Objective To explore the potential effects of endothelial progenitor cells (EPCs)-angiogenesis on mechanism of alleviating cognitive dysfunction in rats subjected to cerebral ischemia-reperfusion (I/R) injury. Methods A total of 121 male Sprague–Dawley (SD) rats were randomly divided into four groups:Sham group (n=31), focal I/R(MCAO, 0.9%saline 10μL, n=30) group, MCAO+Vehicle (sodium azide, 0.1%Vehicle 10μL, n=30) group and MCAO+HPX (1.86 g/L HPX 10μL, n=30) group. The modified neurological severity scores (mNSS) was carried out to determine neurological function deficit after I/R. Morris water maze (MWM) was carried out to assess learning and memory abilities after I/R. The circulating EPCs after I/R were counted by flowcytometry (FCM) combined with double-immunofluorescence staining of CD34 and CD133. Angiogenesis in rat penumbra cortex after I/R was assessed by immunohistochemical technique combined with immunofluorescent chromogenic detection of CD31 and vWF. Results Compared with sham group, the mNSS scores, the escape latency and the circulating EPCs count were increased after I/R, the time percentage spent in target quadrant was reduced, and the new vessel density in penumbra cortex was increased after I/R in MCAO group (P < 0.05 respectively). There were no significant differences in mNSS score, the escape latency, the time percentage spent in target quadrant, the circulating EPCs count and the new vessel density in penumbra cortex between MCAO group and MCAO+Vehicle group ( P>0.05). The mNSS score and the escape latency were significantly decreased, the circulating EPCs count and new vessel density in penumbra cortex were significantly increased after I/R in MCAO+HPX group compared with those of MCAO+Vehicle and MCAO group (P<0.05). Conclusion EPCs-angiogenesis signaling plays positive effects on HPX alleviating cognitive dysfunction in rats subjected to focal cerebral ischemia reperfusion injury.

This study was to investigate the protective effects of Jinqi Jiangtang tablets on diabetic complications of cardiovascular disease in Drosophila. The effects of Jinqi Jiangtang tablets on hypolipidemia and hypoglycemia by high fat diet(HFD)induced model organism Drosophila with the content of triglyceride and trehalose in Drosophila as indexes, were investigated. And the Drosophila heart function was detected by high speed EM-CCD; cell signaling pathways were detected by Western blot and RT-PCR. The results showed that high fat diet could induce the appearance of hyperglycemia and hyperlipidemia on Drosophila model. Jinqi Jiangtang tablets could significantly reduce triglyceride and trehalose and protect heart function of Drosophila induced by high fat diet. Jinqi Jiangtang tablets could also increase the expression of 4ebp mRNA, while decreasing p-4EBP protein and pepck mRNA expression. This study demonstrated that Jinqi Jiangtang tablets have a protective effect on HFD-induced dyslipidemic-diabetic and cardiac dysfunction, which may be related to mTOR/4EBP pathway.

Objective:To evaluate the role of heme oxygenase-1 (HO-1) in hemopexin (HPX)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in rats.Methods:One hundred and eighty male Sprague-Dawley rats, aged 7-8 weeks, weighing 250-280 g, were divided into 5 groups ( n=36 each) using a random number table method: sham operation group (group SH), I/R group, vehicle group (group V), HPX group, and HPX plus HO-1 specific inhibitor ZnPPIX(HZ group). Local cerebral I/R was produced by middle cerebral artery occlusion for 120 min followed by reperfusion in anesthetized rats.In I/R, V, HPX and HZ groups, 0.9% normal saline 10 μl, 0.1% NaN 3 10 μl, 1.86 g/L HPX (diluted to 10 μl in 0.1% NaN 3 solution), and 1.86 g/L+ ZnPPIX 20 μmol/L (diluted to 10 μl in 0.1% NaN 3 solution) were injected through the lateral ventricle, respectively, immediately after onset of reperfusion.Morris water maze test was used to detect the cognitive function on day 1 before ischemia and day 2 of reperfusion.Rats were sacrificed, brains were removed and brain tissues were obtained for determination of the permeability ratio of Evans blue (EB), brain water content, expression of VE-cadherin in ischemic penumbra (by Western blot), and expression of angiopoietin-1 (Ang1) mRNA and Ang2 mRNA (by real-time polymerase chain reaction). Ang1 mRNA/Ang2 mRNA ratio was calculated.CD31/vWF double labeling immunofluorescence was used to detect the density of neovascularization in hippocampal tissues in the ischemic penumbra. Results:Compared with SH group, the escape latency was significantly prolonged at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was decreased, the frequency of crossing the platform was decreased, the permeability ratio of EB in brain tissues was increased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were decreased in I/R, V, HPX and HZ groups ( P<0.05). Compared with I/R group, the escape latency was significantly shortened at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was increased, the frequency of crossing the platform was increased, the permeability ratio of EB in brain tissues was decreased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were increased in HPX group ( P<0.05), and no significant change was found in the parameters mentioned above in V and HZ groups ( P>0.05). Compared with HPX group, the escape latency was significantly prolonged at 2-7 days of reperfusion, the percentage of time of staying at the target quadrant was decreased, the frequency of crossing the platform was decreased, the permeability ratio of EB in brain tissues was increased at day 7 of reperfusion, and the brain water content, Ang1 mRNA/Ang2 mRNA ratio, expression of VE-cadherin and density of neovascularization were decreased in HZ group ( P<0.05). Conclusion:HO-1 is involved in HPX-induced reduction of cerebral I/R injury in rats.

