrhea index were compared according to fecal traits and stool times after 14 days of treatment. The serum and colon tissue were collected to detect the contents of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and C-reactive protein (CRP) in serum. Hematoxylin-eosin (HE) staining was used to observe the pathological morphological changes of colon tissue, and quantiative Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expressions of AQP4 in colon tissue. Result:In the model group, the loose stool rate, average degree of loose stool, and diarrhea index were significantly increased (P<0.01), apoptosis and necrosis were observed in the epidermal cells of colonic mucosa, telangiectasia and congestion in lamina propria were obvious, and a few neutrophils were infiltrated, and the contents of TNF-α, IL-6 and CRP in serum increased (P<0.05, P<0.01), the mRNA and protein expressions of AQP4 significantly decreased (P<0.01). Compared with the model group, the loose stool rate, average degree of loose stool, and diarrhea index were significantly decreased in low, middle, and high-dose Plantaginis Semen groups (P<0.01), the apoptosis and necrosis of epidermal cells, telangiectasia and hyperemia and neutrophil infiltration in colonic mucosa were obviously improved, and the contents of TNF-α and CRP in serum significantly decreased (P<0.05, P<0.01), the mRNA and protein expressions of AQP4 increased （P<0.05, P<0.01）. Conclusion:Plantaginis Semen has a better antidiarrheal effect, and its mechanism may be related to inhibition of inflammatory reaction, repair of pathological damage of colonic mucosa, up-regulation of AQP4 expression and promotion of water and fluid metabolism.

Search of anti-allodynic compounds from Plantaginis Semen, a crude drug ingredient of Kampo formula "Goshajinkigan".

Chemotherapy-induced peripheral neuropathy (CIPN) is one of the dose-limiting side effects of cancer chemotherapy. Although the control of CIPN is important, it is difficult to manage with currently available therapeutic drugs. Therefore, there is a need for novel therapeutic agents for treating CIPN. Goshajinkigan (GJG) is a Kampo formula composed of ten crude drugs. While GJG has been used for the treatment of CIPN, the active constituents of GJG and their underlying mechanisms of pharmacological effects are still unknown. Our previous study revealed that repetitive oral administration of the water extract of Plantaginis Semen, a crude drug ingredient of GJG, inhibited the mechanical allodynia induced by an intraperitoneal injection of paclitaxel in mice. To elucidate the active compounds of Plantaginis Semen, activity-guided separation of the water extract of Plantaginis Semen was performed. From the active fraction, four iridoids (1-4) were identified. Repetitive oral administration of aucubin (1) at 100 or 30 mg/kg and 100 mg/kg of the fraction crude 3 [primarily comprised of pedicularis-lactone (3)], showed anti-allodynic activity, suggesting 1 and 3 could be some of the active compounds responsible for the anti-allodynic property of Plantaginis Semen and GJG. Our study establishes that oral administration of 1 has potent anti-allodynic effect in addition to the activity of intraperitoneally administered 1 reported previously. Identification of active anti-allodynic compounds found in Kampo formulations will support the development of novel therapies for the management of CIPN in cancer patients.

An integrative investigation on the efficacy of Plantaginis semen based on UPLC-QTOF-MS metabolomics approach in hyperlipidemic mice.

Plantaginis semen, the dried mature seed of Plantago asiatica L. or Plantago deprdssa Willd., has a prominent effect on the treatment of obesity, type 2 diabetes and lipid disorders, however, its clinical application is limited due to inadequate in-depth mechanism exploration and incomplete discussion of action targets of its in vivo. Therefore, an untargeted metabolomics approach was firstly applied to study the serum metabolic differences in mice. Metabolomics analysis was performed using ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) together with multivariate statistical data analysis. The results showed that Plantaginis semen can mainly improve blood lipids, some degree in blood glucose and insulin levels in high-fat mice, in addition, the phenotype of liver and fat stained sections demonstrated remarkable results. A total of 22 metabolites involved in arachidonic acid, glycerophospholipid, glycosphingolipid, linoleate, Omega-3 fatty acid, phosphatidylinositol phosphate and tyrosine metabolisms were identified. In further, it was found that the possible mechanisms of Plantaginis semen on hyperlipidemic mice lied in the biosynthesis of thyroxine, biological effects of enzymes of phospholipase A2 activity, glucosylceramide synthase and inositol essential enzyme 1α, genes expressions of fatty acid metabolism and inflammation. Serum metabolomics revealed that Plantaginis semen could cure the organism disease via regulating multiple metabolic pathways which will be helpful for understanding the mechanism of this herb and providing references for better applications of it in clinic, even researches on other TCMs.

