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Characteristic chemical profile of Juhe Fang extract with lipid-lowering properties

Kong JING ; Liu LULU ; Gao YUANYUAN ; Chen SIYU ; Li LINFU ; Shu YISONG ; Sun DAOHAN ; Jiang YANYAN ; Shi RENBING

Journal of Traditional Chinese Medical Sciences.2020;7(3):233-244.

Objective: The objective of this study was to verify the lipid-lowering effect of Juhe Fang extract (JHFE) and to determine its characteristic chemical profile in vitro and in vivo. Methods: A hyperlipidemia model was established by feeding mice a high-fat diet (HFD). After treatment for 30 days, serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were measured with an automatic biochemistry analyzer. The components from JHFE obtained from in vivo and in vitro experiments were investigated using an UPLC-Q Exactive-Orbitrap MS/MS. Results: The TC, TG, and LDL-C in the serum significantly decreased and the HDL-C significantly increased after JHFE treatment. A total of 95 compounds from JHEF including 15 phenolic acids (PA), 4 phenyl-ethanoid glycosides (PG), 24 flavonoids (F), 14 triterpenoids (T), 10 diterpenoid glycosides (D), 18 alka-loids (A) and 10 others (O) were identified. Trigonelline was discovered for the first time in a herbal medicine of Juhe Fang. Furthermore, 68 compounds were identified in vivo including 28 prototype compounds and 40 metabolites. The metabolic characteristics of these components were revealed including identification of new metabolites of 4-hydroxyphenyl ethyl-8-O-[α-L- arabinopyranosyl-(1→6)]-β-D-glucopyranoside (PEG) and lirinidine. A total of 43 components from JHFE were absorbed and/or metabolized. The contribution rate of each type of chemical component from JHFE to its lipid-lowering effect from high to low were A, F, PG, PA, D and T. Conclusion: The results of this study showed that JHFE demonstrated a significant lipid-lowering effect in a high-fat diet (HFD)-induced hyperlipidemia mouse model. Specific types of PA, PG, F, D, T and A formed the pharmaceutical architecture of the lipid-lowering effect of JHFE. This study should prove useful for clarifying the components responsible for the lipid-lowering effect of JHFE and provide a basis for precision quality control research.

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Effects of Fuhe decoction on behaviors and monoamine neurotransmitters in different brain regions of CUMS combined with social isolation depression model rats

Zheng RUOYUN ; Xiong WEIFENG ; He JUAN ; Wang XU ; Wang RANRAN ; Hao YU

Journal of Traditional Chinese Medical Sciences.2020;7(3):245-254.

Objective: To investigate the effect of Fuhe decoction on the behavior and levels of monoamine neuro-transmitters in different brain regions in a depression rat model induced by chronic unpredictable mild stimulation (CUMS) combined with social isolation.Methods: Fifty male SD rats were randomly divided into a blank group, model group, fluoxetine group, Chaiqinwendan decoction group, and Fuhe decoction group. Chronic unpredictable mild stimulation combined with a social isolation method was used to replicate the depression rat model. After 42 days of administration, a tail suspension test and high-performance liquid electrochemical detection (HPLC-ECD) were used to detect the behavioral changes and changes in the content of monoamine neurotransmitters norepinephrine (NE), dopamine (DA), 5-hydroxytrytamine (5-HT), and metabolites in different brain regions of rats in each group before and after treatment. Results: Compared with the model group, the epinephrine (E) content in the Fuhe decoction group was highly significantly increased (P < .01). Compared with the model group, the 5-HT content of the pre-frontal cortex in rats in the Fuhe decoction group was highly significantly increased (P < .01). Further-more, compared with the model group, the 5-HT content in the hippocampus of rats in the Fuhe decoction group was significantly increased (P<.05). Conclusion: Fuhe decoction can improve the depression-like behaviors of model rats, and its antide-pressant effect may be related to the increase in 5-HT content in the prefrontal cortex and hippocampus of rats.

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Pharmacological evaluation and mechanistic study of compound Xishu Granule in hepatocellular carcinoma

Li PIN ; Shi YUANYUAN ; Zhao BAOSHENG ; Xu WENHUI ; Xu ZIYING ; Zhang JINGXUAN ; Guo ZHAOJUAN ; Bi YUCONG ; Wang TIESHAN ; Qin YU ; Wang TING

Journal of Traditional Chinese Medical Sciences.2020;7(3):255-264.

