In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after co...In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.展开更多
Doxorubicin(DOX)is a widely employed tumor therapy,yet its substantial toxic side effects pose a considerable challenge.Bletilla striata has demonstrated efficacy in preventing and treating these toxic side effects in...Doxorubicin(DOX)is a widely employed tumor therapy,yet its substantial toxic side effects pose a considerable challenge.Bletilla striata has demonstrated efficacy in preventing and treating these toxic side effects in clinical practice,with polysaccharides identified as the principal active component.In the present study,16 fractions of B.striata polysaccharides(BsPs)were extracted using diverse methods,including hot-water extraction(HWE),ultrasonic-assisted extraction(UAE),enzyme-assisted extraction(EAE),dilute acid-water extraction(ACWE),and dilute alkali-water extraction(ALWE).These extractions were subsequently precipitated at final ethanol concentrations of 80%and 95%,respectively.The investigation encompassed yields,total carbohydrate content(TCC),total protein content(TPC),preliminary structural properties,and anti-DOX myocardial cytotoxic activity.Results indicated that the extraction method significantly influenced the physicochemical properties,associated functional properties,and anti-DOX myocardial cytotoxic activity of BsPs.HWE and UAE yielded higher BsPs quantities.The relative molecular weight(RMW)distribution of BsPs differed notably between HWE or UAE,EAE,ACWE,and ALWE.The RMW of primary BsPs obtained from HWE and UAE(1.9×10^(7)-1.7×10^(7) Da)exceeded that from EAE(7.5×10^(3)-2.8×10^(4) Da)and ALWE(5.1×10^(4)-1.7×10^(4) Da),with smaller molecular weights primarily precipitated by higher ethanol concentrations.BsPs were composed of Man and Glu,with partial fractions containing small amounts of Gal or Ara,displaying varying molar ratios.Notably,BsPs from ACWE exhibited the most significant structural differences,lacking 1,4-α-D-Glcp and a triple-helical structure.Furthermore,BsPs obtained from HWE,UAE,and EAE demonstrated heightened anti-DOX myocardial cytotoxic activity compared to other methods.This study underscored the influence of extraction methods on BsPs’structure and myocardial protective activity,offering a foundation for exploring structural diversity and employing specific extraction methods to extract polysaccharides with robust myocardial protective properties efficiently.展开更多
Objective: To study the protective activities of the dietary malate esters derivatives of Bletilla striata against SiO_(2)nanoparticles-induced A549 cell lines and its mechanism action.Methods: The components were iso...Objective: To study the protective activities of the dietary malate esters derivatives of Bletilla striata against SiO_(2)nanoparticles-induced A549 cell lines and its mechanism action.Methods: The components were isolated and elucidated by spectroscopic methods such as 1D NMR and 2D NMR. And MTT assays was used to tested these components on the A549 cell survival rates and ROS or proteins levels were detected by Western blotting.Results: A new glucosyloxybenzyl 2-isobutylmalate(a malate ester derivative), along with 31 known compounds were isolated and identified from n-BuOH extract of EtOH extract of B. striata. Among them,compounds 3, 4, 11, 12 and 13 possessed noteworthy proliferative effects for damaged cells, with ED50of 14.0, 13.1, 3.7, 11.6 and 11.5 μmol/L, respectively, compared to positive control resveratrol(ED50, 14.7 μmol/L). Militarine(8) prominently inhibited the intracellular ROS level, and increased the expression of Nrf2 and its downstream genes(HO-1 and γ-GCSc). Furthermore, Nrf2 activation mediates the interventional effects of compound 8 against SiO_(2)nanoparticles(nm SiO_(2))-induced lung injury.Moreover, treatment with compound 8 significantly reduced lung inflammation and oxidative stress in nm SiO_(2)-instilled mice. Molecular docking experiment suggested that 8 bound stably to the HO-1 protein by hydrogen bond interactions.Conclusion: The dietary malate esters derivatives of B. striata could significantly increase the viability of nm SiO_(2)-induced A549 cells and decrease the finer particles-induced cell damages. Militarine is especially promising compound for chemoprevention of lung cancer induced by nm SiO_(2)through activation of Nrf2 pathway.展开更多
Rhizoma Bletillae is in high demand as a traditional Chinese medicine, and the natural resources have been severely damaged due to excessive exploitation. Because Bletilla striata seeds are small and have no endosperm...Rhizoma Bletillae is in high demand as a traditional Chinese medicine, and the natural resources have been severely damaged due to excessive exploitation. Because Bletilla striata seeds are small and have no endosperm, the seed germination rate is low in natural conditions. Traditional division propagation could not meet the demands of large-scale cultivation. Tissue culture can provide many seedlings in a short time and is more effective and convenient than other methods. Most studies on tissue culture of B. striata selected seeds as explants. This review summarized the processes of the aseptic seed germination pathway. It included such stages as seed germination, proliferation of clusters of buds, induction of root- ing and transplanting of seedlings. Influential factors as well as optimum combination and concentration of the plant growth regulators of each stage were also summarized. The further research on tissue culture of B. striata was also prospected.展开更多
基金supported by the National Basic Research Program of China("973Program)(No2011CB505304)the Youth Scientific Research Project of Anhui Academy of Medical Science(YKY2018003)
文摘In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.
