期刊：
Journal of Ethnopharmacology,2024年323:117682 ISSN：0378-8741
通讯作者：
Liu, JJ;Wang, W
作者机构：
[Shi, Huan; Jiang, Lihong; Liu, Jianjun; Liao, Xiaolin; Zhao, Hongqing; Peng, Wangxia; Liu, JJ; Lu, Huaguan] Hunan Univ Tradit Chinese Med, Innovat Base Hunan State Key Lab Innovat Med & Tra, Sci & Technol Innovat Ctr, Changsha 410208, Peoples R China.;[Yang, Yupei; Wang, Wei] Hunan Univ Chinese Med, Innovat Mat Med Res Inst, Sch Pharm, TCM & Ethnomedicine Innovat & Dev Int Lab, Changsha 410208, Peoples R China.
通讯机构：
[Wang, W ; Liu, JJ ] H;Hunan Univ Tradit Chinese Med, Innovat Base Hunan State Key Lab Innovat Med & Tra, Sci & Technol Innovat Ctr, Changsha 410208, Peoples R China.;Hunan Univ Chinese Med, Innovat Mat Med Res Inst, Sch Pharm, TCM & Ethnomedicine Innovat & Dev Int Lab, Changsha 410208, Peoples R China.
摘要：
ETHNOPHARMACOLOGICAL RELEVANCE: The Kadsura coccinea (Lem.) A. C. Smith is known as "Heilaohu" of the Tujia ethnomedicine in China. It has anti-tumor, anti-oxidation, anti-HIV, anti-inflammatory and liver protective effects, used to treat diseases such as rheumatoid arthritis, cancer, gastritis and hepatitis. In this research, we investigated the anti-fibrotic effect and possible mechanisms of acetylbinankadsurin A (ACBA) in vitro and in vivo, which is a natural compound derived from roots of K. coccinea. AIM OF THE STUDY: We try to evaluate the efficacy of ACBA in the treatment of liver fibrosis and to explore the underlying mechanisms of ACBA by network pharmacology, machine learning, molecular docking, molecular dynamics simulations, and experimental assessment. MATERIALS AND METHODS: ACBA was isolated from the CH(2)Cl(2) layer of the roots of K. coccinea through column chromatographic techniques. The structure of ACBA was determined by using 1D and 2D NMR. CCl(4)-induced C57BL/6 mouse liver fibrosis models were established to evaluate the anti-fibrosis effects of ACBA in vivo. The molecular targets of ACBA and liver fibrosis were obtained from various databases, then constructed a protein-protein interaction (PPI) networks through the STRING database. Gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis were applied using the "clusterProfiler" R package. Furthermore, the key genes for ACBA treatment of liver fibrosis were identified by the least absolute shrinkage and selection operator (LASSO). Molecular docking and molecular dynamics simulations were also carried out. Finally, the target and pathway of ACBA were verified by immunofluorescence staining, RT-PCR and Western blot. RESULT: First, ACBA attenuated CCl(4)-induced liver injury and fibrosis in vivo. These findings were accompanied by decreased expression of α-SMA and collagen I. Second, ACBA significantly decreased serum levels of ALT, AST, TNF-α and IL-6. Then, we identified 133 potential targets of ACBA and 7987 targets of liver fibrosis. KEGG analysis showed that ACBA could regulate the drug metabolism - cytochrome P450, fructose and mannose metabolism, IL-17 and NF-κB signaling pathways. Next, six core targets was screened by LASSO analysis including AKR1B1, PFKFB3, EPHA3, CDK1, CCR1 and CYP3A4. Molecular docking showed that ACBA has a good binding affinity for CCR1. Furthermore, compared with CCR1 inhibitor BX-471, The results of molecular simulation dynamics showed that ACBA was stable in binding with CCR1. Consistently, ACBA remarkably downregulated the expression of CCR1, p-NF-κBp65, p-IκBα, p-STAT1 and TNF-α proteins, which were upregulated in CCl(4)-induced hepatic fibrosis and LPS-THP-1cells. CONCLUSION: Our results suggest that ACBA significantly attenuated CCl(4)-induced liver fibrosis in histopathological and in serum level. This effect may be mediated by CCR1, NF-κB and STAT1 signalling.
