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Objective:To explore the material basis and mechanism of Linggui Zhugan Decoction in treating hypertension and obesity by means of network pharmacology and molecular docking technique.Methods:The TCMSP was retrieved and the main active components and action targets of Linggui Zhugan Decoction were screened. The GeneCards, OMIM, TTD, DisGeNET and DrugBank databases were used to screen disease-related targets of hypertension and obesity. The Cytoscape 3.9.0 was used to draw Chinese materia medica-composition-intersection target-disease network diagram. The STRING 11.5 database was used to draw PPI network. The cytoNCA plug-in was used to screen core active components and targets. The bioenrichment analysis of GO and KEGG was carried out in the R4.1.2, and the Chinese materia medica-intersection target-path diagram was drawn, and the core active components and core targets were docked in PyMOL and AutoDockTools 1.5.7.Results:A total of 102 potentially active components and 62 intersection targets were obtained, and 8 active components and 7 core targets were screened. Enrichment analysis showed that the key targets were mainly enriched through the signaling pathways of fluid shear stress and atherosclerosis, lipid and atherosclerosis, and AGE-RAGE, which were involved in biological processes such as the response to nutritional levels and the regulation of small molecule metabolism. Molecular docking showed that there were 37 groups with addinity < -7 kcal/mol.Conclusion:The main active components of Linggui Zhugan Decoction are quercetin, kaempferol and naringenin, which may play a role in fluid shear stress and atherosclerosis pathway, lipid and atherosclerosis pathway and AGE-RAGE signal pathway through AKT1, EGFR, IL1B and other targets.
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OBJECTIVE To explore the enhancement effect of Linggui zhugan decoction （LGZG） regulating autophagy on doxorubicin （DOX） against non-alcoholic fatty liver disease-hepatocellular carcinoma （NAFLD-HCC）. METHODS C57BL/6 mice were randomly divided into blank group， NAFLD-HCC group， LGZG group， DOX group and DOX+LGZG group， with 10 mice in each group. The NAFLD-HCC model was established by intraperitoneal injection of diethylnitrosamine （50 mg/kg） and high-fat diet. The blank group was injected with the same amount of normal saline and fed with ordinary diet. After modeling， administration groups were given LGZG aqueous extract （20 g/kg） intragastrically and/or DOX solution intraperitoneally （8 mg/kg）； the blank group and NAFLD-HCC group were given a constant volume of normal saline intragastrically， once a day， for 4 consecutive weeks. The general condition of mice （No.2022-BS-197） was monitored during modeling and drug intervention. After drug intervention， body weight， liver weight and liver coefficient of mice were detected. The histopathologic morphology and fibrosis degree of liver tissue in mice were observed； the levels of blood lipid [the levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C）]， the serum contents of alpha fetoprotein （AFP） and carcinoembryonic antigen （CEA）， and the expressions of marker of proliferation Ki-67， B-cell lymphoma-2 （Bcl-2） and Bcl-2 associated X （Bax） in liver tissue were all detected as well as protein expressions of microtubule-associated proteins 1A and 1B （LC3）， Beclin1 and selective autophagy adopt proteins P62. RESULTS Compared with the blank group， the activity of mice decreased gradually as time in the NAFLD-HCC group； mental fatigue， disheveled and matte hair were observed， and body weight decreased significantly （P＜0.05）； liver weight had an upward trend， and liver coefficient increased significantly （P＜0.05）. The inflammatory cells of liver tissue were infiltrated， with some cells showing ballooning and small cell hyperplasia， and the degree of liver fibrosis was worsened； serum levels of TC， TG and LDL-C， AFP and CEA contents increased significantly， while HDL-C level decreased significantly （P＜0.05）. The protein expressions of Ki-67 and Bcl-2 in liver tissue were increased significantly （P＜0.05）， while the protein expression of Bax decreased. The protein expression of Beclin1 in liver tissue decreased significantly （P＜0.05）； LC3Ⅱ/ LC3Ⅰ decreased， while the expression of P62 protein increased. Compared with the NAFLD-HCC group， the above indexes of mice were improved to different extents in the DOX group， LGZG group and DOX+LGZG group， and the intervention effect of DOX combined with LGZG were better than those of DOX. CONCLUSIONS LGZG combined with DOX can synergically promote the apoptosis of tumor cells， enhance the sensitivity of NAFLD-HCC chemotherapy， and effectively slow down the occurrence and development of NAFLD-HCC. The mechanism may be related to the regulation of autophagy in tumor cells.
