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ObjectiveTo observe the effects of Sinisan on behaviors and NOD-like receptor protein 3 （NLRP3） inflammasomes of depressed rats induced by chronic unpredictable mild stress （CUMS） and further explore the anti-depressant mechanism of Sinisan. MethodFifty male rats were randomly divided into a normal group， a model group， an NLRP3 inhibitor （MCC950） group （10 mg·kg-1）， and low- （2.5 g·kg-1） and high-dose （5 g·kg-1） Sinisan groups， with 10 rats in each group. The depression model was induced by 42 d CUMS in rats except for those in the normal group. Drug intervention was performed on the 22nd day of modeling by gavage in the Sinisan groups and by intraperitoneal injection in the MCC950 group. Except for the MCC950 group， the remaining four groups received 10 mg·kg-1 physiological saline by intraperitoneal injection， while the rats in the model group， the normal group， and the MCC950 group were administered with 3 mL of physiological saline by gavage. Twenty-one days later， the sucrose preference test and open field test were performed. Western blot was used to detect the protein expression of NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， cysteinyl aspartate-specific protease-1 （Caspase-1）， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， ionized calcium-binding adapter molecule 1 （Iba1）， and CD68 in the hippocampus of rats in each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in the hippocampus of rats. Nissl staining and TUNEL were used to assess the pathological changes and apoptosis level in the hippocampal CA1 region of rats， respectively. ResultThe sucrose preference rate and consumption volume in the sucrose preference test， the total distance， the percentage of central movement distance， and the percentage of residence time in the open field test of rats in the model group were lower than those in the normal group （P<0.01）. Compared with the model group， the Sinisan groups and the MCC950 group showed improved depression-like behaviors， apoptosis level in the hippocampal CA1 region， and neuron loss to varying degrees. Sinisan could reduce the levels of IL-1β， IL-18， Bax， Iba1， and CD68 in the hippocampus （P<0.05， P<0.01）， increase the level of Bcl-2 （P<0.05， P<0.01）， and inhibit the protein expression of NLRP3， ASC， and Caspase-1 related to NLRP3 inflammasomes （P<0.05， P<0.01）. ConclusionSinisan can improve the depression-like behaviors and pathological damage of hippocampal neurons in CUMS-induced rats， and the mechanism may be related to the inflammatory response mediated by the NLRP3 inflammasome signaling pathway.
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Objective:To explore the mechanism of Bushen Huoxue prescription in regulating the related factors in phosphatidylinositol 3-kinase （PI3K）/ protein kinase B （Akt）/ mammalian target of rapamycin （mTOR） pathway and improving ovarian reserve function of rats with diminished ovarian reserve （DOR）. Method:Sixty DOR model rats were duplicated by Ataya method （intraperitoneal injection of cyclophosphamide） and then randomized into the model group， estradiol valerate （0.000 9 g·kg<sup>-1</sup>） group， and high- （33 g·kg<sup>-1</sup>）， middle- （16.5 g·kg<sup>-1</sup>） and low-dose （8.25 g·kg<sup>-1</sup>） Bushen Huoxue prescription groups， with 12 rats in each group. Another 12 healthy rats were classified into the blank control group. The rats in each group were given the corresponding drugs by gavage， while those in the blank control group and model group received the same volume of normal saline， once per day， for 14 successive days. After treatment， the ovarian tissue was stained with hematoxylin-eosin （HE） for observing the changes in quantities of primary follicles， mature follicles， and total follicles under a light microscope， followed by the detection of vascular endothelial growth factor （VEGF） expression in the ovarian tissue by immunohistochemistry. The protein expression levels of PI3K， Akt， mTOR， and cysteine-dependent aspartate-directed protease-3 （Caspase-3） in the ovarian tissue were assayed by Western blot， whereas the mRNA expression levels of PI3K， Akt， and mTOR were measured by Real-time polymerase chain reaction （Real-time PCR）. Result:As revealed by comparison with the blank control group， the quantities of mature follicles and total follicles in the ovarian tissue of model group were significantly reduced （<italic>P</italic><0.05， <italic>P</italic><0.01）. The protein expression levels of VEGF and Caspase-3 in the ovarian tissue were increased （<italic>P</italic><0.05）， while the protein and mRNA expression levels of PI3K， Akt， and mTOR were decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group， Bushen Huoxue prescription at the high and middle doses elevated the quantities of mature follicles and total follicles to varying degrees （<italic>P</italic><0.05， <italic>P</italic><0.01）， and VEGF increased most significantly in the middle-dose Bushen Huoxue prescription group （<italic>P</italic><0.05）. Caspase-3 in the low-， middle-， and high-dose Bushen Huoxue prescription groups and the western medicine group declined. The protein and mRNA expression levels of PI3K， Akt， and mTOR were up-regulated in the middle- and high-dose Bushen Huoxue prescription groups （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the levels in the middle-dose Bushen Huoxue prescription group were closer to those in the blank control group. Conclusion:Bushen Huoxue prescription effectively improves the ovarian reserve function of rats with DOR and increases the number of follicles possibly by up-regulating VEGF expression in the ovarian tissue， activating the PI3K/Akt/mTOR signaling pathway， and regulating the content of Caspase-3.