作者机构：
[Yuan Ma; Lijuan Zhao] School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China;These authors contributed equally to this work.;Chongqing Innovation Institute, China Pharmaceutical University, Chongqing 401135, China;State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China;[Qianqian Wu; Jianping Kong] School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China
通讯机构：
[Juan Li; Shunxiang Li] H;Hunan Province Sino-US International Joint Research Center for Therapeutic Drugs of Senile Degenerative Diseases, Changsha 410208, China<&wdkj&>Authors to whom correspondence should be addressed.<&wdkj&>School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China<&wdkj&>Hunan Engineering Technology Research Center for Bioactive Substance Discovery of Chinese Medicine, Changsha 410208, China
摘要：
COVID-19, caused by SARS-CoV-2, has emerged as the most destructive emerging infectious disease of the 21st century. Vaccination is an effective method to combat viral diseases. However, due to the constant mutation of the virus, new variants may weaken the efficacy of vaccines. In the current field of new coronavirus research, viral protease inhibitors have emerged as a highly regarded therapeutic strategy. Nevertheless, existing viral protease inhibitors do not fully meet the therapeutic needs. Therefore, this paper turned to traditional Chinese medicine to explore new active compounds. This study focused on 24 isolated compounds from Acorus calamus L. and identified 8 active components that exhibited significant inhibitory effects on SARS-CoV-2 PLpro. Among these, the compound 1R,5R,7S-guaiane-4R,10R-diol-6-one demonstrated the best inhibitory activity with IC50 values of 0.386 ± 0.118 μM. Additionally, menecubebane B and neo-acorane A exhibited inhibitory activity against both Mpro and PLpro proteases, indicating their potential as dual-target inhibitors. The molecular docking results confirmed the stable conformations of these compounds with the key targets and their good activity. ADMET and Lipinski’s rule analyses revealed that all the small molecule ligands possessed excellent oral absorption properties. This study provides an experimental foundation for the discovery of promising antiviral lead compounds.
摘要：
Objective: To investigate the effect of isorhamnetin on the pathology of rheumatoid arthritis (RA). Methods: Tumor necrosis factor (TNF)- alpha -induced fibroblast-like synoviocytes (FLS) was exposed to additional isorhamnetin (10, 20 and 40 mu mol/L). Overexpression vectors for matrix metalloproteinase-2 (MMP2) or MMP9 or SRC were transfected to explore their roles in isorhamnetin-mediated RA-FLS function. RA-FLS viability, migration, and invasion were evaluated. Moreover, a collagen-induced arthritis (CIA) rat model was established. Rats were randomly divided to sham, CIA, low-, medium-, and high-dosage groups using a random number table (n=5 in each group) and administed with normal saline or additional isorhamnetin [2, 10, and 20 mg/(kg<middle dot>day)] for 4 weeks, respectively. Arthritis index was calculated and synovial tissue inflammation was determined in CIA rats. The levels of MMP2, MMP9, TNF-alpha, interleukin-6 (IL-6), and IL-1 beta, as well as the phosphorylation levels of SRC, extracellular regulated kinase (ERK), and cyclic adenosine monophosphate response element-binding (CREB), were detected in RA-FLS and synovial tissue. Molecular docking was also used to analyze the binding of isorhamnetin to SRC. Results: In in vitro studies, isorhamnetin inhibited RA-FLS viability, migration and invasion (P<0.05). Isorhamnetin downregulated the levels of MMP2, MMP9, TNF-alpha, IL-6, and IL-1 beta in RA-FLS (P<0.05). The overexpression of either MMP2 or MMP9 reversed isorhamnetin-inhibited RA-FLS migration and invasion, as well as the levels of TNF-alpha, IL-6, and IL-1 beta (P<0.05). Furthermore, isorhamnetin bound to SRC and reduced the phosphorylation of SRC, ERK, and CREB (P<0.05). SRC overexpression reversed the inhibitory effect of isorhamnetin on RA-FLS viability, migration and invasion, as well as the negative regulation of MMP2 and MMP9 (P<0.05). In in vivo studies, isorhamnetin decreased arthritis index scores (P<0.05) and alleviated synovial inflammation. Isorhamnetin reduced the levels of MMP2, MMP9, TNF-alpha, IL-6, and IL-1 beta, as well as the phosphorylation of SRC, ERK, and CREB in synovial tissue (P<0.05). Notably, the inhibitory effect of isorhamnetin was more pronounced at higher concentrations (P<0.05). Conclusion: Isorhamnetin exhibited anti-RA effects through modulating SRC/ERK/CREB and MMP2/MMP9 signaling pathways, suggesting that isorhamnetin may be a potential therapeutic agent for RA.