Analysis of HPLC Fingerprint and Multi-component Chemical Pattern Recognition of Plantaginis Semen from Different Producing Areas in Jiangxi Province / 中国实验方剂学杂志

Objective:To establish a HPLC fingerprint detection method of Plantaginis Semen, and analyze the samples from different producing areas in Jiangxi province by combining with chemical pattern recognition method, and the contents of five ingredients in Plantaginis Semen were determined. Method:A total of 34 batches of Plantaginis Semen medicinal materials were detected by HPLC. The similarity evaluation was carried out by the 2012 edition of similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine. The chromatographic peak information was used as the data source, and three chemical pattern recognition methods were used to comprehensively analyze the quality of this medicinal herb. Quantitative analysis was performed on the 5 active components, including geniposidic acid, plantamajoside, acteoside, galuteolin and isoacteoside. Result:The similarities between Plantaginis Semen samples from different producing areas in Jiangxi province were >0.86. Orthogonal partial least squares-discriminant analysis (OPLS-DA) could distinguish samples from different producing areas, and be used to determine the chemical components, which had strong correlation with the quality of Plantaginis Semen. The contents of 5 active components in samples from different producing areas were different to some degree, especially in the content of plantamajoside. Conclusion:The established HPLC fingerprint of Plantaginis Semen has strong characteristics, combined with chemical pattern recognition method, it can effectively evaluate the quality of Plantaginis Semen and distinguish its producing areas.

Comparison of the diuretic effects of Descurainiae Semen, Coicis Semen and Plantaginis Semen / 中成药

AIM To compare the diuretic effects of Descurainiae Semen (DS),Coicis Semen (CS) and Plantaginis Semen (PS),and to observe their mechanical similarities and differences.METHODS Metabolic cage method was applied to investigating the diuretic effects of DS (2.34 g/kg),CS (7.00 g/kg) and PS (3.50 g/kg),whose diuretic mechanisms were studied by cryoscopic method,enzyme method,ion selective electrode method,ELISA and Western blot.RESULTS DS,CS and PS obviously increased saline-loaded rats' urine volume (P ＜ 0.05) and reduced their body weight (P ＜ 0.05) after administration for 7 h,which exhibited no significant effects on urine creatinine (Ucr),serum creatinine (Scr) and blood urea nitrogen (BUN)(P ＞ 0.05).DS showed its diuretic effect mainly by lowering the levels of serum Na +,atrial natriuretic peptide (ANP),brain natriuretic peptide (BNP),pulmonary AQP3,renal AQP1 and AQP2;CS showed its diuretic effect mainly by reducing the levels of serum Na +,Cl-,ANP,pulmonary AQP3,gastric AQP3,renal AQP1 and AQP2;PS showed its diuretic effect mainly by decreasing the levels of serum Na + and Cl-,pulmonary AQP3,gastric AQP3,renal AQP1 and AQP2.CONCLUSION Three medicinal materials have significant diuretic effects without obvious renal harm.DS categorized as a medicinal plant of lung channel and tropism has a great effect on netriuretic peptide system,CS categorized as a medicinal plant of spleen channel and tropism has a great effect on gastric AQP3,and PS categorized as a medicinal plant of renal channel and tropism has a great effect on renal AQPs.

Simultaneous Determination of Geniposidic Acid, Caffeic Acid, Acteoside and Isoacteoside in Plantaginis Semen Formula Granules by HPLC / 中国药学杂志

OBJECTIVE: To establish an HPLC method to simultaneously determine the contents of geniposidic acid, caffeic acid, acteoside and isoacteoside in Plantaginis Semen formula granules, in order to provide basis for studying its quality standards. METHODS: An HPLC method was developed. Kinetex C18 column (2.1 mm×100 mm, 2.6 μm)was used and eluted with mobile phase of 0.5% acetic acid-acetonitrile at the flow rate of 0.3 mL·min-1. The wavelength was set at 239 nm for geniposidic acid, 325 nm for caffeic acid, and 330 nm for acteoside and isoacteoside. The column temperature was maintained at 30℃. RESULTS: The good linear relationships between the concentration and peak area were in the range of 0.292 1-2.92 1 μg for geniposidic acid, 0.003 4-0.033 6 μg for caffeic acid, 0.047 6-0.476 μg for acteoside and 0.102 7-1.027 μg for isoacteoside (r≥0.999 8), respectively. The average recoveries were 99.32%, 98.62%, 98.23% and 98.51% with RSDs of 1.47%, 1.36%, 1.62% and 1.53%, respectively. CONCLUSION: The method is simple, feasible and reproducible and can be used for the quality control of Plantaginis Semen formula granules.