基金Science and Technology Foundation of Guizhou Province(Grant No.QKHJC-ZK[2023]YB524)Science and Technology Plan Project of Guizhou,China(Grant No.QKPTRC[2019]035)+2 种基金Science and Technology Foundation of Guizhou Province(Grant No.QKH[2019]1346)Science and Technology Department of Zunyi city of Guizhou province of China(Grant No.[2020]7)Undergraduate Training Program for Innovation and Entrepreneurship of Zunyi Medical University(Grant No.S202310661248).
文摘Doxorubicin(DOX)is a widely employed tumor therapy,yet its substantial toxic side effects pose a considerable challenge.Bletilla striata has demonstrated efficacy in preventing and treating these toxic side effects in clinical practice,with polysaccharides identified as the principal active component.In the present study,16 fractions of B.striata polysaccharides(BsPs)were extracted using diverse methods,including hot-water extraction(HWE),ultrasonic-assisted extraction(UAE),enzyme-assisted extraction(EAE),dilute acid-water extraction(ACWE),and dilute alkali-water extraction(ALWE).These extractions were subsequently precipitated at final ethanol concentrations of 80%and 95%,respectively.The investigation encompassed yields,total carbohydrate content(TCC),total protein content(TPC),preliminary structural properties,and anti-DOX myocardial cytotoxic activity.Results indicated that the extraction method significantly influenced the physicochemical properties,associated functional properties,and anti-DOX myocardial cytotoxic activity of BsPs.HWE and UAE yielded higher BsPs quantities.The relative molecular weight(RMW)distribution of BsPs differed notably between HWE or UAE,EAE,ACWE,and ALWE.The RMW of primary BsPs obtained from HWE and UAE(1.9×10^(7)-1.7×10^(7) Da)exceeded that from EAE(7.5×10^(3)-2.8×10^(4) Da)and ALWE(5.1×10^(4)-1.7×10^(4) Da),with smaller molecular weights primarily precipitated by higher ethanol concentrations.BsPs were composed of Man and Glu,with partial fractions containing small amounts of Gal or Ara,displaying varying molar ratios.Notably,BsPs from ACWE exhibited the most significant structural differences,lacking 1,4-α-D-Glcp and a triple-helical structure.Furthermore,BsPs obtained from HWE,UAE,and EAE demonstrated heightened anti-DOX myocardial cytotoxic activity compared to other methods.This study underscored the influence of extraction methods on BsPs’structure and myocardial protective activity,offering a foundation for exploring structural diversity and employing specific extraction methods to extract polysaccharides with robust myocardial protective properties efficiently.
基金financially supported by National Natural Science Foundation of China,China(Grant No.81872768,81673323,82003630)Scientific Research Fund of Liaoning Province Education Department,Liaoning(LJKZ0949)+1 种基金Special Fund of Research Institute of Drug Regulatory Science Research Shenyang Pharmaceutical University,Liaoning(2021jgkx010)State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources(Guangxi Normal University),Guangxi(CMEMR2022-B04)。
文摘Objective: To study the protective activities of the dietary malate esters derivatives of Bletilla striata against SiO_(2)nanoparticles-induced A549 cell lines and its mechanism action.Methods: The components were isolated and elucidated by spectroscopic methods such as 1D NMR and 2D NMR. And MTT assays was used to tested these components on the A549 cell survival rates and ROS or proteins levels were detected by Western blotting.Results: A new glucosyloxybenzyl 2-isobutylmalate(a malate ester derivative), along with 31 known compounds were isolated and identified from n-BuOH extract of EtOH extract of B. striata. Among them,compounds 3, 4, 11, 12 and 13 possessed noteworthy proliferative effects for damaged cells, with ED50of 14.0, 13.1, 3.7, 11.6 and 11.5 μmol/L, respectively, compared to positive control resveratrol(ED50, 14.7 μmol/L). Militarine(8) prominently inhibited the intracellular ROS level, and increased the expression of Nrf2 and its downstream genes(HO-1 and γ-GCSc). Furthermore, Nrf2 activation mediates the interventional effects of compound 8 against SiO_(2)nanoparticles(nm SiO_(2))-induced lung injury.Moreover, treatment with compound 8 significantly reduced lung inflammation and oxidative stress in nm SiO_(2)-instilled mice. Molecular docking experiment suggested that 8 bound stably to the HO-1 protein by hydrogen bond interactions.Conclusion: The dietary malate esters derivatives of B. striata could significantly increase the viability of nm SiO_(2)-induced A549 cells and decrease the finer particles-induced cell damages. Militarine is especially promising compound for chemoprevention of lung cancer induced by nm SiO_(2)through activation of Nrf2 pathway.
基金supported by grants from the National Natural Science Foundation of China (No.81673552)CAMS Innovation Fund for Medical Sciences (CIFMS,2016-I2M-3-016)
文摘Rhizoma Bletillae is in high demand as a traditional Chinese medicine, and the natural resources have been severely damaged due to excessive exploitation. Because Bletilla striata seeds are small and have no endosperm, the seed germination rate is low in natural conditions. Traditional division propagation could not meet the demands of large-scale cultivation. Tissue culture can provide many seedlings in a short time and is more effective and convenient than other methods. Most studies on tissue culture of B. striata selected seeds as explants. This review summarized the processes of the aseptic seed germination pathway. It included such stages as seed germination, proliferation of clusters of buds, induction of root- ing and transplanting of seedlings. Influential factors as well as optimum combination and concentration of the plant growth regulators of each stage were also summarized. The further research on tissue culture of B. striata was also prospected.
