Exploration of Spectrum-effect Relationship of Zhuriheng Dropping Pills Against Macrophage Foaming Based on UPLC-Q-Exactive Orbitrap MS / 中国实验方剂学杂志

ObjectiveThrough the correlation analysis between intestinal absorption profile and inhibition of macrophage foaming， the pharmacodynamic components of Zhuriheng dripping pills（ZRH） were explored to provide a basis for establishing its quality standard. MethodIntestinal absorption fluids with 0， 5， 10， 15， 20 times clinical equivalent doses were prepared by a rat everted gut sac（EGS）， and the oxidized low density lipoprotein（ox-LDL）-induced RAW264.7 macrophage foaming model was used to investigate the effect of intestinal absorption fluid with different doses on the accumulation of lipids in RAW264.7 cells by oil red O staining and cholesterol content determination， and to screen for the optimal dose. Ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to analyze and identify intestinal absorption fractions of ZRH intestinal absorption fluids， and partial least squares-discriminant analysis（PLS-DA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were performed on different doses of ZRH intestinal absorption fluids using SIMCA 13.0 with peak area as the independent variable and the pharmacodynamic indicators as the dependent variables to screen the compounds with variable importance in the projection（VIP） value>1.0 as contributing components， and Pearson correlation analysis was used to determine the spectral effect relationship， determined the compounds and positive correlation with pharmacodynamic were as active ingredients. Molecular docking was used to verify the binding energy of peroxisome proliferator-activated receptor α（PPARα）， PPARγ， PPARβ， human retinoid X receptor α（RXRA） and nuclear transcription factor-κB（NF-κB） with the active ingredients in ZRH intestinal absorption fluids. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was performed to detect the mRNA levels of PPARγ， scavenger receptor A1（SRA1） and adenosine triphosphate-binding cassette transporter A1（ABCA1） in RAW264.7 cells， Westen blot was used to detect the expression level of PPARγ protein in RAW264.7 cells， and enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of interleukin（IL）-1β and NF-κB in RAW264.7 cells. ResultAccording to the results of oil red O staining and cholesterol content determination， the ZRH intestinal absorption fluids could significantly reduce macrophage foaming， and intestinal absorption fluids with 15， 20 times clinical equivalent doses had the best effect， the 15-fold ZRH intestinal absorption fluid was finally determined as the study subject. Spectral effect relationship showed that 52 corresponding peaks in the ZRH-containing intestinal fluid were positively correlated with the efficacy， including organic acids， phenylpropanoids， iridoids， flavonoids， bile acids， coumarins and chromones. Target validation results showed that 86.9%-96.2% of the total components processed good binding activities with the key targets of PPARα， PPARγ， PPARβ， RXRA and NF-κB， and the docking energy values were all less than -6.0 kcal·mol-1（1 cal≈4.19 J）. The results of validation showed that， compared with the normal group， the model group showed a significant increase in the levels of SRA1 and PPARγ mRNA expression， a significant decrease in ABCA1 mRNA expression， a significant increase in the level of PPARγ protein expression， and a significant increase in the levels of IL-1β and NF-κB（P<0.01）， compared with the model group， the 15-fold intestinal absorption fluid group showed a significant decrease in the levels of SRA1 and PPARγ mRNA expression（P<0.05， P<0.01）， ABCA1 mRNA expression level was significantly up-regulated， the levels of IL-1β and NF-κB were significantly reduced（P<0.01）， and PPARγ protein expression level was significantly reduced（P<0.05）. ConclusionThis study identifies 52 components and their metabolites in ZRH intestinal absorption fluid that are positively correlated with the inhibition of macrophage foaming， which may be related to the regulation of the PPARs pathway in cells and the reduction of the levels of inflammatory factors， and can provide a reference for the quality control and clinical application of ZRH.

