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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.
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OBJECTIVE To study the intervention effect and metabolic mechanism of Mongolian medicine Echinops sphaerocephalus extract on D-galactose-induced osteoporosis. METHODS Thirty-six 12-week-old male Wistar rats were selected and randomly divided into blank group， model group， Gushukang group， E. sphaerocephalus high-dose， medium-dose and low- dose groups， with 6 rats in each group. Except for blank group， other groups were intraperitoneally injected with D-galactose at 120 mg/kg per day. After 8 weeks of continuous injection， E. sphaerocephalus high-dose， medium-dose and low-dose groups were given drugs intragastrically at dose of 878， 439， 219.5 mg/kg， respectively. Gushukang group was given Gushukang 105.1 mg/kg intragastrically， once a day， for consecutive 8 weeks. After last administration， blood was collected from the abdominal aorta. Enzyme-linked immunosorbent assay was used to measure the contents of bone metabolism indexes [hydroxyproline （HYP）， alkaline phosphatase （ALP）] and oxidative stress indexes [total antioxidant capacity （TAOC）， superoxide dismutase （SOD）， malondialdehyde （MDA）] in serum of rats. Positron emission tomography/computedtomography （PET/CT） was used to analyze the changes of bone microstructure in right tibia bone. Meanwhile， metabolomic technology was used to study the regulation effect of E. sphaerocephalus on osteoporosis model rats. RESULTS Compared with blank group， HYP， ALP， MDA， ratio of bone surface to bone volume （BS/BV）， and trabecular separation （Tb·Sp） in model group were significantly increased （P＜0.05）， while TAOC， SOD， bone mineral density （BMD）， bone volume fraction （BVF）， trabecular E-mail：Xpfdc153@163.com thickness （Tb·Th） and trabecular number （Tb·N） were significantly decreased （P＜0.05）. Compared with model group， above indexes of administration groups were all reversed to different extents. The results of metabonomics study showed that after intervened with the extract of E. sphaerocephalus， 18 metabolites such as arachidonic acid， phenylalanine， tyrosine， tryptophan， isoleucine and uric acid in the serum of rats changed significantly， involving 15 metabolic pathways such as arachidonic acid， phenylalanine and tyrosine， of which arachidonic acid metabolism， phenylalanine metabolism and tyrosine metabolism were the main influencing pathways. CONCLUSIONS E. sphaerocephalus extract can effectively improve D-galactose-induced oxidative stress and the deterioration of bone microstructure， which interferes with metabolic pathways such as arachidonic acid metabolism and amino acid metabolism.
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Objective@#To study the effects of Phellodendron amurense on herpes simplex virus 1 (HSV-1) and cytokines, and to explore the mechanism of Phellodendron amurense inhibiting HSV-1 virus through multiple channels.@*Methods@#Viruses were inoculated into medicine treated HeLa cells. The proliferation of virus was observed by fluorescence microscopy. The transcriptional levels of glycoprotein gD and functional protein US1 on the surface of virus envelope were detected by quantitative (q) PCR. After incubating HeLa cells for 24 h, qPCR was used to detect the expression of intrinsic immune factors such as IP-10, IL-12, IFN-gamma and transcription factor NF-kappa B (P65). The expression and nuclear location of NF-kappa B (P65) protein were detected by immunofluorescence.@*Results@#Fluorescence showed that the proliferation of virus decreased significantly at 8 and 40 ng/ml (P<0.01), and the transcription levels of viral protein gD and US1 decreased (P<0.05). After incubation for 24 hours, the transcription levels of IP-10, IL-12 and IFN-gamma in HeLa cells increased significantly (P<0.01). The transcription level of transcription factor NF-kappa B (P65) also increased (P<0.05), and immunofluorescence showed that the nuclear penetration rate of p65 subunit of NF-kappa B (P65) in each group increased (P<0.05).@*Conclusions@#Phellodendron amurense extract can inhibit HSV-1 by inhibiting the transcription of viral functional protein and promoting the expression of cellular immune factors.
