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Molecular characterization and structure basis of a malonyltransferase with both substrate promiscuity and catalytic regiospecificity from Cistanche tubulosa

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Acta Pharmaceutica Sinica B 2024年第5期14卷 2333-2348页

作者： Xiao Liu Yuyu Liu Xiping Xu Wenqian Huang Yaru Yan Yingxia Wang Weisheng Tian Ting Mo Xiaoxue Cui Jun Li She-Po Shi Pengfei Tu Modern research Center for Traditional chinese medicine Beijing Research Institute of Chinese MedicineBeijing University of Chinese MedicineBeijing 100029China Modern research Center for Traditional chinese medicine School of Chinese Materia MedicaBeijing University of Chinese MedicineBeijing 100029China

Enzymatic malonylation of natural glycosides provides a promising alternative method for drug-like malonylated glycosides ***,the catalytic potential and structural basis of plant malonyltransferase are far from being fully *** work identified a new malonyltransferase CtMaT1 from Cistanche *** displayed unprecedented mono-and/or di-malonylation activity toward diverse glucosides with different aglycons.A“one-pot”system by CtMaT1 and a malonyl-CoA synthetase was established to biosynthesize nine new malonylated *** investigations revealed that CtMaT1 possesses an adequately spacious acyl-acceptor pocket capable of accommodating diverse ***,it recognizes malonyl-CoA through strong electrotactic and hydrogen ***/MM calculation revealed the H167-mediated SN2 reaction mechanism of CtMaT1,while dynamic simulations detected the formation of stable hydrogen bonds between the glucose-6-OH group and H167,resulting in its high malonylation *** energy profiles of two isomeric glycosides highlighted lower reaction energy barriers towards glucoside substrates,emphasizing CtMaT1's preference for ***,a mutant CtMaT1H36A with notably increased di-malonylation activity was *** underlying molecular mechanism was illuminated through MM/GBSA binding free energy *** study significantly advances the understanding of plant acyltransferases from both functional and protein structural perspectives,while also providing a versatile tool for enzymatic malonylation applications in pharmacology.

关键词： Plant acyltransferase Malonyltransferase Malonylated glycosides Enzymatic catalysis Enzyme promiscuity

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Shexiang Tongxin Dropping Pill Promotes Angiogenesis through VEGF/eNOS Signaling Pathway on Diabetic Coronary Microcirculation Dysfunction

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chinese Journal of Integrative medicine 2024年第10期30卷 886-895页

作者： CUI Xin-yu LIU Tian-hua BAI Ya-li ZHANG Meng-di LI Guo-dong ZHANG Yu-ting YUAN Yue-ying ZHANG Ya-wen YU Li-shuang HAN Li-na WU Yan School of Traditional chinese medicine Beijing University of Chinese Medicine Key Laboratory of Traditional chinese medicine Syndrome and Formula Ministry of Education Beijing University of Chinese Medicine Beijing research Institute of chinese medicine Beijing University of Chinese Medicine

Objective: To study the effect of Shexiang Tongxin Dropping Pill(STDP) on angiogenesis in diabetic cardiomyopathy mice with coronary microcirculation dysfunction(CMD). Methods: According to a random number table, 6 of 36 SPF male C57BL/6 mice were randomly selected as the control group, and the remaining 30 mice were injected with streptozotocin intraperitoneally to replicate the type 1 diabetes model. Mice successfully copied the diabetes model were randomly divided into the model group, STDP low-dose group [15 mg/(kg·d)], medium-dose group [30 mg/(kg·d)], high-dose group [60 mg/(kg·d)], and nicorandil group [15 mg/(kg·d)], 6 in each group. The drug was given by continuous gavage for 12 weeks. The cardiac function of mice in each group was detected at the end of the experiment, and coronary flow reserve(CFR) was detected by chest Doppler technique. Pathological changes of myocardium were observed by hematoxylin-eosin staining, collagen fiber deposition was detected by masson staining, the number of myocardial capillaries was detected by platelet endothelial cell adhesion molecule-1 staining, and the degree of myocardial hypertrophy was detected by wheat germ agglutinin staining. The expression of the vascular endothlial growth factor(VEGF)/endothelial nitric oxide synthase(eNOS) signaling pathway-related proteins in myocardial tissue was detected by Western blot. Results: Compared with the model group, medium-and high-dose STDP significantly increased the left ventricular ejection fraction and left ventricular fraction shortening(P<0.01), obviously repaired the disordered cardiac muscle structure, reduced myocardial fibrosis, reduced myocardial cell area, increased capillary density, and increased CFR level(all P<0.01). Western blot showed that high-dose STDP could significantly increase the expression of VEGF and promote the phosphorylation of vascular endothelial growth factor receptor 2, phosphoinositide 3-kinase, protein kinase B, and eNOS(P<0.05 or P<0.01). Co

