Inhibitory Effect of Paeonol on Vascular Smooth Muscle Cell Senescence by Reducing SIRT6/PARP1-mediated DNA Damage / 中国实验方剂学杂志

ObjectiveTo investigate whether the effects of paeonol （Pae） on angiotensin Ⅱ （AngⅡ）-induced senescence in vascular smooth muscle cells （VSMCs） were related to angiotensinogen of silencing regulatory information factor 6 （SIRT6）/adenosine diphosphate ribose polymerase 1 （PARP1） signaling pathway in VSMCs. MethodThe model of VSMC-stress aging induced by AngⅡ （100 nmol·L-1） was established. The rats were divided into normal group， model group， low， medium， and high-concentration Pae groups （30， 60， 120 μmol·L-1）. The positive rate of cell senescence was detected by SA-β-Gal staining， the ability of cell proliferation was detected by the cell counting kit-8 （CCK-8） method， the expression of SIRT6， PARP1， p16， p21， p53， proliferating cell nuclear antigen （PCNA）， deoxyribonucleic acid （DNA）-damaged protein γ-H2AX was detected by Western blot， and VSMC proliferation was detected by EdU staining. The silenced VSMCs were prepared by siRNA-SIRT6 transfection， and the protein expressions of SIRT6， PARP1， p16， and γ-H2AX in VSMCs silenced by SIRT6 were observed. ResultThe results of SA-β-Gal staining showed that the senescence positive rate of SA-β-Gal staining in the model group was higher than that in the normal group （P<0.01）， and the positive rate of SA-β-Gal staining in the Pae group was significantly lower than that in the model group （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the expression of PCNA， SIRT6， and PARP1 in the model group was down-regulated， and the expression of aging-related proteins p16， p21， p53， and γ-H2AX was up-regulated in the model group （P<0.05， P<0.01）. Compared with the model group， Pae promoted the protein expression of PCNA， SIRT6， and PARP1 and inhibited the protein expression of p16， p21， p53， and γ-H2AX in a dose-dependent manner （P<0.05， P<0.01）. The results of EdU staining showed that the number of EdU positive cells in the model group was lower than that in the normal group （P<0.01）， and the number of EdU positive cells in Pae groups was significantly higher than that in the model group （P<0.05， P<0.01）. After SIRT6 silencing， the effects of Pae on promoting SIRT6 and PARP1 and inhibiting P16 were reversed （P<0.05， P<0.01）. In addition， the addition of SIRT6 inhibitor （IN-1） promoted the occurrence of cell senescence induced by AngⅡ （P<0.05， P<0.01）. ConclusionPae can effectively inhibit the aging of VSMCs， and its mechanism may be related to the regulation of SIRT6/PARP1 signal pathway.

Preparation of paeonol nanoemulsion and investigation of vascular endothelial cells uptake / 中国药科大学学报

@#In order to improve the poor solubility and low bioavailability of paeonol (Pae), paeonol-nanoemulsion (Pae-NE) was prepared, and its effect on uptake of human umbilical vein endothelial cells (HUVECs) was investigated.Pae-NE was prepared by phase inversion composition (PIC), the formulation of Pae-NE was optimized by single factor method and central composite design-response surface method (CCD), and the pharmaceutical properties were further characterized.Moreover, MTT was applied to evaluate the toxicity of Pae-NE on HUVECs, and the cellular uptake efficiency of Pae-NE was detected by fluorescence microscopy and flow cytometry.The results showed that the optimal formulation of Pae-NE was 20 mg of Pae, 55.1 mg of LCT, 144.9 mg of MCT, 600 mg of HS15, and 200 mg of 1,2 propylene glycol.The Pae-NE appearance was a light blue emulsion, and the average particle size is (25.69 ± 0.03) nm, with PDI of 0.182 ± 0.09, Zeta potential of -(4.01 ± 0.30) mV and good stability.The drug loading of Pae-NE was (1.967 ± 0.28) mg/mL and encapsulation rate of (99.36 ± 0.1)%.Pae-NE performed no significant effect on HUVECs growth in the Pae concentration range of 10-1-10-3 μg/mL.Moreover, NE as a drug delivery carrier significantly enhanced the uptake efficiency of Pae on HUVECs.In conclusion, Pae-NE preparation method was simple and stable, and promotes HUVECs uptake efficiency of Pae, suggesting that NE was a better dosage form reference for the lipid-soluble drug of Pae.