期刊：
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY,2024年169:106550 ISSN：1357-2725
通讯作者：
Wang, Wei;Yu, Huanghe
作者机构：
[Shen, Xinyang; Li, Huanjie; Sheng, Wenbing; Li, Yunzhe; Deng, Yasi; Tian, Xing; Lin, Haokai; Yuan, Hanwen; Zheng, Hao; Li, Bin; Yang, Yong] TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Materia Medica Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China;[Wang, Wei] TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Materia Medica Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China. Electronic address: wangwei402@hotmail.com;[Yu, Huanghe] TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Materia Medica Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China. Electronic address: yhh@hnucm.edu.cn
通讯机构：
[Wang, Wei; Yu, Huanghe] T;TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Materia Medica Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China. Electronic address:
摘要：
Tujia ethnomedicine Xuetong (the stems of Kadsura heteroclita) have been widely used in folk for rheumatoid arthritis (RA), which can alleviate rheumatic pain through liquor soaking in folk. In this study, we aimed to evaluate the pharmacological effects and underlying mechanism of Xuetongsu (a key chemical component of Xuetong) on bone destruction. In our previous study, it was found that Xuetong extract can reduce adjuvant arthritic rats paw swelling and inhibit inflammatory factors in serum. Furthermore, Xuetongsu has been demonstrated to inhibit the proliferation of fibroblast-like synoviocytes, but its potential to inhibit bone destruction has not been explored. To address this, we employed the STRING database to predict protein interactions and utilized Autodock software to simulate the binding of Xuetongsu to target proteins. In this study, administration of Xuetongsu significantly alleviated paw swelling and bone destruction in C57BL/6 mice with collagen-induced arthritis (CIA). Mechanistic studies have indicated that Xuetongsu promotes apoptosis of mature osteoclasts in joint tissues by activating Caspase-3 and Bax, while inhibiting Bcl-2. Additionally, Xuetongsu inhibits osteoclast differentiation by suppressing RANKL, RANK, P-NF-κB, and NFATc1, and reduces bone resorption activity by inhibiting MMP-9, CTSK, and TRAP. Importantly, Xuetongsu exhibits good biocompatibility in major organs of mice. In summary, Xuetongsu has the potential to treat bone destruction by promoting apoptosis of mature osteoclasts, inhibiting osteoclast differentiation, and reducing bone resorption. This study reveals the pharmacological effects of Xuetongsu and its mechanism of action, which may contribute to the development of novel approaches for treating RA.
作者机构：
[Wei Su; Huanghe Yu; Qingling Xie; Ling Liang; Hanwen Yuan; Mengying Lyu; Bin Li] TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Material Medical Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China;[Wei Wang] Author to whom correspondence should be addressed.
通讯机构：
[Wei Wang] A;Author to whom correspondence should be addressed.
摘要：
Xuetong, the dried stem of Kadsura heteroclita (Roxb.) Craib, is a traditional Tujia medicine extensively used to treat rheumatoid arthritis (RA). All traditional Chinese medicines (TCMs) necessitate a processing stage called “Paozhi” before clinical application. However, there is a dearth of research concerning the processing methods employed for Xuetong. To investigate the impact of vinegar and wine processing on the chemical constituents of Xuetong, this study devised a targeted offline two-dimensional (2D) high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography-orbitrap mass spectrometry (UHPLC-orbitrap-MS) method. By incorporating various MS data-processing techniques, such as molecular networking technology, fragment-ion similarity searching (FISh), online and offline database matching, and fragmentation pattern analysis, a total of 158 components were identified in Xuetong. Among them, 14 were verified by comparison with the reference standards. Notably, aside from triterpenoids and lignans, catechin derivatives were found to be the predominant constituents of Xuetong, and their levels exhibited a significant decrease following processing. This method significantly improved peak capacity and resolution, overcoming the limitations of 1D LC in simultaneously analyzing highly polar catechin derivatives and less polar triterpenoids and lignans. Moreover, the developed method shows promise for Xuetong’s quality control.
作者机构：
[Fu, Fuhua; Mao, Yingchao; Zhu, Lingfeng] Hunan Acad Agr Sci, Hunan Agr Prod Proc Inst, Changsha 410125, Peoples R China.;[Chen, Jiajing; Huang, Dan; Liu, Jing; Lei, Chang; Zhu, Lijun] Hunan Univ Chinese Med, Sci & Technol Innovat Ctr, State Key Lab Chinese Med Powder & Med Innovat Hun, Changsha 410208, Peoples R China.
