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Objective: To investigate the targets and mechanism of Achyranthis bidentatae radix and Morindae officinalis radix (ABR-MOR) for the treatment of osteoporosis (OP) by utilizing network pharmacology, molecular docking technology (MDT) and molecular dynamics simulation (MDS). Methods: The main drug active ingredients (DAIs) and target genes of ABR-MOR were screened by the TCMSP database. The relevant targets of OP were obtained from GeneCards, DisGeNET, and CTD databases. Venny mapping is used to determine the potential target of ABR-MOR in the treatment of OP. The potential targets were analyzed using a proteinâprotein interaction network and the MCODE module, and were subjected to GO and KEGG enrichment analysis. The binding sites and conditions of potential key DAIs and core targets were verified through MDT and MDS. Result: The 32 DAIs and 212 targets of ABR-MOR were screened; 1453 OP-related targets were obtained, and 118 targets were mapped. The results of GO and KEGG enrichment analysis showed that the targets of DAIs-OP were mainly enriched in biological processes such as response to hormones, peptides, oxygen levels and reactive oxygen species, and positive regulation of cell migration. The main signaling pathways enriched in the regulation of the immune-inflammatory response, cell proliferation, senescence, apoptosis, angiogenesis, and estrogen signaling pathway. Additionally, the targets were also enriched in bone metabolism-related cell differentiation biological processes and the osteoclast differentiation signaling pathway. MDT and MDS results showed that wogonin, beta-sitosterol, and americanin A, the core DAIs of ABR-MOR, were able to form good ligandâprotein complexes with key targets such as PTGS2, PTGS1, PRKACA, PGR, MAPK1, AKT1, and RELA. Conclusion: This study preliminarily investigated the key targets, biological processes, and signaling pathways involved in the combined application of ABR and MOR for the treatment of OP. The results revealed that ABR-MOR may play a therapeutic role mainly by regulating immune-inflammatory responses, cellular biological processes, and osteoblast differentiation, which provides a theoretical basis for further experimental validation and a new strategy for the treatment of OP.
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In this study, the hot water extraction process of crude polysaccharides from Morindae officinalis radix (cMORP) was conducted and optimized through a single-factor test and orthogonal experimental design. With the optimal extraction process (extraction temperature of 80 °C, extraction time of 2 h, liquid/solid ratio of 15 mL/g, and number of extraction of 1), the cMORP was obtained by the ethanol precipitation method. The chemical properties and preliminary characterization of the cMORP were analyzed by chemical or instrumental methods. Furthermore, to indicate a preliminary study on safety, a single oral dose of 5000 mg/kg body weight (BW) was administered orally to Kunming (KM) mice for acute toxicity, and the cMORP was administered orally to KM mice once a day at doses of 25, 50, and 100 mg/kg BW for 30 days. General behaviors, body weight variations, histopathology, relative organ weights, and hematological and serum biochemical parameters were observed and recorded. The results suggested there were no toxicologically significant changes. Based on the safety study, cMORP can be initially considered non-toxic with no acute oral toxicity up to 5000 mg/kg BW and safe at up to 100 mg/kg BW in KM mice for 30 days.
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Morindae Officinalis Radix (MOR) and Alpiniae Oxyphyllae Fructus (AOF) have been widely used as dietary supplements and traditional herbal medicines for centuries. Fungal and mycotoxin contamination in MOR and AOF has been reported recently. In this study, fungi in MOR and AOF are first investigated using DNA metabarcoding, and the differences in fungal microbiome between moldy and non-moldy samples are analyzed. The results show that Ascomycota is the most prevailing fungus at the phylum level in MOR and AOF with relative abundances of 49.53-94.32% and 14.81-81.85%, respectively. Penicillium (1.86-76.14%), Cladosporium (1.82-56.65%), and Trichoderma (0.12-19.71%) are the dominant genera in MOR. Penicillium (0.27-56.06%), Papiliotrema (0.04-51.71%), and Cladosporium (3.08-44.41%) are the dominant genera in AOF. Two potential toxigenic fungi were detected, namely, Trichoderma atroviride and Fusarium equiseti. Moreover, the differences in fungal communities between moldy and non-moldy samples were monitored. In conclusion, DNA metabarcoding can be used to assess the fungal microbiome in edible medicinal herbs, thereby providing a basis for ensuring food safety and drug efficacy.