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To verify the appropriate preparation process of extracts for the solid substance benchmark of Linggui Zhugan Decoction. The extracts were prepared by different preparation processes, namely the traditional process(process 1), the extract combined with volatile oil separated from traditional process extract liquid(process 2), the modern secondary reflux extraction process(process 3) and the process that volatile oil was extracted first, then prepared according to the traditional process, and combined with extract(process 4); based on the characteristic spectrum, index components of cinnamaldehyde, glycyrrhizin, ammonium glycyrrhizinate, cinnamic acid, and the dry extract rate of process 1, the differences and similarities of four extracts were compared. The results showed that the similarity of the characteristic spectrum of process 2, process 4 and process 1 were all greater than 0.97, while there was no significant difference for the content of 4 quality control components and dry extract rate; the similarity of the characteristic spectrum of process 3 and process 1 was 0.91, the absolute peak area of 13 out of 21 peaks and the relative peak area of 7 peaks increased significantly, and the content of 3 out of 4 quality control components and dry extract rate also significantly increased. In conclusion, the material standards of extracts by the process 2 and 4 are consistent with that of the traditional process, so the two processes are suitable.
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Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Glycyrrhizic Acid , Oils, Volatile , Quality Control , Reference StandardsABSTRACT
OBJECTIVE@#To investigate the inhibitory effect of Linggui Zhugan Decoction (LZD, ) on the ventricular remodeling (VR) after acute myocardial infarction (AMI) and related mRNA and proteins expression in transforming growth factor-beta 1 (TGF-β)/Smad signaling pathway, and explain its putative mechanism.@*METHODS@#A VR model was generated by ligation of coronary artery in mice. Two weeks after surgery, 60 mice were randomly divided into the model group, the sham-operation group (distilled water), the positive control group (2.4 mg/kg simvastatin), and the low-, medium- and high-dose LZD groups (2.1, 4.2, 8.4 g crude drug/kg, respectively) by a random number table, 10 mice in each group. Mice in each group was treated for 4 weeks. Changes of hemodynamics indices and cardiac weight index were detected by the PowerLab data acquisition and analysis recording instrument. Morphology changes of myocardial tissue were observed by hematoxylin-eosin and Masson staining. The expressions of TGF-β, Smad2, Smad3, p-Smad2 and p-Smad3 in myocardial tissue were detected by Western blotting. The mRNA expressions of TGF-β, Smad2 and Smad3 were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The expressions of matrix metalloprotein 2 (MMP2), MMP9, collagen I and collagen III were observed by immunohistochemical methods.@*RESULTS@#VR mice showed significant dysfunction in hemodynamic indices and cardiac structure and function. Compared with the shamoperation group, myocardial tissue damage, interstitial fibrosis occurred in the model mice, left ventricular systolic pressure (LVSP), left ventricular pressure maximum contraction rate (+dp/dt) and left ventricular pressure maximum relaxation rate (-dp/dt) decreased significantly (all P<0.01), while left ventricular end-diastolic pressure (LVEDP), cardiac weight index and left ventricular weight index elevated significantly, meanwhile TGF-β, p-Smad2, p-Smad3, Smad2, Smad3, MMP2, MMP9, collagen I, collagen III protein expressions in myocardial tissue and TGF-β, Smad2 and Smad3 mRNA expressions increased significantly (all P<0.01). Compared with the model group, LZD could significantly improve the pathological changes of myocardial tissue, increase LVSP, +dp/dtmax and -dp/dtmax, lower LVEDP, reduce the whole heart weight index and left ventricular weight index and inhibit the over-expressions of TGF-β, p-Smad2, p-Smad3, Smad2, Smad3, MMP2, MMP9, collagen I and collagen III proteins in myocardial tissue and mRNA expressions of TGF-β, Smad2 and Smad3 (P<0.05 or P<0.01).@*CONCLUSION@#LZD can significantly suppress VR induced by AMI, and its underlying mechanism may be associated with its inhibitory effect on the TGF-β1/Smad signaling pathway.