Comparative study on in vivo and in vitro permeability of Huoxue Zhitong gel patch and microemulsion gel / 中国中药杂志

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.

Effects of components in stasis-resolving and collateral-dredging Chinese herbal medicines on angiogenesis and inflammatory response of human umbilical vein endothelial cells induced by VEGF / 中国中药杂志

The present study investigated the mechanism of components in stasis-resolving and collateral-dredging Chinese herbal medicines, including scutellarin(Scu), paeonol(Pae), and hydroxy safflower yellow A(HSYA), in the treatment of psoriasis by regulating angiogenesis and inflammation. The human umbilical vein endothelial cells(HUVECs) cultured in vitro were divided into a normal group, a model group, a VEGFR tyrosine kinase inhibitor Ⅱ(VRI) group, and Scu, Pae, and HSYA groups with low, me-dium, and high doses. Cell viability was detected by the CCK-8 assay. Cell migration was detected by wound healing assay. Tube formation assay was used to measure the tube formation ability. Western blot was used to detect the protein expression of the VEGFR2/Akt/ERK1/2 signaling pathway. The secretion levels of inflammatory cytokines IFN-γ, IL-1β, IL-6, and TNF-α were detected by ELISA. The results showed that compared with the model group, all the Scu, Pae, and HSYA groups could reduce cell viability, inhibit cell migration and tube formation(P<0.05, P<0.01), and down-regulated the protein expression of VEGFR2, p-VEGFR2, Akt, p-Akt, ERK1/2, and p-ERK1/2. Scu and Pae could down-regulate VEGFR2 expression(P<0.05, P<0.01), while other groups only showed a downward trend. Scu and Pae significantly reduced IFN-γ and IL-6 levels(P<0.01), and HSYA significantly reduced the levels of IFN-γ, IL-1β, and IL-6(P<0.01). Scu, Pae, and HSYA had no significant effect on TNF-α. The results suggested that Scu, Pae, and HSYA may exert a therapeutic role in psoriasis-related angiogenesis and inflammation by inhibiting VEGFR2/Akt/ERK1/2 signaling pathway and inhibiting the secretion of IFN-γ, IL-1β, and IL-6.

HPLC fingerprint of Moutan Cortex in different growth period / 国际中医中药杂志

Objective:To establish the HPLC fingerprint method for assessing the quality of Moutan Cortex, and to determine the contents of paeonol, paeoniflorin, gallic acid, hydroxyl-paeoniflorin and benzoyl-paeoniflorin of Moutan Cortex in different growth period. Methods:Diamonsil Plus C18 column (250 mm × 4.6 mm, 5 μm) was used with the mobile phase comprising acetonitrile-0.05% formic acid solution and the flow rate of 1.0 ml/min with gradient elution manner. The detected wavelength was 230 nm for paeoniflorin and benzoyl-paeoniflorin, 267 nm for gallic acid, 258 nm for hydroxyl-paeoniflorin and 274 nm for paeonol with temperature column of 25 ℃. Then putting chromatograms into Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica (2012A) to evaluate the similarity of Moutan Cortex in different growth period; then putting peak area data into SPSS software for cluster analysis and the clustering effect was determined. Results:The HPLC fingerprints established with this method has 23 shared peaks and 5 of them were identified, namely, paeonol, paeoniflorin, gallic acid, hydroxyl-paeoniflorin and benzoylpaeoniflorin. The similarity of Moutan Cortex in different years was between 0.850-0.991. This method has good linear relation ( r≥0.999 5), RSDs of precision, stability tests and reproducibility were lower than 1.6% ( n=6). Different growth periods of Moutan Cortex have obvious influence on the concentration of five compounds. Conclusion:This method is useful to evaluate and discriminate Moutan Cortex at different growth periods so as toprovide scientific reference on the harvest,industrialization and evaluation of Moutan Cortex.

