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Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.
ABSTRACT
Unlike adult mammalian heart, zebrafish heart has a remarkable capacity to regenerate after injury. Previous study has shown Notch signaling activation in the endocardium is essential for regeneration of the myocardium and this activation is mediated by hemodynamic alteration after injury, however, the molecular mechanism has not been fully explored. In this study we demonstrated that blood flow change could be perceived and transmitted in a primary cilia dependent manner to control the hemodynamic responsive klf2 gene expression and subsequent activation of Notch signaling in the endocardium. First we showed that both homologues of human gene KLF2 in zebrafish, klf2a and klf2b, could respond to hemodynamic alteration and both were required for Notch signaling activation and heart regeneration. Further experiments indicated that the upregulation of klf2 gene expression was mediated by endocardial primary cilia. Overall, our findings reveal a novel aspect of mechanical shear stress signal in activating Notch pathway and regulating cardiac regeneration.
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Objective To study the effect of Angelicin on proliferation activity and anti-aging related protein expression of human HDF cells and its mechanism. Methods According to the random number table method, the cells were divided into blank group, model group, estradiol group, Angelicin group, estrogen receptor antagonist+estradiol group, estrogen receptor antagonist+Angelicin group, and P38 pathway blocker group. Different groups were given the according drugs respectively for 24 h. Except the blank group, all the groups of cells were given UVB irradiation with a dose of 150 mJ/cm2. The MTT assay was used to detect cell proliferation rate. The Western blot was used to detect the expression levels of COLⅠ, MMP-1, ERβ, P38 and p-P38 in cells, and the MMP-1 mRNA expression was detected by real-time fluorescent quantitative PCR.Results Compared with the model group, the proliferation rate of HDF cells significantly increased in Angelicin(10,1,0.1 and 0.01 μmol/L groups)(P<0.01);The protein expression of COLⅠ (0.326 ± 0.006 vs. 0.176 ± 0.007),ERβ(0.281 ± 0.011 vs.0.143 ± 0.006)significantly increased(P<0.01),and the expression of MMP-1(0.256 ± 0.006 vs.0.395 ± 0.006)and p-P38(0.224 ± 0.003 vs.0.318 ± 0.005)significantly decreased (P<0.01) in Angelicin 10 μmol/L group. Compared with 10 μmol/L Angelicin group, the protein expression of estrogen receptor antagonist+Angelicin group ERβ(0.120 ± 0.007 vs.0.281 ± 0.011)significantly decreased and MMP-1mRNA(1.377 ± 0.012 vs.1.024 ± 0.010)significantly increased(P<0.01).Conclusions The Angelicin may degrade MMP-1 through the ER-P38 MAPK signaling pathway,and then promote collagen synthesis, to achieve the purpose of prevention and treatment of photoaging.
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Objective To study the effect of psoralen isoflavone on the treatment of PC12 cells injured by Aβ and the mechanism on the effect of the psoralen isoflavones on the expression of related proteins. Methods The PC12 cells were divided into blank group, model group, E2 group, and psoralen isoflavones group by random number table method, In addition to the blank group the rest of each group culture medium were added 20μmol/L of Aβ25-35 modeling, The E2 group was added to the 10-3μmol/L oestrogen and psoralen isoflavones group for the intervention of 102-10-6 μmol/L.The proliferation rate of PC12 cells was detected by MTT assay, and the expression of APP, BACE1, ERβ, p-ERK and Aβ protein was detected by Western Blot. Results Compared with the model group, the proliferation of PC12 cells induced by 10-1μmol/L of psoralen isoflavone increased (101％ vs. 52％, P<0.01); The expression of p-ERK (0.751± 0.066 vs. 0.364 ± 0.015), ERβ(0.756 ± 0.105 vs. 0.337 ± 0.045) increased significantly (P<0.01); APP (0.382 ± 0.039 vs. 0.479 ± 0.015), BACE1 (0.517 ± 0.024 vs. 0.622 ± 0.029), Aβ (0.430 ± 0.032 vs. 0.581 ± 0.030) expression amount were significantly lower (P<0.05). Conclusions Psoralen isoflavones have a certain therapeutic effect on PC12 cells injured by Aβ.