Protective effects of catalpol on cardio-cerebrovascular diseases:A comprehensive review / 药物分析学报

Catalpol,an iridoid glucoside isolated from Rehmannia glutinosa,has gained attention due to its potential use in treating cardio-cerebrovascular diseases(CVDs).This extensive review delves into recent studies on catalpol's protective properties in relation to various CVDs,such as atherosclerosis,myocardial ischemia,infarction,cardiac hypertrophy,and heart failure.The review also explores the compound's anti-oxidant,anti-inflammatory,and anti-apoptotic characteristics,emphasizing the role of vital signaling pathways,including PGC-1a/TERT,PI3K/Akt,AMPK,Nrf2/HO-1,estrogen receptor(ER),Nox4/NF-κB,and GRP78/PERK.The article discusses emerging findings on catalpol's ability to alleviate diabetic cardiovascular complications,thrombosis,and other cardiovascular-related conditions.Although clinical studies specifically addressing catalpol's impact on CVDs are scarce,the compound's established safety and well-tolerated nature suggest that it could be a valuable treatment alternative for CVD patients.Further investigation into catalpol and related iridoid derivatives may unveil new opportunities for devising natural and efficacious CVD therapies.

Research Progress on Anti-atherosclerosis Mechanism of Mongolian Medicine / 中国实验方剂学杂志

Atherosclerosis is one of the most common diseases that threaten human health. How to effectively inhibit atherosclerosis， extend the survival time and improve the quality of life has become one of the most urgent issues to be solved clinically. Mongolian medicine， with a long history of managing human diseases， is an important part in traditional Chinese medicine （TCM） and has distinct ethnic characteristics. It has been gradually formed and developed by absorbing some theories of Tibetan medicine， Indian medicine and relevant knowledge of TCM. Mongolian medicine has many advantages， including but not limited to， low toxicity and diverse structure. However， the action mechanism of Mongolian medicine in preventing and managing atherosclerosis has yet to be fully clarified， which has been a major obstacle for further promotion and application of Mongolian medicine in clinical settings. In this review， the up-to-date research findings on Mongolian medicine were collected， analyzed and summarized， and the anti-atherogenic action mechanism of Mongolian medicine were reviewed from the aspects of anti-inflammatory， lipid-lowering， anti-oxidative stress， vascular endothelial cell protection， and inhibition of vascular smooth muscle cell proliferation and migration.

Research progress on anti-atherosclerosis effect and mechanism of flavonoids compounds mediated by macrophages / 中国中药杂志

Flavonoids are important active ingredients of traditional Chinese medicine, mainly with cardiovascular, anti-liver injury, antioxidant, antispasmodic, and estrogen-like effects. These compounds have obvious effects on the cardiovascular and cerebrovascular diseases. Macrophage-derived foam cells are the key medium in the process of atherosclerosis(AS). In plaque, allserum lipids, serum lipoproteins, and various pro-or anti-inflammatory stimulating factors, chemokines, and small bioactive molecules can significantly affect the macrophage phenotype and induce stronger pro-inflammatory or anti-inflammatory properties. Studies have shown that some flavonoids can be used for macrophages through different pathways and mechanisms, playing an anti-atherosclerosis effect to different degrees, including promotion of cholesterol efflux from macrophages, anti-foaming of macrophages, inhibition of secretion of inflammatory factors, and antioxidant modified low density lipoprotein(ox-LDL)-induced apoptosis of macrophages. Related gene regulation inclu-ded ATP-binding cassette transporter A1(ABCA1), ATP-binding cassette transporter G1(ABCG1), Toll-like receptor(TLR), and scavenger receptor(SR). In this article, we would review the recent research progress of flavonoids on anti-atherosclerosis effect me-diated by macrophage. It is expected to provide new treatment strategies for AS-related cardiovascular and cerebrovascular diseases, and provide research ideas and development directions for the use of related natural medicines and design of new products.