通讯机构：
[Huang, D; Lei, C ] H;Hunan Univ Chinese Med, Sci & Technol Innovat Ctr, State Key Lab Chinese Med Powder & Med Innovat Hun, Changsha 410208, Peoples R China.
摘要：
Puerariae Radix is one of the most widely used ancient traditional Chinese medicines and is also consumed as food, which has rich edible and medicinal value. Puerariae Radix can be divided into Puerariae Lobatae Radix (PL) and Puerariae Thomsonii Radix (PT). These two medicinal materials are very similar, and they are often mixed up or misused. In this study, gas chromatography-ion migration spectrometry (GC-IMS) was used to analyze the volatile organic compounds (VOCs) of PL and PT, and the differences in VOCs were analyzed using fingerprint, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). The results showed that a total of 173 VOCs were obtained from PL and PT, and 149 were qualitatively identified, including 38 aldehydes, 22 alcohols, 22 ketones, 19 esters, 13 esters, 10 acids, 10 pyrazines, 6 terpenes, 4 furans, and 2 pyridines. The characteristic VOCs of PL and PT were clarified by constructing GC-IMS fingerprints. PL and PT can be effectively distinguished, and five characteristic VOCs were screened using PCA and OPLS-DA analysis methods. This study identified and evaluated the types and differences in VOCs in PL and PT. The established method is simple, rapid, accurate, and sensitive, and it provides theoretical guidance for the identification, tracing, and quality evaluation of PL and PT.
期刊：
Journal of Affective Disorders,2024年348:107-115 ISSN：0165-0327
通讯作者：
Chen, Naihong;Yang, Yantao;Yang, Songwei
作者机构：
[Ai, Qidi; Lin, Yuting; Yao, Jiao; Liu, Xinya; Tian, Zhifeng; Sun, Yang; Wang, Huiqin; Lin, Meiyu; Yang, Songwei; Yan, Qian; Yang, Yantao; Long, Junpeng] Hunan Engineering Technology Center of Standardization and Function of Chinese Herbal Decoction Pieces, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China;[Ai, Qidi; Lin, Yuting; Yao, Jiao; Liu, Xinya; Tian, Zhifeng; Sun, Yang; Wang, Huiqin; Lin, Meiyu; Yan, Qian; Long, Junpeng] Key Laboratory of Modern Research of TCM, Education Department of Hunan Province, Changsha 410208, China;[Chen, Cong] School of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;[Gao, Yan] Biomedical Innovation Center, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China;[Chen, Naihong] State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica & Neuroscience Center, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100050, China. Electronic address: chennh@imm.ac.cn
通讯机构：
[Chen, Naihong] S;[Yang, Yantao; Yang, Songwei] K;State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica & Neuroscience Center, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100050, China. Electronic address:;Key Laboratory of Modern Research of TCM, Education Department of Hunan Province, Changsha 410208, China. Electronic address:
摘要：
BACKGROUND: Depression is a refractory psychiatric disorder closely associated with dysfunction of the gap junctions (GJs) between astrocytes as well as neuroinflammation. Higenamine (Hig) is a potent cardiotonic ingredient in Fuzi (i.e., Aconitum carmichaeli Debx.) with anti-inflammatory and antioxidant effects, which has a significant protective effect on damaged nerve cells and has great potential for the treatment of neuropsychiatric diseases. METHODS: Rats were stimulated by chronic unpredictable stress (CUS) for 28days while given Hig (5, 10, 20mg/kg) and then analyzed behaviorally by the open field test, sucrose preference test, and forced swimming test. Changes in astrocyte GJs function and morphology were observed by dye transfer and transmission electron microscopy, respectively. Expression and phosphorylation of connexin 43 (Cx43) were analyzed by Western blot. Also, considering the close relationship between depression and neuroinflammation, we determined the inflammatory response in serum with ELISA kits and analyzed the expression of inflammation-related proteins with Western blot. RESULTS: Hig ameliorated CUS-induced depression-like behavior in rats. Hig administration improved gap junctional dysfunction in astrocytes, reduced gap junctional gaps and elevated the expression of Cx43 and decreased the phosphorylation of Cx43. Meanwhile, Hig administration was also able to attenuate the inflammatory response that occurs after CUS in rats. LIMITATIONS: For the role of Cx43 in depression, we did not validate it more deeply in animal models with knockout Cx43. In addition, GJs dysfunction might be associated with the inflammatory response seen in depression, but this needs to be further investigated. CONCLUSIONS: Hig ameliorates depression and exerts its antidepressant effect possibly by improving the dysfunctional GJs between astrocytes and the inflammatory response.