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BACKGROUND: Open facture is easy to induce infection, which is an urgent problem in clinic. Establishing a reliable animal model of open fracture with infection is of great significance for drug and instrument development and application. OBEJCTIVE: To develop an open fracture with infection model in New Zealand white rabbits, and to identify the available number of bacteria that can cause infection. METHODS: The amount of bacteria was determined by establishing open fracture structure and verifying the concentration of bacterial colonies. Thirty New Zealand white rabbits were randomly divided into control group and four experimental groups, and a transverse fracture at the middle part of tibia was established in all rabbits,followed by the injection of 1 mL of normal saline or 1mL of Staphylococcus aureus suspension at the concentrations of 1×105, 1×106, 5×106, and 1×107CFU/mL. Afterwards, the optimal concentration of 1 mL of bacteria liquid causing infection was determined by gross observation, body temperature analysis and body mass measurement, white blood cell and C-reactive protein detection, bacterial culture and pathological observation. RESULTS AND CONCLUSION: Rabbits in the 5×106CFU/mL group were all infected and had higher survival rate. In the 1×105and 1×106CFU/mL groups, some rabbits showed no infection. One rabbit died due to infection in the 1×107CFU/mL group. In summary, the reliable infection model of open fracture can be induced by injected with 1 mL of Staphylococcus aureus at the concentration of 5×106CFU/mL in New Zealand white rabbits, which can be used as an effective model to guide drugs and instruments related anti-infective research.
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Objective To study the rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB. Methods The potential of cell proliferation of the different concentrations of rhubarb phenol and P38 inhibitor (SB203580) on human skin HaCaT was detected by MTT method. The cells was divided into the control group, the model group, the rhubarb phenol group, the SB203580 group by random number table method after the 24 h incubation. The 10-6 mol/L rhubarb phenol and 10-7 mol/L SB203580 were added to the rhubarb phenol group and SB203580 group for 24h, The competence of cultured cell proliferation which was irradiated with UVB of intensity of 0.61mW/cm2, mutiply time of 7 min and distance of 10 cm for 24 h except the control group; Western Blot method detected rhubarb phenol and P38 inhibitor of the influence of P38, P-P38, TNF-α, IL-6 protein. Results Compared with control group, the cell proliferation in UVB group significantly reduced (P<0.01); Compared with UVB group, the expression of the P-P38 (0.419 ± 0.029, 0.398 ± 0.015 vs. 0.497 ± 0.051), TNF-α (0.435 ± 0.025, 0.411 ± 0.021 vs. 0.509 ± 0.040) and IL-6 (0.457 ± 0.027, 0.432 ± 0.018 vs. 0.478 ± 0.036) in rhubarb phenol and P38 inhibitor group significantly reduced (P<0.01). Conclusions The rhubarb phenol and P38 inhibitor could significantly suppress HaCaT cells light aging, and its mechanism may be related with inhibiting P38 signaling pathways, and inhibiting the secretion of inflammatory cytokines.
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Objective To study the pinoresinol diglucoside (PDG) on gene regulation role of ESF-1 cells in collagen secretion, to reveal PDG repair mechanisms on scalded skin.Methods The cells cultured in vitro were divided into the control group, the estradiol group and the three different PDG doses groups. The concentration of the high, medium and low dose groups were 100, 10, 1μmol/L, and that of estradiol group were 10-3μmol/L. The activity of proliferation was detected by MTT. Then collagen type I (Col I), collagen typeⅢ (ColⅢ), tissue inhibitors of metalloproteinase 1 (TIMP-1), tissue inhibitors of metalloproteinase 2 (TIMP-2) and matrix metalloproteinase 1 (MMP-1) expression levels of mRNA after administration of cells were detected by RT-PCR.Results Compared with the control group, the proliferation of ESF-1 cells (0.559 ± 0.027, 0.552 ± 0.034vs. 0.489 ± 0.027,P<0.05) in the estradiol and medium-dose PDG was significantly higher. The expression level of mRNA of ColⅠ(0.958 ± 0.021, 0.929 ± 0.031, 0.916 ± 0.015vs. 0.844 ± 0.022), ColⅢ (0.783 ± 0.038, 0.918 ± 0.021, 0.855 ± 0.017vs. 0.678 ± 0.024), TIMP-1 (0.939 ± 0.025, 0.889 ± 0.036, 0.853 ± 0.015 vs. 0.780 ± 0.023), TIMP-2 (0.507 ± 0.024, 0.655 ± 0.037, 0.572 ± 0.025vs. 0.405 ± 0.062) in the estradiol, low-, medium-dose PDG groups were significantly higher than those in the control group (P<0.05 or P<0.01). Besides, the MMP-1 (0.343 ± 0.038, 0.407 ± 0.046, 0.435 ± 0.037vs.0.519 ± 0.041) mRNA expression level in the middle and low dose PDG groups significantly decrease (P<0.05 orP<0.01). Conclusions The PDG could enhance the activity of ESF-1 cell proliferation, increase the expression of related collagen and tissue inhibitor of metalloproteinases and inhibit that of matrix metalloproteinases to repair scalded skin.