Objective: In this study, we used HepG2 human hepatocellular carcinoma cells to study the effects of Compound Xishu Granule (CXG) on cell proliferation, apoptosis, and the cell cycle in vitro. We also used a xenograft tumor model to study the anti-tumor effects of CXG and related mechanisms in vivo.Methods: The effect of CXG on cell viability was measured using Cell Counting Kit-8 and a colony for-mation assay. The effect of CXG on apoptosis and the cell cycle was analyzed using flow cytometry. The in vivo anti-tumor effect of CXG was assessed by measuring the volume change in xenograft tumors after drug administration. The CXG anti-tumor mechanism was studied using western blotting assay to detect cell cycle and apoptotic associated proteins. Results: CXG suppressed HepG2 cell proliferation in a time-and dose-dependent manner in vitro. Colony formation experiments showed that CXG administration for 24 h significantly reduced HepG2 cell for-mations (P<.01). Flow cytometric analysis showed that CXG treatment for 48 h promoted apoptosis and blocked HepG2 cells in the G2/M phase. Western blotting results showed that Bax was significantly up-regulated and Bcl-2 was down-regulated in graft tumor tissues and HepG2 cells after CXG administra-tion, which increased the Bax/Bcl-2 ratio. PLK1, CDC25C, CDK1, and Cyclin B1 expression were up-regulated. CXG had a good inhibitory effect on graft tumor growth in vivo. Conclusion: CXG has good anti-tumor effects in vitro and in vivo. In vitro, CXG promoted HepG2 cell apoptosis and induced G2/M phase arrest. In vivo, CXG significantly inhibited graft tumor growth. The CXG mechanism in treating hepatocellular carcinoma may be that CXG can induce abnormal apoptotic and cell cycle associated protein expression, leading to mitotic catastrophe and apoptosis.

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Xiaoyaosan improves depressive-like behaviors by regulating the NLRP3 signaling pathway in the rat cerebral cortex

Chen CONG ; Yu RONG ; Xue ZHE ; Yan ZHIYI ; Bian QINLAI ; Hou YAJING ; Chen YUNZHI ; Liu YUEYUN ; Chen JIAXU

Journal of Traditional Chinese Medical Sciences.2020;7(3):265-273.

Objective: To observe changes in the molecular expression of the NLR Family Pyrin Domain Containing Protein 3 (NLRP3) pathway in depressed rats after treatment with Xiaoyaosan, and identify the regu-latory mechanism of this compound. Methods: Male Sprague-Dawley rats were randomly divided into five groups with 12 rats in each group, including the control group, model group, Fluoxetine group, Xiaoyaosan group, and MCC950 group. A depression model was generated by chronic immobilization stress (induced by 3 h of restraint immo-bilization every day), and the drugs were administered at the same time in each group for 21 days. The effects of Xiaoyaosan on behavioral changes of depressed rats were observed through macroscopic characterization, body mass, open field experiments, and a sucrose preference test. The mRNA and protein expression of the NLRP3 signaling pathway was examined by fluorescence real-time quantitative PCR and Western blot assays. Results: The Xiaoyaosan group, Fluoxetine group, and MCC950 group rats showed improved depressive behavior and an increased weight of sucrose water consumption. The protein and mRNA expression levels of NLRP3, Caspase-1, and IL-1β were also decreased in the Fluoxetine, Xiaoyaosan, and MCC950 groups. Conclusion: NLRP3, Caspase-1, and IL-1β protein and mRNA expression levels were increased in the cortex of depressed rats, while Xiaoyaosan protected cortical tissue in these rats by decreasing NLRP3, Caspase-1, and IL-1βprotein and mRNA expression.

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Protective effects of Buyinqianzheng Formula on mitochondrial morphology by PINK1/Parkin pathway in SH-SY5Y cells induced by MPP+

Ma HAOJIE ; Guo ZHENYU ; Gai CONG ; Cheng CUICUI ; Zhang JINKUN ; Zhang YUXIN ; Yang LUPING ; Feng WANDI ; Gao YUSHAN ; Sun HONGMEI

Journal of Traditional Chinese Medical Sciences.2020;7(3):274-282.