Discussion for Quality Control of Chinese Patent Drugs from Quality Difference Among Different Dosage Forms of Liuwei Dihuang Preparations / 中国实验方剂学杂志

Objective:Through comprehensive evaluation and analysis of the quality of Liuwei Dihuang （LWDH） preparations from different manufacturers and combining factors such as production technology， the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations. Method:Morroniside， loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A） -0.3% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-8%A； 5-20 min， 8%A； 20-35 min， 8%-20%A； 35-45 min， 20%-60%A； 45-55 min， 60%A）， the detection wavelength of paeonol was at 274 nm， and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms （big candied pills， water-honeyed pills， concentrated pills， hard capsules and soft capsules） from different manufacturers were analyzed. Combined these results with their actual production processes， the key-points of quality control in the whole production process were discussed. Result:The contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of <italic>Chinese Pharmacopoeia</italic>， however， the content limit of some dosage forms in the current standard was unreasonable. For example， although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills （15.00， 8.57 g， respectively）， they got exactly the same daily limits of the contents for both the quality markers. What′s more， these two formulations had the same process， so the differences between the process obviously could not be the reason of these differences. Conclusion:It is recommended that for the products with different dosage forms should have a similar content limits， if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards， and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.

Mechanism of paeonol combined with paeoniflorin against myocardial ischemia injury:based on proteomics / 中国中药杂志

The study aims to investigate the effect of the compatibility of paeonol and paeoniflorin(hereinafter referred to as the compatibility) on the expression of myocardial proteins in rats with myocardial ischemia injury and explore the underlying mechanism of the compatibility against myocardial ischemia injury. First, the acute myocardial infarction rat model was established by ligation of the anterior descending branch of the left coronary artery. The model rats were given(ig) paeonol and paeoniflorin. Then protein samples were collected from rat cardiac tissue and quantified by tandem mass tags(TMT) to explore the differential proteins after drug intervention. The experimental results showed that differential proteins mainly involved phagocytosis engulfment, extracellular space, and antigen binding, as well as Kyoto encyclopedia of genes and genomes(KEGG) pathways of complement and coagulation cascades, syste-mic lupus erythematosus, and ribosome. In this study, the target proteins and related signaling pathways identified by differential proteomics may be the biological basis of the compatibility against myocardial ischemia injury in rats.

Effects of Paeonol Combined with Cisplatin on the Proliferation and Apoptosis of Human Osteosarcoma MG- 63 Cells as Well as PI 3K/Akt/mTOR Signaling Pathway / 中国药房

OBJECTIVE：To study the effects of paeonol combined with cisplatin on the proliferation and apoptosis of human osteosarcoma cell MG- 63 and its possible mechanism. METHODS ：MG-63 cells in logarithmic growth phase were divided into blank control group ，cisplatin group （4 µmol/L），paeonol group （50 mg/L），and low ，medium，high concentration combined groups （50，100，200 mg/L paeonol+ 4 µmol/L cisplatin ）. CCK- 8 method was used to detect the cell proliferation rate at 24，48，and 72 hours of treatment. Annexin Ⅴ-FITC/PI double staining method was used to detect the cell apoptosis rate at 24 hours of treatment. The relative expression of PI 3KCA，Akt，mTOR，P-gp and PTEN mRNA in cells were detected by qRT-PCR. RESULTS ：Compared with blank control group ，the cell proliferation rate at each time point ，and the relative expression of PI 3KCA，Akt and mTOR mRNA in cells were significantly reduced and the apoptosis rate and the relative expression of P-gp and PTEN mRNA in the cells were increased significantly （P＜0.05 or P＜0.01）. Compared with cisplatin group and paeonol group ，cell proliferation rate at each time point and the relative expression of PI 3KCA，Akt and mTOR mRNA in cells were decreased significantly in the high concentration combination group ，while the relative expression of P-gp and PTEN mRNA in the cells were significantly increased （P＜0.01）；there were statistical significance in some of the above indicators in the medium and low concentration combination groups（P＜0.05 or P＜0.01）. CONCLUSIONS ：The combination of paeonol and cisplatin could inhibit the proliferation of MG- 63 cells and promote their apoptosis ，which may be related to the down-regulation of PI 3KCA，Akt and mTOR mRNA expression ，and the up-regulation of P-gp and PTEN mRNA expression.