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By consulting ancient herbal medicines， medical and prescription books， combined with modern documents， the textual research of Morindae Officinalis Radix has been conducted to verify the name， origin， changes in production areas， quality evaluation， harvesting， and processing methods， so as to provide reference and basis for the development and utilization of the famous classical formulas. After textual research， the production areas of Morindae Officinalis Radix has experienced great changes from north to south in history. The original plants involve 11 families， 14 genera and 21 species， and the mainstream varieties in ancient times were Damnacanthus officinarum and D. indicus， and the basis of Morindae Officinalis Radix in modern times has changed into the dry roots of Morinda officinalis produced in Guangdong province and other places. The medicinal parts of Morindae Officinalis Radix in ancient and modern times are all roots， and the quality is better if it has many beads， thick flesh， and purple color. Ancient medical books recorded that it was usually harvested in February and August， dried in the shade， and used to remove the wood core. And the modern harvesting and processing method is to dig throughout the year， first remove the fibrous roots， dry in the sun until 60%-70% dry， gently beat flatten and dry in the sun. The processing methods of the past dynasties are mainly salt-， vinegar-， wine-processed， etc. Based on the systematic research of Morindae Officinalis Radix， from the perspective of clinical experience and safety and effectiveness， it is recommended that the famous classical formulas should be developed from the mainstream variety since modern times， namely Morindae Officinalis Radix.
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ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.
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ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.
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A method based on high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for the quantitative analysis of three active compounds and chemical fingerprint analyses of saccharides in Morindae officinalis radix. Ten batches of Morindae officinalis radix were collected from different plantations in the Guangdong region of China and used to establish the fingerprint. The samples were separated with a COSMOIL Sugar-D column (4.6 mm × 250 mm, 5 µm) by using gradient elution with water (A) and acetonitrile (B). In addition, Trapped-Ion-Mobility (tims) Time-Of-Flight (tims TOF) was used to identify saccharides of Morindae officinalis radix. Fingerprint chromatogram presented 26 common characteristic peaks in the roots of Morinda officinalis How, and the similarities were more than 0.926. In quantitative analysis, the three compounds showed good regression (r = 0.9995-0.9998) within the test ranges, and the recoveries of the method were in the range of 96.7-101.7%. The contents of sucrose, kestose and nystose in all samples were determined as 1.21-7.92%, 1.02-3.37%, and 2.38-6.55%, respectively. The developed HPLC fingerprint method is reliable and was validated for the quality control and identification of Morindae officinalis radix and can be successfully used to assess the quality of Morindae officinalis radix.
Subject(s)
Drugs, Chinese Herbal , Oligosaccharides , Scattering, Radiation , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Linear Models , Oligosaccharides/analysis , Oligosaccharides/isolation & purification , Principal Component Analysis , Reproducibility of ResultsABSTRACT
Morindae officinalis Radix (MOR) is a famous traditional Chinese medicine (TCM) and functional food material for invigorating kidneys and tonifying yang. Processed Morindae officinalis Radix (PMOR) is obtained by steaming MOR. Traditionally, the clinical effects are discrepant between processing and nonprocessing herbal medicines. MOR and PMOR are commonly used in both clinical practice and dietary supplements, and the effect of invigorating kidneys and tonifying yang of PMOR is stronger than MOR. To clarify the overall chemical composition and the difference of MOR and PMOR, a method was developed with an ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). Among the 110 identified components shared by MOR and PMOR, 55 compounds showed significant differences in contents. Among them, the contents of 29 components, including fructooligosaccharides, monotropein, deacetylasperulosidic acid, geniposide, and anthraquinone glycosides, were higher in MOR than in PMOR; the contents of 26 components, including difructose anhydride sucrose, and iridoid glycoside derivatives, were higher in PMOR than in MOR. Difructose anhydrides and iridoid glycoside derivatives were first discovered in PMOR. These results provided a scientific basis for research on the therapeutic material basis of MOR. It would provide a method for the comparison of processing and nonprocessing in Chinese medicines.