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This study was aimed to investigate the effects of Bu-Yang Huan-Wu (BYHW) decoction to the learning and memory ability of APP/PS1 double transgenic mice through two behavior tests,in order to observe the Alzheimer's disease (AD) treatment effect of BYHW decoction.A total of 60 APP/PS1 double transgenic mice were randomly divided into five groups,which were the model group,the hydrochloride group,the high-,middle-,and low-dose BYHW decoction group,with ten rats in each group.The C57BL/6J mice were used as the control group.After 30 days of drug administration,all mice were subjected to behavior tests,including new object recognition experiment and step-through task.Western blot was used to detect changes of inflammatory factors,including IL-6 and TNF-α,in hippocampus of mice.The results showed that compared with the model group,the high-and middle-dose BYHW decoction can significantly decrease inflammatory factors of IL-6 and TNF-α expression in hippocampus of APP/PS1 mice.It can also obviously improve the learning and memory ability of APP/PS1 mice.It was concluded that BYHW decoction can significantly decrease the inflammatory factor expression in APP/PS 1 double transgenic AD model mice.It can obviously improve its learning and memory ability,which can be used in the treatment of AD.
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Objective To study the effect of Schisandra chinensis on the treatment of glomerular nephritis in rats by gas chromatography mass spectrometry (GC-MS).Methods The rats were randomly divided into normal group, model group, and the treatment group, 10 rats in each group. We employed the C-BSA injuction to induce situ immune complexes nephritis rate model. Then 24 hour urinary protein was detected after 12 days and a urine metabonomic technology combined with pattern recognition method was used to identify the network analysis, which focuses on the key metablites and enzmys with glomerulonephritis disease.Results After 12 days, compared with the control group, the 24 h urinary protein content (9.24 ± 2.69 mg vs. 76.04 ± 18.88 mg) significantly decreased (P<0.01). Urine metabonomicsi dentified 15 endogenous biomarkers, which included unsaturated fatty acid biosynthesis, pyruvate metabolism and glycolysis metabolism. These metabolic pathways are associated with kidney diseases.Conclusion The effect of Schisandra chinensis on glomerular nephritis may treated by regulating the metabolism of arachidonic acid and fatty acid in four.
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Objective To investigate the effects of genistein on the mRNA expressions of collagen (Col), matrix metalloproteinase (MMP ) and tissue inhibitor of matrix metalloproteinase (TIMP) in human embryonic skin fibroblasts (CCC-ESF-1).MethodsThe cultured CCC-ESF-1cells were divided into a black control group, an estradiol group and genistein groups of different doses. The mRNA expressions of ColⅠ, ColⅢ , MMP-1, TIMP-1 and TIMP-2 were detected by RT-PCR.Results Compared with the black group, estradiol and medium dose of genistein (0.451 ± 0.037, 0.446 ± 0.047vs.0.385 ± 0.061, allP<0.05) could promote the proliferation of the CCC-ESF-1 cells, estradiol and medium dose of genistein could up-regulate the mRNA expressions of ColⅠ (0.960 ± 0.012, 0.929 ± 0.015vs.0.812 ± 0.014, allP<0.01), ColⅢ (0.892 ± 0.009, 0.824 ± 0.022vs.0.768 ± 0.025, allP<0.01), TIMP-1 (0.841 ± 0.023, 0.838 ± 0.053vs.0.751 ± 0.027, allP<0.01) and TIMP-2 (0.456 ± 0.017, 0.448 ± 0.036vs.0.381 ± 0.029, allP<0.01), and down-regulate MMP-1 mRNAexpression (0.398 ± 0.043, 0.402 ± 0.044vs.0.525 ± 0.006, allP<0.01).Conclusions Genistein could promote the proliferation of the CCC-ESF-1 cells, and that may be related with up-regulating the mRNA expressions of ColⅠ, ColⅢ , MMP-1, TIMP-1and down-regulating MMP-1 mRNA expression.