Paeonol attenuated vascular fibrosis through regulating Treg/Th17 balance in gut microbiota-dependent manner / 中国药理学与毒理学杂志

OBJECTIVE Atherosclerosis (AS) is a chronic inflammatory disease characterized by the accumulation of lipids, vascular fibrosis, and inflammation. Paeonol (Pae) is a natural phenolic compounds isolated from a traditional Chinese medicine, Cortex Moutan, which exhibits anti-AS effects. Our previous work demonstrated that gut microbiota plays an important role during AS treatment as it affects the efficacy of Pae. However, the mechanism of Pae in protect?ing against vascular fibrosis as related to gut microbiota has yet to be elucidated. To investigate the anti-fibrosis effect of Pae on AS mice and demonstrate the underlying gut microbiota-dependent mechanism. METHODS ApoE-/- mice were fed with high-fat-diet (HFD) to replicate the AS model. HE and Masson staining were used to observe the plaque forma?tion and collagen deposition. Gut microbiota alteration and short-chain fatty acids (SCFAs) production were analyzed through 16S rRNA sequencing and LC-MS/MS. The frequency of immune cells in spleen were phenotyped by flow cytometry. The mRNA expression of aortic inflammatory cytokines were detected by qRT-PCR. The protein expression of LOX and fibrosis related indicators were examined by Western blotting. RESULTS Pae restricted the development of AS and collagen deposition. Notably, the anti-fibrosis effect of Pae was achieved by regulating the gut microbiota. 16S rRNA sequencing and LC-MS/MS data indicated that the relative abundance of SCFAs-producing bacteria and SCFAs production was increased. Additionally, Pae administration selectively up-regulated the frequency of regulatory T (Treg) cells as well as down-regulated the ratio of T helper type 17 (Th17) cells in the spleen of AS mice, improving the Treg/Th17 balance. In addition, as expected, Pae intervention significantly down-regulate the mRNA expression levels of pro-inflammatory cytokines IL-1β, IL-6, TNF-αand IL-17 in the aorta tissue, up-regulate the levels of anti-inflammatory factor IL-10, a marker of Treg cells. Finally, Pae's intervention in the gut microbiota resulted in the restoration of the balance of Treg/Th17, which indirectly down-regulated the protein expression level of LOX and fibrosis-related indicators (MMP-2/9 and collagenⅠ/Ⅲ). CONCLUSION Pae attenuates vascular fibrosis in a gut microbiota-dependent manner. The under?lying protective mechanism is associated with the improved Treg/Th17 balance in spleen mediated through the increased microbiota-derived SCFAs production.

On-line moisture determination and optimization of drying process of Qiju Dihuang Pills by time-domain reflection method / 中草药

Objective: Taking Qiju Dihuang Pills (QDP) as the research object, time domain reflection method was used for real-time determination of moisture content in concentrated pills during drying process and optimization of the drying process parameters. Methods: The moisture model of the drying process of QDP was established by the relationship between the water, temperature, and the reflective signal value of time domain reflector. The effect of the drying process on the different thickness (8, 16, and 24 mm), different drying temperatures (30, 40, 50, 60, 70, 80, and 90℃) was investigated. Results: The moisture model of the drying process of QDP was measured by time domain reflection method as Y = 0.305 X-34.772 (r2 = 0.999); X = X(T)-(0.768 9 T-24.824 7) (T ≥ 30℃). The optimized process was as following: the process was dried at 60℃ to 13.8% moisture and then rising to 80℃, after being dried to 7.80%, cooled to 60℃ and dried to 5.0% target moisture. Conclusion: It is feasible to test the moisture content in the drying process of QDP by time domain reflection method. This method can be used to monitor and popularize the moisture content in the drying process of traditional Chinese medicine concentrated pills.

Optimization of formulation of paeonol lipid microspheres by central composite design-response surface method and its drug release mechanism in vitro / 中草药