关键词： Shexiang Tongxin Dropping Pill angiogenesis coronary microcirculation dysfunction diabetic cardiomyopathy type 1 diabetes

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Brazilin Actuates Ferroptosis in Breast Cancer Cells via p53/SLC7A11/GPX4 Signaling Pathway

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chinese Journal of Integrative medicine 2024年第11期30卷 1001-1006页

作者： HE Dan TAN Xiao-ning LI Lin-pei GAO Wen-hui TIAN Xue-fei ZENG Pu-hua Hunan Academy of chinese medicine School of Integrated chinese and Western medicine Hunan University of Chinese Medicine Scientific research Department The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine Oncology Department The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine School of Traditional chinese medicine Hunan University of Chinese Medicine

Objective:To investigate the mechanism of induction of ferroptosis by brazilin in breast cancer ***:Breast cancer 4T1 cells were divided into 6 groups:control,brazilin 1/2 half maximal inhibitory concentration(IC50),IC50,2×IC50,erastin（10 μg/mL） and capecitabine（10 μg/mL） *** effect of brazilin on the proliferation of 4T1 cells was detected by cell counting kit-8 assay,and the treatment dose of brazilin was *** effect of brazilin on the mitochondrial morphology of 4T1 cells,and the mitochondrial damage was evaluated under electron *** levels of Fe2+,reactive oxygen species（ROS）,malondialdehyde（MDA）,glutathione（GSH） and glutathione peroxidase 4（GPX4） were estimated using various detection *** invasion and migration abilities of 4T1 cells were detected by scratch assay and transwell *** expressions levels of tumor protein p53, solute carrier family 7 member 11（SLC7A11）,GPX4 and acyl-CoA synthetase long-chain family member 4（ACSL4） proteins were quantified by Western blot ***:Compared to the control group,the 10(1/2 IC50),20(IC50) and 40(2×IC50) μg/mL brazilin,erastin,and capecitabine groups showed a significant decrease in the cell survival rate,invasion and migration abilities,GSH,SLC7A11 and GPX4 protein expression levels,and mitochondrial volume and ridge（P<0.05）,and a significant increase in the mitochondria membrane density,Fe2+,ROS and MDA levels,and p53 and ACSL4protein expression levels（P<0.05）.Conclusions:Brazilin actuated ferroptosis in breast cancer cells,and the underlying mechanism is mainly associated with the p53/SLC7A11/GPX4 signaling pathway.

关键词： chinese medicine brazilin breast cancer ferroptosis p53/SLC7A11/GPX4 axis

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Zinc-based metal-organic frameworks as efficient carriers for anticancer drug to reduce toxicity and increase efficacy

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Rare Metals 2024年第10期43卷 5152-5163页

作者： Dong-Wei Ma Jing-Sheng Lu Xiang-Xin Cao Yan-Wei Cheng Gang Wang Zi-Qian Zhang Bo-Cheng Chen Ning Lin Qing Chen College of Pharmacy Guangxi Scientific Research Center of Traditional Chinese MedicineGuangxi University of Chinese Medicine Guangxi Zhuang Yao medicine Center of Engineering and Technology