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Objective:To investigate the differences of main components of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt. Method:A total of 83 batches samples were collected in the market， including 41 batches of prepared Morindae Officinalis Radix，32 batches of Morindae Officinalis Radix processed with steaming and 10 batches of Morindae Officinalis Radix processed with salt. The contents of main components were determined with high performance liquid chromatography coupled with four-pole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS），and the differences were analyzed. Result:The main components of prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt were fructo-oligosaccharides （GF<italic>n</italic>），monotropein. The main components of Morindae Officinalis Radix processed with steaming were fructose，glucose，sucrose，and monotropein. The main differences of prepared Morindae Officinalis Radix and<italic> </italic>Morindae Officinalis Radix processed with steaming and salt were the contents of fructose，glucose，sucrose and GF2-GF11. The contents of GF2-GF11 in Morindae Officinalis Radix processed with steaming and salt were all lower than those in prepared Morindae Officinalis Radix，with extremely significant differences（<italic>P</italic><0.01）. The contents of fructose，glucose and sucrose in<italic> </italic>Morindae Officinalis Radix processed with steaming were significantly higher than those in prepared Morindae Officinalis Radix. The content of GF3 in each batch was higher than 40.0 mg·g<sup>-1</sup> in prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt，and significantly higher than the limit in<italic> Chinese Pharmacopoeia</italic>. However，there were only a few batches of Morindae Officinalis Radix processed with steaming in line with the requirements of <italic>Chinese Pharmacopoeia</italic>. The contents of monotropein in processing Morindae Officinalis Radix and Morindae Officinalis Radix processed with steaming and salt were 42.6，39.8，32.3 mg·g<sup>-1</sup>，respectively. The content of monotropein in prepared Morindae Officinalis Radix was higher than that in Morindae Officinalis Radix processed with steaming. The content of monotropein in Morindae Officinalis Radix processed with steaming was higher than that in Morindae Officinalis Radix<italic> </italic>processed with salt. Compared with the components of GF2-GF11，the effect of processing with steaming process and/or salt on monotropein content was relatively less. Conclusion:The contents of GF2-GF11 components in prepared Morindae Officinalis Radix were converted into fructose，glucose and sucrose after processing with steaming and/or salt. The results showed that the content limit of Morindae Officinalis Radix processed with steaming needs to be revised in line with the requirements of <italic>Chinese Pharmacopoeia </italic>for the quality control of Morindae Officinalis Radix. The results provide a reference basis for revising the quality standards and studying the pharmacodynamic material basis of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt.