[Chemical constituents from hypoglycemic active part of Plantaginis Semen].

The chemical constituents of Plantaginis Semen with hypoglycemic effect was investigated in this paper. The previous results of the in vivo hypoglycemic effect showed that 60% ethanol extract of Plantaginis Semen decreased the levels of FBG and improved the glucose tolerance in high fat diet(HFD)-induced diabetic C57BL/6 mice. Then, in the present study, the above potential bioactive extract was separated and purified by silica gel, ODS, Sephadex LH-20 column chromatography, medium pressure liquid chromatography(MPLC)and preparative HPLC. The structures of isolated compounds were identified by physicochemical properties and spectral analyses. Eight compounds were obtained and identified as 4, 4a, 5, 7a-tetrahydro-7-(hydroxymethyl)cyclopenta[c]pyran-3(1H)-one(1), iridolactone(2), pedicularislacton(3), rehmaglutin C(4), geniposidic acid(5), p-hydroxylphenylglycerol(6), 1, 2-benzenediol-4-(2-hydroxyethyl)(7), and 3-buten-2-one-4-[3-(ß-D-glucopyranosyloxy)-4-hydroxyphenyl](8). Among them, compounds 1-5 were iridoids, and 6-8 were phenolic acids. Compound 1 was a new natural product, and compounds 2-4, 6 and 8 were isolated from the Plantaginaceae family for the first time.

Application of a quantitative 1H-NMR (1H-qNMR) method for the determination of geniposidic acid and acteoside in Plantaginis semen.

A quantitative 1H-NMR method (1H-qNMR) was developed to determine the concentration of acteoside and geniposidic acid in Plantaginis semen, which is an important crude drug for diuretic purposes. The purity of geniposidic acid and acteoside was determined by the ratio of the intensity of the H-3 signal at δ 7.51 ppm or the H-7″ signal at δ 7.58 ppm in methanol-d 4 to that of a hexamethyldisilane (HMD) signal at 0.04 ppm, respectively. The concentration of HMD was corrected with the International System of Units traceability using potassium hydrogen phthalate of certified reference material grade. The geniposidic acid content in two batches of Plantaginis semen as determined by 1H-qNMR was found to be 0.84 and 1.00 %, and the acteoside content was determined to be 0.80 and 0.93 %. We demonstrated that this method is useful for the quantitative analysis of geniposidic acid and acteoside in Plantainis semen.

Chemical constituents from hypoglycemic active part of Plantaginis Semen / 中国中药杂志

The chemical constituents of Plantaginis Semen with hypoglycemic effect was investigated in this paper. The previous results of the in vivo hypoglycemic effect showed that 60% ethanol extract of Plantaginis Semen decreased the levels of FBG and improved the glucose tolerance in high fat diet(HFD)-induced diabetic C57BL/6 mice. Then, in the present study, the above potential bioactive extract was separated and purified by silica gel, ODS, Sephadex LH-20 column chromatography, medium pressure liquid chromatography(MPLC)and preparative HPLC. The structures of isolated compounds were identified by physicochemical properties and spectral analyses. Eight compounds were obtained and identified as 4, 4a, 5, 7a-tetrahydro-7-(hydroxymethyl)cyclopenta[c]pyran-3(1H)-one(1), iridolactone(2), pedicularislacton(3), rehmaglutin C(4), geniposidic acid(5), p-hydroxylphenylglycerol(6), 1, 2-benzenediol-4-(2-hydroxyethyl)(7), and 3-buten-2-one-4-[3-(β-D-glucopyranosyloxy)-4-hydroxyphenyl](8). Among them, compounds 1-5 were iridoids, and 6-8 were phenolic acids. Compound 1 was a new natural product, and compounds 2-4, 6 and 8 were isolated from the Plantaginaceae family for the first time.

Phenols from Plantaginis Semen / 中成药

AIM To study the phenols from Plantaginis Semen.METHODS The 65％ and 95％ ethanol extracts of Plantaginis Semen were isolated and purified by macroporous resin,silica,ODS,Sephadex and preparative column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as (+)-(7R,7'R,8S,8'S)-neo-olivil (1),erythro-guaiacylglycerol-β-ferulic acid ether (2),eriodictyol (3),luteolin (4),chrysoeriol (5),hydroxytyrosol (6),4-(3,4-dihydroxyphenyl)-(E)-3-buten-2-one (7),ferulic acid (8),5,7-dihydroxychromone (9).CONCLUSION Compounds 2-3,6-7 and 9 are isolated from genus Plantago for the first time,compound 5 is first obtained from this plant.