期刊：
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY,2024年34(2):467-475 ISSN：1017-7825
作者机构：
[Gao, Xin; Yuan, Xiwei; Tan, Chaoyang; Xu, Dehong; Wu, Wenmei] Biological Engineering Laboratory, College of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, P.R. China;[Xu, Dehong; Li, Shunxiang] Hunan Engineering Technology Research Center for Bioactive Substance Discovery of Chinese Medicine, Changsha, Hunan 410208, P.R. China;[Xu, Dehong; Li, Shunxiang] Hunan Province Sino-US International Joint Research Center for Therapeutic Drugs of Senile Degenerative Diseases, Changsha, Hunan 410208, P.R. China
摘要：
ρ-Hydroxyacetophenone is an important and versatile compound that has been widely used in medicine, cosmetics, new materials, and other fields. At present, there are two ways to obtain ρ-hydroxyacetophenone. One is to extract it from plants, such as Artemisia capillaris Thunb and Cynanchum otophyllum Schneid, and the other is to synthesize it by using chemical methods. Of these two methods, the second is the main one, although it has problems, such as flammable and explosive reagents, difficult separation of by-products, and harsh reaction conditions. To solve these issues, we adopted genetic engineering in this study to construct engineered Escherichia coli containing Hped gene or EbA309 gene. Whole-cell biotransformation was conducted under the same conditions to select the engineered E. coli with the higher activity. Orthogonal tests were conducted to determine the optimal biotransformation condition of the engineered E. coli. The results showed that the optimal condition was as follows: substrate concentration of 40 mmol/l, IPTG concentration of 0.1 mmol/l, an induction temperature of 25°C, and a transformation temperature of 35°C. Under this condition, the effects of transformation time on the ρ-hydroxyacetophenone concentration and cell growth were further studied. We found that as the transformation time extended, the ρ-hydroxyacetophenone concentration showed a gradually increasing trend. However, when the ρ-hydroxyacetophenone concentration increased to 1583.19 ± 44.34 mg/l in 24 h, cell growth was inhibited and then entered a plateau. In this research, we realized the synthesis of ρ-hydroxyacetophenone by biotransformation, and our findings lay a preliminary foundation for further improving and developing this method.
作者机构：
[Dai, Jie; Huang, Shiyi; Huang, Chencun; Zhao, Hongqing; He, Xiangchang; He, Weihe; Xu, Guangming] School of Pharmacy, Hunan University of Chinese Medicine, Changsha, China
关键词：
Polygonum amplexicaule D. Don var. sinense Forb;affinity ultrafiltration mass spectrometry;molecular docking;thrombin;zebra fish
摘要：
INTRODUCTION: Polygonum amplexicaule D. Don var. sinense Forb (PAF), a medicinal plant, has the effect of promoting blood circulation and removing blood stasis. However, the active compounds and targets of its anticoagulant effect are still unclear. OBJECTIVES: This study aims to establish an effective reversely thrombin-targeted screening method for anticoagulant active components in PAF by affinity ultrafiltration (AUF) coupled with ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectroscopy (UPLC-Q-TOF-MS). METHODS: Different polar parts of PAF were screened for potential thrombin ligands by AUF-HPLC and identified by UPLC-Q-TOF-MS. After studying the affinity between ligands and thrombin by molecular docking, the antithrombotic activity of ligands was detected in vivo by zebrafish thrombus model, and in vitro by chromogenic substrate method. The mechanism of such ligands on thrombin was further studied by coagulation factor assay. RESULTS: Eleven potential thrombin ligands from PAF were screened by the AUF-UPLC-Q-TOF-MS method, and two compounds (butyl gallate and β-sitosterol) with significant anticoagulant activity were discovered via in vitro and in vivo activity testing. CONCLUSION: A method system based on AUF-UPLC-Q-TOF-MS, molecular docking and in vivo and in vitro experiments also provided a powerful tool for further exploration of anticoagulant active components in PAF.