Objective: To optimize the formulation of paeonol lipid microspheres (Pae-LM) through central composite design- response surface method and determine its in vitro release characteristics. Methods: Using the mean particle size and centrifugal stability constant (Ke) as evaluation indexes, the oil phase type and the ratio of composite oil, the amount of phospholipid and stearic acid, the type of emulsifier, the type and amount of stabilizer, the quality of PC and CH, the high-speed shear temperature and time, the homogenization pressure and time was screened in prescription process. Effects of dosage of paeonol and high pressure homogenizing pressure on the properties of Pae-LM preparation were investigated by central composite design-response surface method. The binomial model and multivariate linear regression model were used to establish the mathematical relationship between the indexes and the factors. According to the best mathematical model of evaluation index, the response surface was depicted and the best prescription was analyzed by the response surface method. According to the optimized formulation Pae-LM, the in vitro drug release characteristics were investigated. Results: The best prescription of Pae-LM was basically round, with mean particle size of (149.32 ± 0.57) nm, Zeta potential of (-36.01 ± 3.09) mV, encapsulation rate of (98.24 ± 0.32)% and drug-loading rate of (11.94 ± 0.04)%. There was a credible quantitative relationship between Ke and the two factors, and the binomialmodel was more reliable than the multivariate linear model. The cumulative release of paeonol drug substance at 12, 24 and 36 h were 71.84%, 85.21% and 95.07%, while the cumulative release of Pae-LM was only 57.21%, 59.66%, and 63.91% at 12, 24 and 36 h, respectively. The drug release was in accordance with the Ritger-peppas model. Conclusion: Central composite design-response surface method can be applied to optimize prescription of lipid emulsion microspheres. The optimized particle size of Pae-LM was suitable with a higher encapsulation rate, which can provide a reference for the development of paeonol cardiovascular delivery system.

Study on specific chromatogram and content determination of multi-index components about material reference of classic prescription of Huaganjian / 中草药

Objective: To establish the UPLC specific chromatogram and HPLC content determination methods of multi-index components about the material reference of classical Huaganjian and build its quality control system. Methods: According to the ancient books and combining with the previously inspected process, 18 batches of Huaganjian material reference from different origins were prepared. The specific chromatogram was established by using UPLC. Similarity was calculated by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2012). Combining with orthogonal partial least squares discriminant analysis, we excavated the main components that affected the quality of Huaganjian material reference from different batches and origins. Three of these index components (paeoniflorin, hesperidin, paeonol) from prescription sovereign drug, minister drug, and assistant drug were selected and used as indicators for content determination of Huaganjian material reference. HPLC content determination methods were established and the content of 18 batches of samples was determined respectively. Results: The similarity of the specific chromatogram was ≥ 0.989. Thirty-three common peaks were calibrated, and eight common peaks were identified by chemical composition (gallic acid, geniposide, paeoniflorin, hesperidin, didymin, paeonol, sinensetin, and 3,5,6,7,8,3',4'- heptamethoxyflavone). Nine index components that affected the stability between batches were found out (Peak 31, 20, 11, 13, 22, 33, 21, 29, 1). Paeoniflorin, hesperidin, and paeonol were selected as content determination indicators. The content range of these components in material reference was 1.28%-1.95% paeoniflorin, 0.91%-1.02% hesperidin, 0.48%-0.57% paeonol. Conclusion: The quality control method of the material reference of classic prescription Huaganjian was established preliminarily through the UPLC specific chromatogram and HPLC content determination of index components. This method was rapid, simple, feasible, reproducible, stable and could provide a theoretical basis for the subsequent development and quality control of Huaganjian preparations.

The parametric measurementof physico-chemical parameter and three types of effective components in Jiedu-Zhixue Decoction / 国际中医中药杂志

Objective@#To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters.@*Methods@#By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used.@*Results@#The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889.@*Conclusions@#The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.

Inhibitory effect of paeonol on aortic endothelial inflammation in atherosclerotic rats by up-regulation of caveolin-1 expression and suppression of NF-κB pathway / 中国中药杂志

To explore whether paeonol can play an anti-atherosclerotic role by regulating the expression of aortic caveolin-1 and affecting NF-κB pathway, so as to inhibit the inflammatory response of vascular endothelium in atherosclerotic rats. The atherosclerotic model of rats was induced by high-fat diet and vitamin D_2. The primary culture of vascular endothelial cells(VECs) was carried out by tissue block pre-digestion and adherent method. The injury model of VECs was induced by lipopolysaccharide(LPS), and filipin, a small concave protein inhibitor, was added for control. HE staining was used to observe pathological changes of aorta. TNF-α, IL-6 and VCAM-1 were detected by ELISA. Western blot assay was used to detect the protein expression levels of caveolin-1 and p65 in aorta and VECs. The results showed that as compared with model group, paeonol significantly reduced aortic plaque area and lesion degree in rats, decreased the level of serum TNF-α, IL-6 and VCAM-1 in the rats and enhanced the relative expression level of caveolin-1, decreased p65 expression conversely(P<0.05 or P<0.01). In vitro, as compared to model group, paeonol obviously improved cell morphology, decreased the secretion of TNF-α, IL-6 and VCAM-1 in VECs, increased caveolin-1 expression, and decreased p65 protein expression(P<0.05 or P<0.01). Furthermore, filipin could reverse the effect of paeonol on expression of inflammatory factors and proteins(P<0.05 or P<0.01). According to the results, it was found that paeonol could play the role of anti-atherosclerosis by up-regulating the expression of caveolin-1 and inhibiting the activation of NF-κB pathway to reduce vascular inflammation in atherosclerotic rats.