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Morindae officinalis radix (MOR) is a famous Chinese herbal medicine which has long history of use in medicine and food. MOR and MOR with steaming process (PMOR) are the most commonly used forms in in clinical and health care. In order to establish a fast and mostly nondestructive quality control method for MOR, 183 beaches of MOR samples and 20 beaches of PMOR samples were collected commercially from major producing areas in Guangdong, Fujian and Guangxi Provinces of China. To predict main components of MOR, a calibration model was established based on near-infrared spectroscopy with partial least square regression. The model was optimized by compared the parameters of root mean square error of prediction (RMSEP), root mean square error of cross validation (RMSECV), coefficient of correlation (R2) and ratio of performance to deviation (RPD). Comparative studies were performed to evaluate the performance of models by different spectra preprocessing methods and different data set. The results showed that the model performance was improved with standard normal variate spectra preprocessing methods and when the data set contained both MOR and PMOR samples. A few PMOR samples were added to MOR samples data set the model predictive performance could be improved. The contents of 14 components were predicted in MOR with lower RMSEP and RMSECV, and higher R2 and RPD, including fructose (12.8 mg/g, 16.3 mg/g, 0.9873, 10.10), glucose (7.28 mg/g, 8.73 mg/g, 0.9611, 6.21 sucrose (9.24 mg/g, 9.10 mg/g, 0.8419, 1.75), GF2(9.42 mg/g, 11.3 mg/g, 0.8526, 2.03), GF3(7.98 mg/g, 9.20 mg/g, 0.8756, 2.74), GF4(6.81 mg/g, 8.93 mg/g, 0.8663, 3.06), GF5(8.13 mg/g, 8.85 mg/g, 0.9001, 3.06), GF6(6.40 mg/g, 6.95 mg/g, 0.9145, 3.27), GF7(5.53 mg/g, 6.15 mg/g, 0.9195, 3.57), GF8(5.40 mg/g, 6.02 mg/g, 0.9179, 3.31), GF9(3.00 mg/g,4.35 mg/g,0.9446, 5.03),GF10(4.08 mg/g, 5.34 mg/g, 0.8983, 3.62), GF11(8.97 mg/g, 7.70 mg/g, 0.8683, 2.01) and iridoid glycosides (4.12 mg/g, 5.51 mg/g, 0.8712, 2.43). The model established in this paper could predict 14 components of MOR. The results would provide a reference method for the quality control of Chinese medical materials and their process products.
Subject(s)
Fructose/analysis , Glucose/analysis , Iridoid Glycosides/analysis , Morinda/chemistry , Oligosaccharides/analysis , Spectroscopy, Near-Infrared , Sucrose/analysis , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Principal Component Analysis , Reproducibility of ResultsABSTRACT
Objective::To investigate the processing purpose of Morindae Officinalis Radix (MO), Euodiae Fructus (EF) and Polygalae Radix (PR) processed by Glycyrrhizae Radix et Rhizoma (Gly). Method::The content of dapsone in rat plasma was determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A)-water (B) for gradient elution (0-5 min, 10%-25%A; 5-20 min, 25%A) and detection wavelength was set at 292 nm. PK Solution 2.0 software was used to simulate pharmacokinetic parameters. Result::Within 300 min after dapsone was administrated, compared with the control (CTL) group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of MO (UMO) group. The elimination of dapsone was accelerated and its peak concentration (Cmax) was decreased in the processed products of MO with Gly (GMO) groups, and they had positive correlation with proportion of Gly in GMO. Compared with the CTL group, the elimination of dapsone was slowed down, and its plasma concentration was increased and its peak time (Tmax) was postponed in the unprocessed product of EF (UEF) group, while their Cmax and Tmax were changed in the processed products of EF with Gly (GEF) groups. Compared with the CTL group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of PR (UPR) group, while the elimination was accelerated and its plasma concentration was decreased in the processed products of PR with Gly (GPR) groups. Conclusion::The elimination of dapsone is slowed down in rats administered with UMO, UEF and UPR, while its elimination is accelerated in rats administered with the processed products of these three herbs with different proportions of Gly. Among the proportions, effect of processed products of these three herbs with 100∶6 (ratio of unprocessed product-Gly) on pharmacokinetics of dapsone is not significant.
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Objective: A method for identification of root cortex and woody core of Morindae Officinalis Radix (MOR) was established based on Ultra Performance Liquid Chromatography (UPLC) characteristic chromatogram and chemical pattern recognition technique. Methods: Using UPLC technique, the characteristic chromatograms of iridoids and oligosaccharides of root cortex and woody core of MOR were established, combined with the similarity analysis, variance analysis, cluster analysis, principal component analysis (PCA) methods for chemical pattern recognition research. Results: The UPLC characteristic chromatograms of iridoids and oligosaccharides of different parts of MOR were established, and 12 and 20 common characteristic peaks were confirmed, respectively. The UPLC characteristic chromatograms of root cortex and woody core of MOR were obviously different. Conclusion: UPLC characteristic chromatograms of iridoids and oligosaccharides of MOR combined with chemical pattern recognition analysis method can reflect the difference of root cortex and woody core of MOR integrally, comprehensively and truly, which provides more sufficient basis for the necessity of removing woody core from MOR.