Optimization of Eehanol Extraction Technology of Anti-atherosclerosis Effective Fraction in Lindley eupatorium by Multiple Index Scoring Method / 中国药房

OBJECTIVE:To optimize the ethanol extraction process of anti-atherosclerosis effective fraction of Lindley eupatorium. METHODS:Using the contents of hyperin,quercetin,kaempferol and jaceosidin as indexes,the effects of ethanol volume fraction,liquid-solid ratio,extraction time and extraction times on ethanol extraction technology were investigated. By multiple index scoring method,orthogonal test was designed to optimize extraction technology,and validation test was conducted. RESULTS:The optimal extraction technology was 70% ethanol as extraction solvent with solid-liquid ratio of 12 : 1,reflux extracting for 3 times,2 h each time. Results of 3 batches of validation tests showed that the contents of 4 marker components were in high level,being 5.305 3,1.560 1,1.986 5,6.703 6 mg/g in average(RSD<1.1%,n=3). Results of validation tests were stable and reliable,and were in good agreement with the results of orthogonal tests. CONCLUSIONS:The optimized ethanol reflux extraction technology of L. eupatorium is stable and reliable,and performs well. Energy consumption and cost are suitable for industrial production.The results provide technology basis for further purification and enrichment of anti-atherosclerosis effective components of L.eupatorium.

Protective effects of Xiongshao Capsule () on anti-inflammatory function of high-density lipoprotein in an atherosclerosis rabbit model / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Xiongshao Capsule (, XSC) on anti-inflflammatory properties of high-density lipoprotein (HDL), myeloperoxidase (MPO) and paraoxonase 1 (PON1) in serum of atherosclerosis (AS) rabbit model and explore the anti-inflflammatory protective effects of XSC on HDL.</p><p><b>METHODS</b>Sixty rabbits were randomized into the control, the model, XSC low-, medium- and high-dose (Rhizoma Chuanxiong + Radix Paeoniae rubra: 0.6+0.3, 1.2+0.6, 2.4+1.2g·kg·day, respectively), and simvastatin (1g·kg·day) groups. The model rabbits were fed with high-fat diet and drugs for 15 weeks. The blood and thoracic aortas samples were collected at the end of 15 weeks. The levels of serum MPO and PON1 as well as total cholesterol (TC) and free cholesterol (FC) in aorta wall cells were tested by enzyme linked immunosorbent assay.</p><p><b>RESULTS</b>TC and FC in the model group were significantly higher than those in the control group (P<0.01). Compared with the model group, TC and FC in the XSC groups were signifificantly lower (P<0.05 or P<0.01), so was simvastatin group (P<0.01). There was no signifificant difference in PON1 level between groups (P>0.05), even between model and control groups (P>0.05). The serum MPO level in the model group was signifificantly higher than that in the control group (P<0.05), which was signifificantly lower in XSC groups as well as simvastatin group (P<0.05 or P<0.01), and no difference was found between XSC groups and simvastatin group (P>0.05).</p><p><b>CONCLUSIONS</b>XSC can reduce the serum MPO level in AS rabbits to protect the anti-inflammatory function of HDL, maintaining the normal lipid transport function. TC and FC levels in aorta cells decline, and this process initiated by XSC plays an anti-AS role.</p>

Mechanism of anti-atherosclerosis of astragaloside IV based on regulation of macrophage autophagy by PI3K/Akt/mTOR signaling pathway / 中草药