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@#Inflammatory cytokines are closely related to the development of gastric cancer. Pro-inflammatory cytokines, anti-inflammatory cytokines and their associated signaling pathways play different roles on the stages of gastric cancer, such as occurrence, development and prognosis, through different channels. This article summarizes the expression and function of Helicobacter pylori, IL-1β, TNF-α, IL-8, TGF-β, IL-10, IL-18 and T lymphocytes in gastric tissue or blood. The inflammatory cytokines and their associated signaling pathways which working as therapeutic targets for gastric cancer are also concluded.
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Objective To study the cardioprotective function of Zhurihen dripping pills in experimental animals.Methods By hypoxia tolerance model and isoproterenol hydrochloride induced acute hypoxia model, measuring the time of death in mice.For rat serum LDH, CK, SOD and MDA were measured to observe protective effect of Zhurihen dripping pills on vasopressin-induced acute myocardial ischemia model.The effect of Zhurihen dripping pills on latency and durations were observed in acute arrhythmia induced by inhaled chloroform – epinephrine rabbits model.Results The hypoxia tolerance in mice and myocardial hypoxia tolerance significantly improved by Zhurihen dripping pill, and prolonged the survival time in mice.The serum level of LDH, CK, SOD and MDA significantly improved, and protected the impaired myocardial cell in rats.The latent period of arrhythmia was significantly prolonged and duration of arrhythmia was shortened in rabbits.Conclusion Zhurihen dropping pills an improve myocardial oxygen deficiency, anti-arrhythmia effect and protect myocardial cells.
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This study was aimed to preliminary explore the anticancer activity, immunomodulatory effects and antimi-crobial activity of Calcitum ashing agent. Serum pharmacological method was used to prepare the serum containing Calcitum ashing agent. MTT method was used to observe the cell proliferation rate of S180 ascites tumor cells and human esophageal cancer cells cultured with the serum containing Calcitum ashing agent in vitro. In addition, this method was also used to observe the mouse spleen lymphocyte transformation rate. The Oxford Cup method was used to determine the antibacterial activity on Calcitum ashing agent in vitro. The results showed that serum containing Calcitum ashing agent inhibited the cell proliferation rate of S180 and esophageal cancer cells. Meanwhile, it had a synergistic effect with 5-fluorouracil (5-FU) on the inhibition of human esophageal cancer cells. The serum contain-ing Calcitum ashing agent induced the reduction in the mice spleen lymphocytes transformation rate. But it inhibited the reduction in the spleen lymphocytes transformation rate induced by 5-FU. The decoction with the concentration higher than 0.25 g·mL-1 had significantly inhibited the proliferation of bacteria. It was concluded that Calcitum ash-ing agent can inhibit the cell proliferation of S180 ascites tumor cells and human esophageal cancer cell in vitro. It can significantly reverse the reduction in the mice spleen lymphocytes transformation rate induced by immunode-pressant. However, the antibacterial activity still requires further study.
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AIM: To investigate the effect of different types of flavonoids, galangin (flavonol), hesperetin (flavonone) and hydroxysafflor yellow A (HYSA, chalcone), on cardiomyocytic injury induced by H2O2, and explore the possible signal pathways involved. METHODS: The cytotoxicity of different flavonoids was determined by MTT assay. Apoptosis rate was determined by flow cytometry (FCM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). Immunohistochemistry was used for detection of Bcl-2, Bax and Caspase-3 protein. RESULTS: It showed that each flavonoid did not have noticeable cytotoxicity at a concentration of 5 ?mol/L by MTT assay. Flavonoids at concentrations of 5, 15 and 30 ?mol/L significantly increased cell viability compared to model group induced by H2O2 (100 ?mol/L). Flavonoids also increased apoptosis rate and neorobiosis rate determined by FCM compared to model group. Galangin and hesperetin significantly decreased the apoptotic rate determined by TUNEL and the expression of Caspase-3 and increased the ratio of Bcl-2/Bax (P