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<p><b>OBJECTIVE</b>To explore the relationship between aging and erectile function changes in rats in order to establish a rat model of aging-related erectile dysfunction (ED).</p><p><b>METHODS</b>Eighty male Wistar rats were equally divided into four age groups (3-, 6-, 12- and 18-month) and treated with intragastric administration of sildenafil citrate (Sn) for penile erection tests. Twenty 3-month-old female Wistar rats were randomized to four groups as oestrous rat models. We recorded the rate and frequency of penile erections of the male rats in different age groups.</p><p><b>RESULTS</b>The rates of penile erection were 85%, 75%, 40% and 30% and erectile frequencies were 2.27 +/- 0.80, 2.00 +/- 0.61, 1.40 +/- 0.51 and 1.29 +/- 0.49 in the 3-, 6-, 12- and 18-month rats, respectively, with statistically significant differences among different age groups (P < 0.01). And their erectile function exhibited a tendency to decrease with the increase of age. Besides, comparison of the 3-month with the 6-, 12- and 18-month groups showed significantly reduced erectile function in the 18-month rats (P < 0.05) but no remarkable difference between the 3-month and the 6- and 12-month groups (P > 0.05).</p><p><b>CONCLUSION</b>Aging is one of the main risk factors of rat erectile dysfunction, and 18-month-old male rats are qualified for the establishment of the rat model of aging-related erectile dysfunction.</p>
Subject(s)
Animals , Male , Rats , Aging , Physiology , Erectile Dysfunction , Models, Animal , Penile Erection , Physiology , Rats, WistarABSTRACT
Plantamajoside is one of the main bioactive compounds in Plantaginis Herba A TLC method was developed identification of plantamajoside in 11 Plantaginis Herba samples using silica gel G as coating substance and a mixture of ethyl acetiate methanol-formic acid-water (18: 3 : 1.5 : 1) as a developing solvent, the established TLC condition displayed a very well separation on the chromatogram of tested Plantaginis Herba samples and the marker compound plantamajoside showed as a distinct light-blue fluorescence spot observed under UV 365 nm. Using the HPLC method, plantamajoside was separated at 30 degrees C on a Promocil C18, (4.6 mm x 250 mm, 5 microm) column with acetonitrile-0.1% formic acid (17:83) as the mobile phase. The detection wavelength was set at 330 nm and the flow rate was 1 mL x min(-1). The calibration curve of plantamajoside displayed ideal linearity over the range of 0.0499-11.9664 microg (r = 0.9999), and the average recovery of plantamajoside was 100.6% with a RSD of 2.7%. The contents of plantamajoside were in the range of 0.067%-1.80% in Plantaginis Herba The established TLC identification and HPLC were sensitive, reliable and repeatable, which can be applied for the quality evaluation and standard criteria of Plantaginis Herba.
Subject(s)
Asteraceae , Chemistry , Catechols , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Glucosides , Reproducibility of ResultsABSTRACT
<p><b>OBJECTIVE</b>The research on flavonoid in the recent years is extensive, such as Soybeans flavone, Baicalensis flavone, Epimedium flavone. Experiments show the effects of flavonoid is closely related to human health. There are a lot of reports about Ginkcobiobal flavone. In order to make further progress research on ginlcgobiobal flavone, we sum up the articles and reports on ginlcgobiobal flavone in the recent years.</p><p><b>METHOD</b>To searche the articles about ginlcgobiobal flavone studies in the past five years.</p><p><b>RESULT</b>Ginlcgobiobal flavone is not only a vasodilator, but also has the effects of anti-inflammation, analgesia, lowering blood lipids, preventing senile and inhibiting tumor, treating leukaemia, regulating gene and biotransformation.</p><p><b>CONCLUSION</b>Ginlcgobiobal flavone has the potential value for drug research and development.</p>