Objective To study the mechanism of anti-atherosclerosis of astragaloside IV by observing the regulation of PI3K/Akt/mTOR signaling pathway. Methods In the in vitro experiments, macrophages were randomly divided into control group, astragaloside IV group, Kangshide group, rapamycin group and siRNA group. The changes of autophagy of macrophages were observed by transmission electron microscopy (TEM) after 48 h of mouse RAW264.7 macrophages in vitro treated by astragaloside IV-containing serum, Akt inhibitor Kangshide, mTOR inhibitor rapamycin and mTOR-siRNA. The expression of Akt, mTOR and autophagy-associated protein Beclin 1 was detected by real-time quantitative RT-PCR and Western blotting. The expression of Beclin 1 was detected by immunofluorescence and Western blotting. IL-4, IL-10, IL-2, and IFN-γ in RAW264.7 cells were detected by ELISA. In the in vivo experiments, the pathological changes of aorta were detected by HE staining. The expression of Akt, mTOR mRNA and protein in aorta of mice was detected by qRT-PCR and Western blotting, respectively. Results In vitro experimental results showed that compared with control group, the autophagus of astragaloside IV-containing serum group and each inhibitor group was significantly increased under the TEM (P < 0.05). The expression levels of LC3-II and Beclin1 were significantly up-regulated (P < 0.05). The expression of Akt and mTOR mRNA and protein was significantly decreased (P < 0.05). The secretion of IL-10 was significantly decreased and the secretion of IFN-γ was significantly increased (P < 0.05). HE staining results showed that, compared with the model group, the blood vessels of astragalus membranaceus group were normal, arranged neatly, with small focal calcification of the granules. The lesions were mild, the patches were small, the foam cells and the lipids were reduced, the elastic plates were basically complete, The degree of lesion was significantly lighter and lighter than that of the inhibitor groups. The expression of Akt, mTOR mRNA and protein were significantly lower in the aorta of astragaloside IV group (P < 0.05). Conclusion Astragaloside IV inhibition of atherosclerotic plaque formation mechanism and regulation of PI3K/Akt/mTOR signaling pathway, inhibition of inflammatory response.

Research progress in anti-cardiovascular and cerebrovascular disease activity of Citri Reticulatae Pericarpium / 中草药

Citri Reticulatae Pericarpium is a common Chinese materia medica, with extensive distribution, abundant resource, and high medicinal value. Epidemiologic studies showed that Citrus flavonoid intake can relieve the symptom of cardiovascular and cerebrovascular disease. Numerous in vitro and in vivo studies have found that Citri Reticulatae Pericarpium and its composition were good to treat cardiovascular and cerebrovascular disease, including hypolipidemic, anti-fatty liver, anti-thrombusis, anti-atherosclerosis, cardio-cerebral-vascular protection, and anti-oxidant and anti-inflammatory effects closely related to the above. Evidence obtained from these studies indicates that Citri Reticulatae Pericarpium has the potential in the prevention and treatment of cardiovascular and cerebrovascular disease.

Research progress on hypolipidemic constituents in Alismatis Rhizoma and their pharmacology and clinical application / 中草药

Alismatis Rhizoma, as a Chinese materia medica with multiple uses, has a long history of application in China. By retrieving the published literature of hypolipidemic effect, this review presents an overview of Alismatis Rhizoma, including chemistry, pharmacological mechanisms, clinical application, and safety evaluation. The protostane triterpenoids, as alisol A and alisol A 24-acetate, had hypolipidemic effects. Early preliminary clinical studies have shown that the Alismatis Rhizoma extraction may be a good hypolipidemic drug. Modern pharmacologic study found that it had anti-atherosclerotic effects. The reported safety evaluation results showed that the extraction was with low liver and kidney toxicity and might be suitable for long term use. We think it is worth to be developed a new drug.

Research progress on chemical constituents of plants from Mallotus Lour. and their pharmacological activities / 中草药

The genus Mallotus Lour. (Euphorbiaceae) consists of 140 species distributed in tropical and subtropical regines of Asia, 25 species and 11 varieties of which can be found in southern of China. Some species of the genus have been used in traditional medicine. Phytochemical studies on the plants in the genus have demonstrated the presence of large number of diterpenoids, triterpenoids, polyphenols, and tannins, along with some monoterpene, benzopyran, coumarin-lignin, diaryl heptane, flavonoids, alkaloids, coumarin, etc. Furthermore, many pharmacological activities of the plants in the genus, such as anti-atherosclerosis, choleretic, hepatoprotective, antioxidant, analgesic, immunity improvement, and memory enhancing activities have been reported. In this article, new development of constituents and biological activities of plants in Mallotus Lour. since 1988 were reviewed and summarized for providing a reference for the further study, exploitation, and utilization of this resource.

Progress in the study of pharmacology and application of chondroitin sulfate / 中国药学杂志

OBJECTIVE: To review the pharmacologic action and application of chondroitin sulfate.