The rational design of a specific co-drug delivery platform that can address the unavoidable resistance, toxic side effects and low targeting efficiency of traditional cancer treatments is of great ***, Zn-based MOF-zeolitic imidazole framework-90(ZIF-90) was selected as the drug delivery carrier, with the cancer therapeutic drug mercaptopurine(6-MP) and glucose oxidase(GOD) as the drug models, hyaluronic acid(HA) was used for protection and targeted delivery, which designed and fabricated an intelligent drug delivery platform(6-MP@ZIF-90@GOD@HA). This platform aims to reduce the toxic side effects of 6-MP and enhance its efficacy while improving the targeting ability of GOD,thereby further enhancing the treatment outcome. Notably,results from both in vitro and in vivo experiments demonstrate that the targeted synergistic chemo/reactive oxygen species(ROS)-mediated/starvation therapy inhibited the cancer cell growth while reducing the chemotherapy toxicity, which provides new possibilities for the development of more precise and effective treatment strategies.

关键词： Cancer treatment Combination therapy Drug delivery Metal-organic framework Reduced toxicity

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Application of nanoscale metal–organic frameworks in tumor immunotherapy

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Rare Metals 2024年第10期43卷 4867-4883页

作者： Xiao-Xuan Zeng Jing-Sheng Lu Dong-Wei Ma Yu-Ting Huang Ling Chen Gang Wang Qing Chen Ning Lin College of Pharmacy Guangxi Scientific Research Center of Traditional Chinese MedicineGuangxi University of Chinese Medicine Guangxi Zhuang Yao medicine Center of Engineering and Technology

Immunotherapy has attracted wide attention because it can not only kill tumor cells but also effectively prevent tumor recurrence. However, due to the poor efficacy of the monotherapy in tumor immunotherapy, combination therapies are needed to enhance their efficacy,such as chemodynamic therapy/immunotherapy, photodynamic therapy/immunotherapy, sonodynamic therapy/immunotherapy and so on. Nanoscale metal–organic frameworks(nMOFs) are widely used for drug delivery because of their rich and variable composition, high drug loading capacity and good biodegradability. Compared with other conventional drug delivery systems(DDSs),nMOFs play more advantageous role in inducing immune cell death of tumor cells and combination therapies. This article focuses on the application of n MOFs in tumor immunotherapy in terms of their use as gatekeepers, supplementary agents and drug carriers, respectively, with the aim of providing some ideas and references for the design of smart nanoscale DDS for tumor immunotherapy.

关键词： Drug delivery system Immunotherapy Metal–organic frameworks Tumor

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Triple three-dimensional MS/MS spectrum facilitates quantitative ginsenosides-targeted sub-metabolome characterization in notoginseng

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Acta Pharmaceutica Sinica B 2024年第9期14卷 4045-4058页

作者： Ke Zhang Jinru Jia Ting Li Wenjing Liu Pengfei Tu Jian-Bo Wan Jun Li Yuelin Song Modern research Center for Traditional chinese medicine Beijing Research Institute of Chinese MedicineBeijing University of Chinese MedicineBeijing 100029China SCIEX China Beijing 100015China School of Pharmacy Henan University of Chinese MedicineZhengzhou 450046China State Key Laboratory of Quality research in chinese medicine Institute of Chinese Medical SciencesUniversity of MacaoTaip1Macao 999078China