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Objective: To study the effect of Morindae Officinalis Radixe extract from different origins on the vascular growth of damaged internodes of zebra fish, so as to discuss regional differences of Morindae Officinalis Radix from different origins. Method: A zebra fish model of internode vascular injury induced by PTK787 was established to study the effect of Morindae Officinalis Radix extract from Guangdong, Guangxi and Fujian on the growth of internode vascular in zebra fish, and detect the number of internode angiogenesis in zebra fish treated with PTK787.On the basis of principal component analysis, relative angiogenesis rate was taken as the index, and the experimental data was analyzed by SPSS 17.0 statistical software. Result: The extract of Morindae Officinalis Radix had a repairing effect on the growth of damaged internode blood vessels of zebra fish, and the extracts of Morindae Officinalis Radix from Guangdong Gaoyao, and Guangdong Yunan, were better than those from other producing areas. The average number of internode angiogenesis was 9, the average length of angiogenesis was 448-504 μm, and the relative rate of angiogenesis was between 7.9% and 8.86%. The results of cluster analysis showed that Morindae Officinalis Radix of 12 different origins were classified into four categories, Guangdong Deqing, and Guangdong Gaoyao, Guangxi Cangwu and Guangxi Cenxi, and Guangdong Yunan, and Guangxi Cangwu could be better clustered. Conclusion: There are obvious regional differences in the pro-angiogenic effect of Morindae Officinalis Radix from different origins. Relevant studies provide basic data for the quality assessment of Morindae Officinalis Radix medicinal materials.
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Objective: To screen the extraction process of Wuzi Yanzong plus and minus Recipe (WYR) by pharmacodynamics and long-term toxicity test, and optimize it by orthogonal experiment to determine the best extraction process of WYR. Methods Long-term toxicity experiments were carried out on different processed WYR samples. The penile erection latency and sex hormone levels in rats with kidney-yang deficiency were used as pharmacodynamic indicators. The results of long-term toxicity and efficacy experiments were combined to screen out the optimal process plan. Using the content of ingredients and the rate of ointment as indicators, the orthogonal test was used to screen the optimal level of different extraction processes and verified. Results Long-term toxicity test results showed that all mice survived healthily and no obvious toxicity was observed. The results of pharmacodynamic experiments showed that the extracts of Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were refluxed with 70% ethanol, and the remaining medicinal materials were extracted with 70% ethanol as the solvent to obtain the best extraction method of WYR. The suitable extraction process was as follows: Morindae Officinalis Radix, Eucommiae Cortex, Taxilli Herba, Dipsaci Radix, and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle were extracted three times with 10 times 70% ethanol for 2.0 h each time; The remaining medicinal materials were percolated with 10 times 70% ethanol at 1.5 mL/min. The medicinal herbs were firstly soaked for 16 h before percolation. Conclusion WYR can significantly improve the penile erection latency and sex hormone levels in rats with kidney yang deficiency. The optimal process conditions are reasonable and feasible, which provides a basis for the follow-up development of WYR.
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Objective: To investigate the stability of icariin, 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside in Huangmai mixture.Methods: TLC was used to identify astrogaloside IV and Morindae officinalis Radix, and HPLC was used to determine the contents of icariin, 2,3,5,4'-retrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside.The stability of Huangmai mixture under the conditions of accelerated testing and long-term testing was assessed.Results: Astrogaloside IV, Morindae officinalis Radix and icariin showed good stability during the accelerated testing and long-term testing.The contents of 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside declined quickly during the stability testing.Conclusion: Hyperoside and 2,3,5,4'-retrahydroxy-stilbene-2-O-β-D-glucoside are unstable in Huangmai mixture.A novel solid dosage form of Huangmai should be developed to obtain better stability.