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Mast cells are widely distributed in various parts of the body, especially in the mucosal surface between the body and the external environment. Mast cell is one of the important immune cells and plays important roles in innate immunity, adaptive immunity and immune regulation. Previous researches have shown that excessive activation of mast cells is closely related to the development of allergic and inflammatory diseases such as asthma, allergic rhinitis, food allergies, acute and chronic itching. Mast cells infiltrate into the inflammation site and release various allergic mediators during the occurrence and development of these diseases. Therefore, termination of mast cell activation can be one of the effective methods for the treatment of allergic and inflammatory diseases, and receptors related to mast cell activation are potential targets for the development of anti-allergic drugs. There are many receptors related to mast cell activation, and the effects mediated by different receptors varied from each other. In the recent years, new mast cell receptors are being discovered, but there are not many literatures discussing the possible functions of these newly discovered receptors. This review aims to summarize the receptors involved in mast cell activation and classify related receptors according to their effects.
Subject(s)
Humans , Asthma , Allergy and Immunology , Hypersensitivity , Allergy and Immunology , Immunity, Innate , Inflammation , Allergy and Immunology , Mast Cells , Cell Biology , Allergy and ImmunologyABSTRACT
Objective To evaluate the clinical effectiveness of venipuncture using the fluorescent intravenous indwelling catheter in dark environment in dog experiments. Methods Six dogs were randomly divided into three groups of two dogs each, and each dog was subjected to 40 times of venipuncture performed on a foreleg with normal intravenous indwelling catheter and fluorescent intravenous indwelling catheter, respectively, under high(101 -105 lux), moderate (10.2-100 lux)or low(5 -10 lux)brightness conditions. The success rates and time consumption of those two procedures were analyzed and compared statistically. Results The success rate and time consumption for venipuncture using fluorescent and normal intravenous indwelling catheter under low brightness were 82.5% and(204.36 ± 13.13)s vs. 40.0% and(249.35 ± 17.98)s, those performed under moderate(simulating morning and dawn)brightness were 90.0% and(194.86 ± 8.60)s vs. 67.5% and(206.37 ± 9.70)s, all showed a significantly higher efficiency of the venipuncture using fluorescent intravenous indwelling catheter(P < 0.05). Conclusions The use of fluorescent intravenous indwelling catheter has the advantages of higher success rate and shorter time consumption for venipuncture under low and moderate brightness conditions.
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This paper was aimed to investigate the microRNA associated with multidrug resistance gene MDR1 of salvianolic acid A reversal in lung cance. Human lung cancer A549 cells were divided into normal control group and drug group, and the MDR1 expression levels were determined by real-time quantitative PCR. MicroRNA expression profiling of normal control group and drug group were detected by using the latest microRNA microarray. Quantitative RT-PCR was used to validate the differentially expressed miRNA. Forecast of miRNA associated with MDR1 multi-resistant genes of up-regulated miRNA. Experimental results showed that the dosage of MDR1 expression level significantly lowered compared with control group. The miRNA expression spectrum analyses of human lung cancer A549 cells to drug group and the control group were detected by microRNA microarray, 426 differentially expressed miRNA were screened out. Then target prediction were performed for difference up-expression of miRNA and found that there were four obvious increase of miRNA associated with MDR1 multi-resistant genes. Real-time quantitative PCR for 4 microRNA verification, the results were consistent with the chip. So the author considered that salvianolic acid A down lung cancer multidrug resistance gene MDR1 is likely to be affected by the miRNA expression and regulation of target genes, to further clarify the traditional Chinese medicine to reverse multi-drug resistant mechanism provides the experimental basis.
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Fifteen tissues of 4-year-old fruit repining stage Jilin ginseng were chosen as materials, six kinds of monomer saponins (ginsenosides Rg1, Re, Rb1, Rc, Rb2 and Rd) content in 15 tissues was measured by HPLC and vanillin-sulfuric acid method. The relative expression of FPS, SQS, SQE, OSC, β-AS and P450 genes in 15 tissues was analyzed by real-time PCR. The correlations between ginseng saponin content in 15 tissues of Jilin ginseng and biosynthetic pathway -related genes were obtained. The results showed that was a synergistic increase and decrease trend of positive linear correlation among six kinds of monomer saponin content, and there was a significantly (P < 0.01) positive correlation between monomer saponin content and total saponins content. Monomer saponin content and 6 kinds of enzyme gene correlation were different. Biosynthesis of ginseng total saponins and monomer saponin were regulated by six kinds of participation ginsenoside biosynthesis enzyme genes, the expression of these six kinds of genes in different tissues of ginseng showed collaborative increase and decrease trend, and regulated biosynthesis of ginseng ginsenoside by group coordinative manner.