Paeonol inhibits macrophage M1 polarization by down-regulating miR-155/JAK1-STAT1 pathway / 中国中药杂志

The aim of this paper was to investigate the effect and mechanism of paeonol on peritoneal macrophage M1 polarization in mice, explore whether the intervention action is related to the down-regulation of miR-155 and the inhibition of downstream JAK1-STAT1 pathway, and provide a new idea for the molecular mechanism of paeonol against atherosclerosis(AS). Lipopolysaccharide(LPS) and interferon-γ(IFN-γ) were used to stimulate macrophages for 24 hours to establish the M1 polarization model, and paeonol was given 24 hours before co-stimulation to provide a pre-protective effect on cells. CCK-8 assay was used to detect the cells damage induced by LPS and IFN-γ co-stimulation; flow cytometry was used to detect the expression of M1 surface markers F4/80 and CD86. ELISA was used to detect the secretion of interleukin 6(IL-6) and tumor necrosis factor-α(TNF-α) in supernatant. RT-qPCR was used to detect the expression of miR-155, and Western blot was used to detect the protein expression at JAK1-STAT1-SOCS1 pathway. The results showed that LPS and IFN-γ had no obvious damage to the cells at the optimal concentration, but they induced macrophages polarized to M1, resulted in high expression of M1 type marker factors F4/80 and CD86 on the cell surface, and increased secretion of IL-6 and TNF-α on the cell surface(P<0.05 or P<0.01). Paeonol significantly reduced the LPS and IFN-γ-induced high expression of F4/80 and CD86, the secretion of inflammatory factors IL-6 and TNF-α(P<0.05 or P<0.01), decreased the expression level of miR-155, significantly down-regulated the protein phosphorylation level of JAK1-STAT1 and up-regulated the protein expression of SOCS1(P<0.01) in RAW264.7 cells. The results showed that paeonol could inhibit M1 polarization of macrophages by down-regulating cell surface marker factors and inflammatory factors secreted by cells, which may be related to the down-regulation of miR-155 expression and the inhibition JAK1-STAT1 pathway activation.

Effect of paeonol on adip/camkk(i/ampk pathway in alcoholic fatty liver in mice / 中国药理学通报

Aim To investigate whether the Adip/CaMKKp/AMPK pathway is involved in the treatment of alcoholic fatty liver in mice treated with paeonol. Methods After administration of paeonol on the basis of the mouse alcoholic fatty liver model, the inflammatory factors and TG and TC were detected by Elisa method to verify the validity of the model and drug treatment; meanwhile, the expression levels of Adip and ACC were detected by Elisa, and the nucleic acid and protein levels of Adip, CaMKK{3 and AMPKaby PCR and WB. Results Adip expression levels did not change significantly in each treatment group. Compared with control group, as for the expression of CaMKKp and AMPKa at the nucleic acid and protein-phosphorylation levels, ihe ElOH group was down-regulated to varying degrees, and the Sil and Pae groups improved the phenomenon to varying degrees. Compared with control group, as for the expression of SREBPlc at the nucleic acid and protein levels, the EtOH group was up-regulated to varying degrees, and the Sil and Pae groups improved the phenomenon to varying degrees. Conclusions The changes of theAdip/CaMKKp/AMPK signaling pathway confirm that pae-onol reduces steatosis injury of alcoholic fatty liver and inflammatory level in mice via this pathway.