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Animals , Male , Disease Models, Animal , Myocardial Infarction , Plant Extracts , Pharmacology , Rats, Sprague-Dawley , Smad Proteins , Metabolism , Transforming Growth Factor beta1 , Metabolism , Ventricular RemodelingABSTRACT
Objective: To establish an HPLC fingerprint of classical named Linggui Zhugan Decoction and determination method of the content of three compounds, and provide reference for the study of the material basis of the classical Linggui Zhugan Decoction. Methods: The method was performed by high performance liquid chromatography with CNW Athena C18 (250 mm × 4.6 mm, 5 μm) and gradient elution with acetonitrile (A)-0.1% phosphoric acid (B) as the mobile phase, gradient elution: 0-10 min, 5%-10%; 10-20 min, 10%-15% A; 20-50 min, 15%-30% A; 50-75 min, 30%-40% A; 75-95 min, 40%-50% A; 95-111 min, 50%-95% A. The flow rate was 1.0 mL/min, the injection volume was 10 μL, and the column temperature is 30 ℃. The fingerprint and the detection wavelength of glycyrrhizin and ammonium glycyrrhizinate were 230 nm, and cinnamic aldehyde was 290 nm. The chromatographic fingerprint evaluation system published by the State Pharmacopoeia Commission (2012 Edition) was used to establish the fingerprint of 10 batches of Lingguizhugan decoction, and the content of three kinds of index components were determined simultaneously by the established HPLC method. Results: The research on the 10 batches of Lingguizhugan soup showed that the fingerprint similarity was greater than 0.9, and 24 common peaks were calibrated, and the peak resolution was good. The content determination results showed that the content of glycyrrhizin and ammonium glycyrrhizinate was high. According to the methodological investigation, the three components showed a good linear relationship within a certain concentration range; The precision RSD values were all less than 1.0%; The average recovery rates of glycyrrhizin, cinnamaldehyde and ammonium glycyrrhizinate were 98.35%, 101.51%, and 102.59%, respectively. The RSD is less than 2.5%; The sample is stable within 48 h, and the method has good repeatability. Conclusion: The fingerprint method and the determination method of three components in the traditional Chinese medicine classical prescription named Linggui Zhugan Decoction established in this study are simple, stable, accurate and reliable, and can be used for the quality control of the Linggui Zhugan Decoction.