Objective: Buyinqianzheng Formula (BYQZF) is clinically employed in traditional Chinese medicine to treat Parkinson's disease (PD) by improving mitochondrial dysfunction. However, the underlying mechanisms by which BYQZF affects mitochondrial morphology remain unknown. Therefore, we observed the effects of BYQZF on mitochondria from the perspective of the PINK1/Parkin pathway. Methods: Cell survival rates were assessed by Cell Counting Kit-8 assay. Expression levels of PINK1 and Parkin mRNA were examined by qRT-PCR. Protein expression levels of PINK1, PINK1-Ser228, Parkin, Parkin-Ser65, Drp1, and Drp1-Ser637 were examined by western blotting. PINK1, Parkin, and Mito-Tracker? Red CMXRos (MTR) were stained by triple-labeled immunofluorescence, and observed under laser confocal microscopy. Results: Cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and protein expression levels of PINK1, Parkin, and Drp1-Ser637 were reduced after 1-methyl-4-phenylpyridinium (MPP+) intervention. In contrast, Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 increased significantly after MPP+intervention. Treatment with BYQZF increased cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and expression of PINK1, Parkin, and Drp1-Ser637 proteins. Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 decreased after BYQZF treatment. Conclusion: These results demonstrate that BYQZF has a protective effect on mitochondrial molecular mechanisms in the PD cell model, and the mechanism is related to the PINK1/Parkin pathway.

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Effect of the pineal gland on 5-hydroxytryptamine and γ- aminobutyric acid secretion in the hippocampus of male rats during the summer and winter

Li TING ; Wang HAILU ; Zhang HEWEI ; Liu LEILEI ; Li PEIPEI ; Ma SHURAN

Journal of Traditional Chinese Medical Sciences.2020;7(3):283-290.

Objective: The present study aimed to investigate the effect of seasonal variation on neurotransmitter release in the hippocampus of normal rats and rats with pineal excision. Methods: Two time points, the summer and winter solstice, which are the longest and shortest days of the year, respectively, were selected. Male Sprague-Dawley rats that underwent a sham operation without pineal excision were included as a control group. The concentrations of 5-hydroxytryptamine (5-HT) and γ-aminobutyric acid (GABA) were determined by radioimmunoassays and enzyme-linked immunosorbent assays, respectively. Results: In the winter, the 5-HT and GABA levels in normal rats exhibited a significant difference compared with those in the operation group (P < .01). A difference was also noted in GABA levels be-tween the normal group and the sham operation group (P<.05). The concentrations of 5-HT and GABA in the hippocampal tissues of the normal group exhibited a seasonal rhythm consisting of elevation during the summer and reduction during the winter (P < .01), while the GABA levels in the sham operation group exhibited a significant difference, with elevation during the summer and reduction during the winter (P<.01). In the operation group, GABA showed the same trend (P<.01). Conclusion: The seasonal rhythm of neurotransmitter secretion by the hippocampus (5-HT and GABA) consisted of elevation during the summer and reduction during the winter. During the winter, the pineal gland exhibited a reverse regulatory effect on the secretion of 5-HT and GABA in the hippocampus, and it exhibited seasonal selectivity with regard to the regulation of 5-HT.

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Anti-angiogenic effect of tripterygium glycosides tablets in animal models of rheumatoid arthritis: A systematic review and meta-analysis

Ao LIMEI ; Gao HAN ; Liu SHIMIN ; Jia LIFEN ; Liu BINGZHEN ; Guo JIE ; Liu JUN ; Dong QIUMEI

Journal of Traditional Chinese Medical Sciences.2020;7(3):291-300.