Although serving as the workhorse,MS/MS cannot fully satisfy the analytical requirements of quantitative sub-metabolome *** more information intrinsically correlates to more structural and concentration clues,here,efforts were devoted to comprehensively tracing and deciphering MS/MS behaviors through constructing triple three-dimensional(33D)-MS/MS ***-targeted metabolomics of notoginseng,one of the most famous edible medicinal plants,was employed as a *** authentic ginsenosides were deployed to build the correlations between 33D-MS/MS spectra and structure/concentration *** assaying ginsenosides with progressive concentrations using QTOF-MS to configure 1st 3D spectrum,the generations of MS1 spectral signals,particularly multi-charged multimer anions,e.g.,[2Me2H]2e and[2Mt2HCOO]2e ions,relied on both concentration and the amount of sugar *** programming progressive collision energies to the front collision cell of Qtrap-MS device to gain 2nd 3D spectrum,optimal collision energy(OCE)corresponding to the glycosidic bond fission was primarily correlated with the masses of precursor and fragment ions and partially governed by the glycosidation *** quantitative relationships between OCEs and masses of precursor and fragment ions were utilized to build large-scale quantitative program for *** applying progressive exciting energies to the back collision chamber to build 3rd 3D spectrum,the fragment ion and the decomposition product anion exhibited identical dissociation trajectories when they shared the same molecular *** ginsenosides-focused quantitative metabolomics,significant differences occurred for sub-metabolome amongst different parts of *** differential ginsenosides were confirmatively identified by applying the correlations between 33D-MS/MS spectra and ***,33D-MS/MS spectrum covers all MS/MS behaviors and dramatically faci

关键词： Quantitative submetabolome characterization Ginsenosides Structural identification Notoginseng Full exciting energy ramp-MS3 spectrum

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Protective Effect of Silibinin on Lipopolysaccharide-Induced Endotoxemia by Inhibiting Caspase-11-Dependent Cell Pyroptosis

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chinese Journal of Integrative medicine 2024年第10期30卷 917-926页

作者： OU Jin-ying LIU Shan-hong TANG Dong-kai SHI Ling-zhu YAN Li-jun HUANG Jing-yan ZOU Li-fang QUAN Jing-yu YOU Yan-ting CHEN Yu-yao YU Lin-zhong LU Zi-bin Third Level research Laboratory of State Administration of Traditional chinese medicine School of Traditional Chinese Medicine Southern Medical University Guangdong Provincial Key Laboratory of chinese medicine Pharmaceutics School of Traditional chinese medicine Southern Medical University

Objective: To explore the protective effect and the underlying mechanism of silibinin(SIB), one of the active compounds from Silybum marianum(L.) Gaertn in endotoxemia. Methods: Mouse peritoneal macrophage were isolated via intraperitoneally injection of BALB/c mice with thioglycolate medium. Cell viability was assessed using the cell counting kit-8, while cytotoxicity was determined through lactate dehydrogenase cytotoxicity assay. The protein expressions of interleukin(IL)-1α, IL-1β, and IL-18 were determined by enzyme-linked immunosorbent assay. Intracellular lipopolysaccharide(LPS) levels were measured by employing both the limulus amoebocyte lysate assay and flow cytometry. Additionally, proximity ligation assay was employed for the LPS and caspase-11 interaction. Mice were divided into 4 groups: the control, LPS, high-dose-SIB(100 mg/kg), and low-dose-SIB(100 mg/kg) groups(n=8). Zebrafish were divided into 4 groups: the control, LPS, high-dose-SIB(200 μmol/L), and low-dose-SIB(100 μmol/L) groups(n=30 for survival experiment and n=10 for gene expression analysis). The expression of caspase-11, gasdermin D(GSDMD), and N-GSDMD was determined by Western blot and the expressions of caspy2, gsdmeb, and IL-1β were detected using quantitative real-time PCR. Histopathological observation was performed through hematoxylineosin staining, and protein levels in bronchoalveolar lavage fluid were quantified using the bicinchoninicacid protein assay. Results: SIB noticeably decreased caspase-11 and GSDMD-mediated pyroptosis and suppressed the secretion of IL-1α, IL-1β, and IL-18 induced by LPS(P<0.05). Moreover, SIB inhibited the translocation of LPS into the cytoplasm and the binding of caspase-11 and intracellular LPS(P<0.05). SIB also attenuated the expression of caspase-11 and N-terminal fragments of GSDMD, inhibited the relative cytokines, prolonged the survival time, and up-regulated the survival rate in the endotoxemia models(P<0.05). Conclusions: SIB can inhibit pyrop