Subject(s)
Drugs, Chinese Herbal , Gene Expression Profiling , Panax , Chemistry , Genetics , Metabolism , Plant Proteins , Genetics , Metabolism , Plant Structures , Chemistry , Genetics , Metabolism , Plants, Medicinal , Chemistry , Genetics , Metabolism , Saponins , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore ginseng fermentation process by Lactobacillus plantarum, and to make part of total saponins transformed into more reactive ginsenoside Rd.</p><p><b>METHOD</b>Microbial fermentation was carried out by still dark culture. Total saponins were extracted by Soxhlet extraction, and determined by UV visible spectrophotometry with colours reaction by vanillin-sulfuric acid. Ginsenoside Rd was determined by HPLC method.</p><p><b>RESULT</b>The fermentation process was: MRS medium, 35 degrees C, pH 5.0, cultured for 2 days. The content of total saponins was inhance 32%, and the content of ginsenoside Rd was increased 4.864 mg x g(-1).</p><p><b>CONCLUSION</b>The fermentation system's process was reasonable, and it's suitable for mass production, important significance for ginsenoside microbial transformation.</p>
Subject(s)
Biotransformation , Culture Media , Chemistry , Metabolism , Fermentation , Ginsenosides , Chemistry , Metabolism , Hydrogen-Ion Concentration , Lactobacillus plantarum , Chemistry , Metabolism , Molecular StructureABSTRACT
<p><b>OBJECTIVE</b>To investigate whether the neural representations underlying alternating two acupoint combinations (ACs) are the same or not.</p><p><b>METHODS</b>In this functional magnetic resonance imaging study, two sets of analgesia ACs were utilized, including Waiguan (TE5) and Qiuxu (GB40) for Group A, and Neiguan (PC6) and Taichong (LR3) for Group B, which are the most commonly adopted in clinical treatment. Each group had 20 healthy subjects. An experimental design was proposed, which consisted of a pre-needling resting phase, a needling phase and a post-needling resting phase. This paradigm optimally mimics the clinical protocol as well as focuses on both the stimulation and the resting periods. The results were subjected to general linear model analysis, conjunction analysis and the functional connectivity analysis.</p><p><b>RESULTS</b>The rostral anterior cingulated cortex was engaged in the initiative period of the acupuncture effect in both groups, and it was chosen as the seed region for the functional connectivity analysis for the following resting period. The results showed that several shared brain regions were involved in both groups, in particular the insula, amygdala and hypothalamus. Moreover, significant differences were located at the posterior cingulated cortex as revealed by a two sample -test (P<0.05, corrected). Other regions showed no significant differences. This finding was further supported by the spatial correlation analysis that the two groups were significantly correlated (r =0.51, P<0.01).</p><p><b>CONCLUSION</b>This preliminary research helps us understand the neurophysiological mechanisms of acupuncture when following clinical guidelines on ACs, as well as provides an important opportunity to develop better treatment strategies for reducing, or even preventing pain.</p>
Subject(s)
Female , Humans , Male , Young Adult , Acupuncture Points , Acupuncture Therapy , Gyrus Cinguli , Physiology , Magnetic Resonance Imaging , Nerve Net , Physiology , Nervous System Physiological Phenomena , Psychophysics , Rest , Signal Processing, Computer-Assisted , Statistics as TopicABSTRACT
Objective: To examine the biological accumulation of total ginsenosides and their monomers, and determine their relationships with the expression of squalene synthase (SQS) and squalene epoxidase (SQE) genes that are involved in the ginsenoside biosynthetic pathway in different organs of Panax quinquefolius. Methods: Fourteen organs of four year-old P. quinquefolius were used as materials. Total ginsenosides were extracted using the Soxhlet ginsenoside extraction method, and the contents of total ginsenosides and their monomers Rg1, Re, Rb1, Rc Rb2 and Rd in the organs were determined by the Vanillin-sulfuric Colorimetry and HLPC methods, respectively. The expressions of the SQS and SQE genes in the organs were profiled by real-time quantitative PCR. Results: The biological accumulation of total ginsenosides and each of their monomers varied significantly (P<0.01) in different parts of P. quinquefolius.Except for ginsenoside monomer Rb 2, there were significantly positive correlations between total ginsenoside and monomers Re, Rg1, Rb1 and Rd (P<0.01). The expressions of both SQS and SQE genes were extremely significantly different among the 14 plant parts (P<0.01) and significantly positively correlated with the biological accumulation of total ginsenoside and monomers, Re, Rg 1, Rb1 and Rd (P<0.05). Conclusion: The results indicate that the SQS and SQE genes play the important roles in the biosynthesis of total gingenosides and their monomers.