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Objective: To study the potential mechanism of Linggui Zhugan Decoction in the treatment of Alzheimer' s disease (AD) by using network pharmacological technology. Methods: Firstly, TCMSP, Swiss, SuperPred, Stitch databases and supplement literatures were combined to search for the chemical compounds and the potential targets related to the four herbs in Linggui Zhugan Decoction, using the oral bioavailability (OB) ≥ 30%, drug-like (DL) ≥ 0.18 as screening conditions. Meanwhile, the targets related to AD were screened through OMIM, TTD, GAD and Pharm Gkb databases. Moreover, the interactive targets of Linggui Zhugan Decoction and AD were further acquired. Then, Gene Ontology (GO) analysis and KEGG metabolic pathway enrichment analysis of interactive targets were performed using DAVID 6.8 database. Finally, degree and betweenness centrality analysis were performed to find potential key targets and key pathways after the "Herbs-Active Compounds-Targets" interaction network was constructed by Cytoscape software. Results: There were 126 potential active compounds and 120 potential interactive targets in Linggui Zhugan Decoction. A total of 565 enrichment results were obtained by GO analysis, including 396 biological processes, 121 molecular functions and 48 cell components, mainly involving protein binding, cell, oxidation-reduction process, etc. Simultaneously, the selected targets were enriched by KEGG analysis and it indicated that 67 metabolic pathways play an important role in AD, primarily including neuroactive ligand-receptor interaction, pathways in cancer, serotonergic synapse and calcium signaling pathway, etc. Conclusion: The synergetic effect of Linggui Zhugan Decoction with multi-compounds, multi-targets and multi-pathways in the treatment of AD were revealed by network pharmacology, which provides a theoretical basis for the study of its pharmacodynamic material basis and action mechanism.
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Objective To explore the effect of Linggui Zhugan decoction on the level of serum inflammatory factors , blood lipid and oxidation with coronary heart disease (CHD).Methods 123 cases of patients diagnosed as CHD and met criterions were collected from May 2013 to February 2015 in cardiovascular department of our hospital.They were divided into two groups according to different treatment modalities, observation group and control group.General information of the two groups were comparable, The patients of two groups reveived conventional treatments such as taking aspirin enteric-coated tablets, rosuvastatin, nitrate.The patients of observation group received Linggui Zhugan Decoction on the basis of conventional treatments.The serum levels of TC, TG, LDL-C, HDL-C, TNF-α, IL-6, IL-1β, SOD, MDA were recorded before and after treatment.Results The serum levels of TC, TG, LDL-C of observation groups were lower than those of control group after treatment (P<0.05).And the serum levels of HDL-C of observation groups were higher than those of control group after treatment (P<0.05).There was significant improvement in the serum levels of TNF-α, IL-6, IL-1β, MDA and SOD of two groups after treatment than those before treatment (P<0.05).The serum levels of TNF-α, IL-6, IL-1β, MDA of observation group were lower than those of control group after treatment ( P<0.05 ) , while the SOD serum levels of observation group were higher than those of control group after treatment (P<0.05).Conclusion Linggui Zhugan decoction could significantly improve the levels of blood lipids and decrease the levels of inflammation and oxidative stress in patients of CHD with significant effect.
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Objective To observe the effectiveness and safety of Linggui Zhugan Decoction in the treatment of metabolic syndrome(MS) induced by antipsychotics.Methods Sixty MS patients were enrolled who also matched the diagnostic criteria for phlegm stagnation type.These 60 patients who were treated in the Second Affiliated Hospital of Xinxiang Medical University were randomly divided into the observation group and control group,each group included 30 patients.The patients in the observation group were given the treatment of Linggui Zhugan Decoction in addition to antipsychotic treatment,while the patients in the control group were only given antipsychotic treatment.After two weeks or four weeks of treatment,the levels of triglyceride (TG),high density lipoprotein (HDL),fasting plasma glucose,insulin,C-peptide and insulin resistance index (HOMA-IR) were compared between the two groups before and after treatment.Results After two weeks or four weeks treatment,the levels of TG,fasting plasma glucose,insulin,C-peptide and HOMA-IR significantly increased and that of HDL decreased significantly in the control group patients compared with those before treatment (P ＜ 0.05),while the results in the observation group were opposite (P ＜ 0.05).In the control group patients,the levels of insulin,HOMA-IR and C-peptide after four weeks treatment were significantly higher than those after two weeks treatment (P ＜ 0.05).However,in the observation group patients,the levels of insulin,HOMA-IR and C-peptide after four weeks treatment were significantly lower than those after two weeks treatment (P ＜ 0.05).Compared with the control group,after two weeks or four weeks treatment the levels of TG,fasting plasma glucose,insulin,C-peptide and HOMA-IR decreased significantly in the observation group patients and that of HDL increased significantly (P ＜ 0.05).The treatment emergent symptom scale score of the observation group was lower than that of the control group at the end of four weeks (P ＜ 0.05).Conclusion Linggui Zhugan Dcoction can improve MS caused by antipsychotics and has good safety.