关键词： caspase-11 endotoxemia inflammation pyroptosis silibinin

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Rapid Simultaneous Determination of Four Ganoderic Acids in Ganoderma(chinese Name:Lingzhi)by Direct Infusion-Multiple Reaction Monitoring Cubed

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Journal of Analysis and Testing 2024年第1期8卷 52-62页

作者： Nian Wu Bo Peng Ting Li Pengfei Tu Shengpeng Wang Bo Li Wenjing Liu Yuelin Song Modern research Center for Traditional chinese medicine Beijing Research Institute of Chinese MedicineBeijing University of Chinese MedicineBeijing 100029China Amway(China)Botanical research Center Wuxi 214011China State Key Laboratory of Quality research in chinese medicine Institute of Chinese Medical SciencesUniversity of MacaoTaipa 999078China Amway(Shanghai)Innovation&Science Co. LtdShanghai 201508China School of Pharmacy Henan University of Chinese MedicineZhengzhou 450046China

Attributing to the rapid demand expansion for the edible medicinal materials in the market,the limited throughput of highperformance liquid chromatography-multiple reaction monitoring(HPLC-MRM)cannot fully address the measurement workload for a huge number of testing ***,it is urgent to pursue more efficient approaches for the quality *** of the greater selectivity of MRM cubed(MRM^(3))over MRM,there might be a chance to omit the time-intensive LC *** current study,we attempted to develop a direct infusion(DI)-MRM^(3) program,and the applicability was thereafter assessed through simultaneous determination of four ganoderic acids(GAs)in one of the most famous tonic herbal medicines namely Ganoderma(chinese name:Lingzhi).Primary parameters such as Q1>Q3>QLIT ion transitions,collision energy(CE),and excitation energy were optimized by programming online energy-resolved mass spectrometry with authentic compounds.A single DI-MRM measurement merely costed four minutes,and in spite of the wide occurrences of isomers,satisfactory selectivity was *** validation assays demonstrated the method to be sensitive,precise,accurate,and *** quantitative results from DI-MRM^(3) were also justified by conducting LC-MRM measurements in *** differences occurred for the content patterns between the two original sources namely Ganoderma lucidum and ***,and,moreover,either cultivar or harvest time showed dramatical influence on the quantitative features of the four targeted *** importantly,DI-MRM3 is a meaningful analytical option for rapid quantitative analysis of herbal medicines,because of the comparable reliability,nonetheless,less consumptions of both measurement time and solvent,compared with LC-MRM.

关键词： Direct infusion Multiple reaction monitoring cubed High-throughput quantitative analysis Ganoderic acids Ganoderma

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Fu-Zheng-Yi-Liu Formula inhibits the stem cells and metastasis of prostate cancer via tumor-associated macrophages/C-C motif chemokine ligand 5 pathway in tumor microenvironment

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chinese Journal of Natural medicines 2024年第6期22卷 501-514页

作者： CHEN Chiwei HUANG Renlun WANG Neng ZHENG Yifeng ZHOU Jianfu YANG Bowen WANG Xuan ZHANG Juping PAN Bo CHEN Zhiqiang WANG Shengqi WANG Zhiyu XIANG Songtao The research Center of Integrative Cancer medicine Discipline of Integrated Chinese and Western Medicinethe Second Clinical College of Guangzhou University of Chinese MedicineGuangzhou 510000China The First Clinical College of Guangzhou University of chinese medicine Guangzhou 510000China Guangdong Provincial Key Laboratory of Clinical research on Traditional chinese medicine Syndrome Guangdong Provincial Academy of Chinese Medical SciencesGuangdong Provincial Hospital of Chinese MedicineGuangzhou 510000China Guangdong-Hong Kong-Macao Joint Lab on chinese medicine and Immune Disease research Guangzhou University of Chinese MedicineGuangzhou 510000China The research Center for Integrative medicine School of Basic Medical SciencesGuangzhou University of Chinese MedicineGuangzhou 510000China State Key Laboratory of Dampness Syndrome of chinese medicine The Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou 510000China