Effect of paeonol on expression of HMGB1,IL-1βand apoptosis of cardiomyocytes in rats after myocardial I/R injury / 中华老年心脑血管病杂志

Objective To study the effect of paeonol onexpression of HMGB1and IL-1βand apoptosis of cardiomyocytes in rats after myocardial I/R injury.Methods Fifty-two adult SPF male SD rats were randomly divided into sham operation group,low-dose paeonol group,high dose paeonol group,and I/R injury group(13in each group).The rats in low-dose paeonol group and high dose paeonol group received intraperitoneal paeonol injection before operation(15.30mg/kg,once a day).Serum samples were taken at 2hafter reperfusion.Serum IL-1βlevel was measured by ELISA,myocardial infarction size was measured with TTC staining,expressions of HMGB1,Bcl-2and Bax proteins in myocardial tissue were detected with immunohistochemical staining.Results The myocardial infarction size was significantly smaller in low-dose paeonol group and high dose paeonol group than in I/R injury group(31.04％±2.93％,27.97％±2.84 vs37.23％±3.62％,P＜0.01).The expression levels of IL-1βand HMGB1were significantly higher in I/R injury group,low-dose paeonol group and high-dose paeonol group than in sham operation group and were significantly lower in low-dose paeonol group and high-dose paeonol group than in I/R injury group(P＜0.01).The expression level of IL-1βwas significantly higher while that of Bax protein was significantly lower in low-dose paeonol group and high-dose paeonol group than in in I/R injury group(P＜0.05).Conclusion Paeonol preconditioning can reduce apoptosis of cardiomyocytes,and can thus prevent myocardial I/R injury.

Chemical components and pharmacological effects for Guizhi Fuling Prescription and predictive analysis on quality marker (Q-marker) / 中草药

Guizhi Fuling Prescription is a classic prescription for promoting blood activation and resolving stasis. It has the functions of improving blood rheology, regulating endocrine, anti-inflammation, anti-cancer, and analgesia. In ancient times, it was used to treat women’s bleeding during pregnancy, abdominal pain, and postpartum lochia. It is also often used to treat gynecological diseases in modern clinics, such as uterine fibroids, ovarian cysts, dysmenorrhea, endometriosis and so on. This paper reviewed the research progress of chemical constituents, pharmacological activities, clinical application, and quality control of Guizhi Fuling Prescription in recent years. On the basis of this, the quality markers of Guizhi Fuling Prescription are predicted and analyzed based on the “five principles” of quality markers of Chinese materia medica in order to provide reference for the study of quality control of Guizhi Fuling Prescription.

Paeonol derivatives and their pharmacological activities / 中草药

Paeonol has a wide range of pharmacological activities. It is mainly used to protect cardiovascular and cerebrovascular system, promote microcirculation, antibacterial and anti-inflammatory, antitumor, anti-oxidation, anti-allergic reaction, enhance immunity, and treat diabetic osteoporosis, etc. In recent years, it not only improves the method of artificially synthesizing paeonol, but also carries out a large number of structural modification and transformation of paeonol. Five major parts, carbonyl, phenolic hydroxyl, carbonyl α-H, methoxy, and benzene rings, have been mainly modified. About 16 kinds of paeonol derivatives were produced including carboxylic acid esters, sulfonates esters, phosphates esters, ethers, NO donors, glycosidation, paeonol sodium, Schiff bases, thiazoles, oximes, hydroxyalkylation, α,β-unsaturated ketones, aminomethylation, α-H halogenation, demethylation, and benzene ring halogenation. In this paper, the derivatives of paeonol and their pharmacological activities were reviewed in combination with the literatures on paeonol derivatives at home and abroad. This article may provide basis for further study of this type of compound.

Optimization of extraction process of Modified Yangyin Qingfei Decoction by Box-Behnken response surface methodology based on analytic hierarchy process / 中草药

Objective: To optimize the extraction process of Modified Yangyin Qingfei Decoction (MYQD). Methods: The effects of ethanol concentration, extraction time and extraction times on the optimization of extraction process of MYQD were investigated by multiple indicators comprehensive scoring and Box-Behnken response surface methodology. The content of verbascoside, chlorogenic acid, paeonol, and glycyrrhizic acid was simultaneously determined by HPLC, and the method of analytic hierarchy process was used to calculate the weight coefficient. Results: The optimum extraction process was as follow: using 69% ethanol for once extraction for 68 min. Conclusion: The optimum extraction process is simple and feasible, and the extraction efficiency of components is high, which can provide reference for the extraction process of MYQD.