Objectives: To explore and summarize the beneficial effects of a traditional Chinese medicine prepara-tion, Tripterygium glycosides tablets (TGT), in rheumatoid arthritis (RA) animal models of neo-vascularization, and to provide a reference for future clinical applications and research on its pharmacologic mechanism.Methods: We searched the databases PubMed, Embase, Web of Science, Chinese National Knowledge Infrastructure, VIP, Wan Fang and SinoMed (China Biomedical Document Service System) to identify studies of TGT with outcome indicators of angiogenesis-related factors that were published before April 2020. Subgroup analysis and meta-regression were performed for dosage and duration of TGT. Statistical tests and subgroup analysis were conducted using RevMan 5.3, and meta-regression and sensitivity analysis were conducted using STATA/SE 15.0. Results: Fourteen studies of TGT in RA rats were included in this analysis. Treatment with TGT signifi-cantly reduces synovial microvessel density and the expression of vascular endothelial growth factor (VEGF), VEGF receptor 2, hypoxia inducible factor α, c-Fos, c-Jun, angiopoietin-1 and angiopoietin-2 compared with control groups (P < .05). Subgroup analysis did not show a significant association of the mRNA levels of VEGF in synovium, assessed using quantitative real-time PCR, with duration or dosage of TGT. Meta-regression analysis also indicated that the effects of dosage and duration were not significantly associated with differences in VEGF mRNA levels. Sensitivity analysis on VEGF mRNA levels did not fundamentally change the results. Conclusions: TGT can reduce synovial neovascularization by decreasing synovial microvessel density and expression of VEGF, VEGF receptor 2, hypoxia-inducible factorα, c-Fos, c-Jun, Ang-1 and Ang-2, thereby suppressing pannus formation and bone destruction in rat models of RA. Additional well-designed studies are required to confirm these findings.

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Evaluation of antidepressant activity of methanolic and hydroalcoholic extracts of Acorus calamus L. rhizome through tail suspension test and forced swimming test of mice

Yousuf SHAISTA ; Haq Marifatul SHIEKH ; Rasool AKHTAR ; Zulfajri MUHAMMAD ; Hanafiah Mohd MARLIA ; Nafees HUDA ; Tasneem SHOEIBA ; Mahboob MOHAMMED

Journal of Traditional Chinese Medical Sciences.2020;7(3):301-307.

Objective: Acorus calamus (AC) L. (Araceae) is an annual semi-aquatic and aromatic plant found in Europe, North America and Asia. Its rhizomes are often used by Native Americans, Americans, and Chinese as well as by other cultures. Ethnobotanical studies and documents have shown their use in various disease treatments, such as insomnia, mental disorders, diabetes mellitus, epilepsy, inflamma-tion, asthma, neuropathic pain, and diarrhea. In this study, the antidepressant activity of methanolic and hydroalcoholic extracts of the AC rhizome part in mice was investigated. Methods: Three doses of methanolic extract of AC rhizome (MEACR) (25, 50 and 100 mg/kg b.wt), three doses of hydroalcoholic extract of AC rhizome (HAACR) (100, 200 and 400 mg/kg b.wt), and standards (imipramine, 15 mg/kg b.wt and fluoxetine, 20 mg/kg b.wt) was daily oral administration to the mice for consecutive 14 days. The extract effect on the immobility time was monitored by a tail suspension test (TST) and a forced swimming test (FST). Monoamine oxidase (MAO) levels were also analyzed using standard methods. Results: The optimum antidepressant activity was viewed at 100 mg/kg b.wt of MEACR extract and 400 mg/kg b.wt of HAACR extract with 23.82％ and 20.59％ immobility period reduction, respectively. Besides, the extracts weakened the FST-induced elevation of MAO activity significantly and returned to near-normal levels of neurotransmitters in the brain. 100 mg/kg b.wt or above of MEACR extract significantly prevented the MAO-A and MAO-B activities in mice brain at a dose-dependent fashion. But, just 400 mg/kg b.wt of HAACR extract prevented the activity of MAO-A and MAO-B. Fluoxetine and imipramine showed a tendency to prevent the activity of MAO-A and MAO-B. Conclusion: This study suggests that AC rhizome extract mediated antidepressant activity by modulating the central neurochemical and hypothalamic-pituitary-adrenal (HPA) axis in response to FST and TST-induced stress. Therefore, AC rhizome extract can be used as a valuable plant supplement to treat depressive disorders.

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Mechanistic action of the acute toxicity of Bajitian (Morinda officinalis) in zebrafish embryos

Tan LIRONG ; Cheng MIN ; Gigi Chi Ting Auyeung ; Oscar Yu Suen Chan ; Chen XUEPING

Journal of Traditional Chinese Medical Sciences.2020;7(3):308-315.