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Objective To approach the therapeutic effect of modified Linggui Zhugan decoction for treatment of patients with type Ⅱ cardiorenal syndrome (CRS).Methods Thirty patients with CRS admitted to Affiliated Hospital of Tianjin Institute of Chinese Medicine were selected. By a random number table and double-blind method, they were divided into two groups: treatment and control groups, 15 cases in each group. The patients in both groups were treated with conventional western medicine, and those in the treatment group were additionally given traditional Chinese medicine (TCM) modified Linggui Zhugan decoction (including the following ingredients: Poria 30 g, Cinnamomi Cortex 10 g, Atractylodis Macrocephalae Rhizoma 15 g, Glycyrrhizae Radix et Rhizoma Preparata cum Melle 10 g, Morindae Officinalis Radix 15 g, Arecae Pericarpium 30 g, Astragali Radix 30 g, Zingiberis Rhizoma 10 g, Descurainiae Semen 15 g), one dose daily for consecutive 30 days. Before and after treatment the changes in levels of B-type natriuretic peptide (BNP), serum creatinine (SCr), blood urea nitrogen (BUN), amount of urine, clinical efficacy and TCM syndrome score efficacy were observed in two groups.Results After treatment, the levels of BNP, SCr, and BUN were significantly decreased, while urine volume was obviously increased compared with those before treatment in the two groups, and the degrees of changes in the treatment group were superior to those in control group [BNP (ng/L): 297.3±75.1 vs. 344.2±56.3, SCr (μmol/L): 139.7±62.1 vs. 154.4±39.7, BUN (mmol/L): 10.1±6.4 vs. 13.2±8.7, urine volume (mL/d): 847.2±32.7 vs. 786.4±13.6, allP < 0.05]. The total effective rates of patients and TCM syndrome scores in treatment group were significantly higher than those in control group [both 86.7% (13/15) vs. 66.7% (10/15), bothP < 0.05]. Conclusions Modified Linggui Zhugan decoction can alleviate the symptoms of yang deficiency of heart and kidney and heart failure due to the attack of heart by retained fluid, and can also ameliorate the complicated renal function impairment; the therapeutic effect of integrative traditional Chinese and western medicine for treatment of patients with type Ⅱ CRS is superior to that of conventional western medicine treatment.
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Objective: To study the mechanism of Linggui Zhugan Decoction (LZD) on chronic heart failure (CHF), through observing the expression of TNF-α and TNF-α mRNA in myocardial tissue and the levels of NF-κB and IL-1β in serum of model rats with CHF. Methods: Model of CHF was produced by ligation of coronary artery. Four weeks after this procedure, rats were randomly classified into model, Captopril (4.375 mg/kg), and low-, mid-, and high-dose (2.1, 4.2, and 8.4 g/kg) of LZD groups. Sham group and the other five groups were ig administered once daily for consecutive 4 weeks. The expression of TNF-α and TNF-α mRNA in myocardial tissue was detected by Western blotting and RT-PCR and the levels of NF-κB and IL-1β in serum were detected by ELISA. Results: The expression of TNF-α and TNF-α mRNA of myocardial tissue was enhanced and the levels of NF-κB and IL-1β in serum of model group were significantly increased compared with Sham group (P < 0.01). The low-, mid-, and high-dose of LZD and Captopril groups could inhibit the expression of TNF-α and TNF-α mRNA in myocardial tissue and decrease the levels of NF-κB and IL-1β in serum significantly compared with the model group (P < 0.05, 0.01). Conclusion: The mechanism of LZD interfering CHF seems to be related to the regulation of the cytokine network inflammatory factors.