Prostate cancer(PCa)is the second most common malignancy among men *** Fu-Zheng-Yi-Liu(FZYL)Formula has been widely utilized in the treatment of *** study investigates whether the FZYL Formula can inhibit PCa by tar-geting the TAMs/CCL5 *** conducted in vitro co-cultures and in vivo co-injections of PCa cells and TAMs to mimic their *** showed that the FZYL Formula significantly reduced the proliferation,colony formation,subpopulations of PCSCs,and sphere-formation efficacy of PCa cells,even in the presence of TAM ***,the Formula markedly decreased the migration,invasion,and epithelial-mesenchymal transition(EMT)of PCa cells induced by *** FZYL Formula also reversed M2 phenotype polarization in TAMs and dose-dependently reduced their CCL5 expression and secretion,with minimal cytotoxicity *** studies confirmed that the TAMs/CCL5 axis is a critical target of the FZYL Formula,as the addition of exogen-ous CCL5 partially reversed the formula’s inhibitory effects on PCSCs self-renewal in the co-culture ***,the Formula also significantly inhibited the growth of PCa xenografts,bone metastasis,and PCSCs activity in vivo by targeting the TAMs/CCL5 ***,this study not only elucidates the immunomodulatory mechanism of the FZYL Formula in PCa therapy but also highlights the TAMs/CCL5 axis as a promising therapeutic target.

关键词： Traditional chinese medicine Tumor-associated macrophages Prostate cancer stem cells CCL5 Tumor microenvironment

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Electroacupuncture ameliorates blood-brain barrier disruption after ischemic stroke through histone acetylation regulation at the matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 2 genes

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Journal of Traditional chinese medicine 2024年第4期44卷 734-744页

作者： CHEN Yonglin OUYANG Ling MENG Lingling WU Bufan PENG Rou LIU Sitong HOU Dan WANG Yaling JING Xinyue LU Shengfeng FU Shuping Key Laboratory of Acupuncture and medicine research of Ministry of Education Nanjing University of Chinese MedicineNanjing 210023China

OBJECTIVE:To explore whether the regulation of matrix metalloproteinase 9(MMP-9)/tissue inhibitors of MMPs(TIMPs)gene expression through histone acetylation is a possible mechanism by which electroacupuncture(EA)protects blood-brain barrier(BBB)integrity in a middle cerebral artery occlusion(MCAO)rat ***:Male Sprague-Dawley rats were divided into four groups:the sham group,the MCAO group,the MCAO+EA(MEA)group,and the MCAO+EA+HAT inhibitor(HATi)*** MCAO model was generated by blocking the middle cerebral *** was applied to Baihui(GV20).Samples were collected 1 or 3 d after *** function scores and Evans blue extravasation were employed to evaluate the poststroke *** effect of EA on MMP-9/TIMPs gene expression was assessed by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and chromatin immunoprecipitation(ChIP).RESULTS:Our results showed that EA treatment prominently improved neurological function and ameliorated BBB *** RT-qPCR assay showed that EA reduced the expression of MMP-9 and promoted TIMP-2 mRNA expression,but HATi reversed these effects of *** addition,ChIP results revealed that EA decreased the enrichment of H3K9ace/H3K27ace at MMP-9 promoters and notably stimulated the recruitment of H3K9ace/H3K27ace at TIMP-2 ***:EA treatment at Baihui(GV20)regulates the transcription of MMP-9 and TIMP-2 through histone acetylation modification in the acute stage of stroke,which preserves the structural integrity of the BBB in MCAO *** findings suggested that the histone acetylation-mediated transcriptional activity of target genes may be a crucial mechanism of EA treatment in stroke.

关键词： blood-brain barrier matrix metalloproteinase 9 tissue inhibitor of metalloproteinases electroacupuncture histone acetylation

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