Objectives: To investigate acute toxicity of Bajitian (Morinda officinalis) in zebrafish embryos. Methods: Zebrafish embryos at 48-h post fertilization (hpf) were exposed to Bajitian ethanol extract for 72 h. The causative action of a delay in yolk sac absorption by Bajitian was investigated by RT-PCR analysis of lipid metabolism-related microsomal triglyceride transfer protein (MTP), apolipoprotein C-Ⅱ(ApoC2) and lipogenesis-related liver x receptor (LXR) genes. The effect of Bajitian eliciting an in-flammatory response was studied by exposing 72 hpf myeloperoxidase (MPO):GFP transgenic zebrafish embryos to Bajitian extract for 4 h. Assessment was done by TUNEL, caspase-3/7, and RT-PCR analysis of the apoptosis related pathway B-cell lymphoma 2 associated X protein (Bax), Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) genes, neutrophil development-related stem cell leukaemia (SCL) and transcription factor PU.1 genes, to reveal the causative action of Bajitian reducing neutrophils. Results: RT-PCR analysis found that Bajitian extract had no effect on the expression of MTP or ApoC2 genes, but upregulated LXR gene, which might explain the delay in yolk sac absorption. Analysis of the inflammatory response showed that compared with negative controls, Bajitian extract significantly (P < .05) reduced the number of neutrophils in MPO: GFP embryos. TUNEL, caspase-3/7, and RT-PCR analysis of Bax and NF-kB genes found that Bajitian extract did not trigger the cell apoptosis. Further RT-PCR analysis found that Bajitian extract did not affect SCL expression, but did lead to down-regulation of PU.1. The inhibition of neutrophil development/differentiation may explain the decline in the total number of neutrophils following Bajitian treatment, which could be attributed to the anti-inflammatory effects found clinically for this drug. Conclusions: This study demonstrated that Bajitian caused a delay in yolk sac absorption and a decrease neutrophil in zebrafish embryos, which may be related to the inhibition of neutrophil development.

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Effect of cluster needling at scalp acupoints on differential protein expression in rat brain tissue after acute focal cerebral ischemia

Wu XIAONA ; Ni JINXIA ; An HUIYAN ; Gao YINTONG ; Li MIAOMIAO ; Huang ZHENZHEN ; Xu JINGNI

Journal of Traditional Chinese Medical Sciences.2020;7(3):316-324.

Objective: To explore the function of cluster needling at scalp points therapy on regulating differential protein's expression at different time points in middle cerebral artery occlusion (MCAO) model rats. Methods: Fifty-four rats were divided into three groups randomly and 18 rats in each group. The groups respectively were the model group (group M, n = 18), cluster needling at scalp points group (group C, n = 18), false operation group (group F, n = 18). Each group was then assigned in three subgroups, including 24-h, 7-day, and 14-day subgroups. Six rats in each subgroup. Acupuncture at Baihui (GV20) and 2 points beside Baihui, which was 3-4 mm away from the midline. Longa score was used to eval-uated neurological effects. Proteomics methods were used to identify differentially expression proteins with a standard of fold change greater than 1.5 and P<.05 at different times. Results: 1. Nerve function scoring: The nerve function scores at 7 and 14 days decreased in group C, which showed better neural function than group M (P<.05). 2. Fold change in proteins:Group M showed 932 differentially expressed proteins compared with group F, and among them, 414 proteins showed significant changes in expression after acupuncture. The expression levels of Cdc42 and GFAP were increased, and Mag, Shank2, and MBP levels were decreased. In the Gene Ontology analysis, the cellular component consisted of the terms cytoplasm, cytoskeleton, lysosome, and plasma membrane. The main related biological processes were cell-cell signaling, protein transport, aging, and cell adhesion. Many synaptic and metabolic pathways were found by KEGG analysis. Conclusion: Cluster needling at scalp acupoints can improve the nerve function score and improve dyskinesia in MCAO model rats. Cluster needling at scalp acupoints can regulate the expression of 414 proteins, including Cdc42, GFAP, Mag, Shank2, and MBP, which are related to cerebral ischemia. The differential proteins are major concentration in cytoplasm, cytoskeleton, lysosomes, and plasma mem-brane, participate in cell-cell signaling, protein transport, aging, and cell adhesion, and act through multiple synaptic and metabolic pathways to exert their biological functions.

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