Fibrosis refers to the final outcome of damage in multiple-type tissue and the imbalance of tissue repair especially in the process of chronic inflammatory response diseases.Fibrosis can occur in various organ tissues.Its continuous progression may lead to organ dysfunction and failure,which is a huge threat to human health.Traditional Chinese medicine has significant therapeutic effects in preventing and treating fibrosis.Due to its characteristics of multiple components,pathways,and targets,it has become a hot research topic in the field of fibrosis.Astragali Radix,a Chinese medicinal for supplementing qi,is the root of Astragalus membranaceus(Fisch.)Bge.var.mongholicus Hisao or Astragalus membranaceus(Fisch.)Bge.It has the effects of replenishing qi and elevating yang,generating fluid and nourishing blood,expelling toxin and draining pus,astringing sore and promoting granulation.It has found that Astragali Radix contains many chemical components such as polysaccharides,saponins,and flavonoids,which have good anti-inflammatory and antioxidant effects.Astragali Radix can effectively intervene in the fibrosis process of multiple organ tissues such as the heart,kidney,liver,and lung.Therefore,this article reviews the anti-fibrotic effects and mechanisms of Astragali Radix and its chemical components,hoping to provide ideas and references for the development and utilization of Astragali Radix.

OBJECTIVE To qualitatively analyze the chemical constituents from Tianzhi granules and their constituents absorbed into blood， and to provide reference for elucidating pharmacodynamic material basis of Tianzhi granules. METHODS UPLC-ESI-Q-TOF-MS/MS was adopted. The analysis was performed on an Agilent Eclipse Plus C18 column with mobile phase consisted of 0.5% formic acid solution-acetonitrile （gradient elution） at the flow rate of 0.3 mL/min； the column temperature was 40 ℃ ；the injection volume was 10 μL. Mass spectrometry was applied for the qualitative analysis under positive ionization mode and ESI ion source. Data were collected with MS-DIAL4.60， and then chemical constituents of the extract from Tianzhi granule （by 0.5% methanol） were analyzed by comparing with relevant literature， SciFinder， PubChem， MassBank， TCMSP， TCM-ID and other databases. The blank serum， administered serum and Tianzhi granule extract were compared to analyze the constituents absorbed into the blood. RESULTS One hundred compounds were preliminarily identified from Tianzhi granules， including 46 flavonoids， 8 organic acids， 8 alkaloids， 6 terpenes， 6 coumarins， 2 quinones， 1 steroids， 7 glycosides and 16 others. Based on it， 10 prototype constituents absorbed into blood were identified preliminarily， including genistein， melatonin A， chrysin-7-O- β-glucuronide， apigenin-7-O-glucuronide， wogonin， 6-O-methylbaicalin are flavonoids， 2-hydroxyquinoline and isonacolline are alkaloids， 7-hydroxycoumarin is coumarins，1-indanol is others. CONCLUSIONS 2-hydroxyquinoline， 7-hydroxycoumarin， genistein， melatonin A， isonocolline， chrysin-7-O-β-glucuronide， apigenin-7-O-glucuronide， wogonin and 6-O-methylbaicalin may be the pharmacodynamic material basis of Tianzhi granules.

OBJECTIVE To explore the effect mechanism of ethanol extract from Atractylodes macrocephala （EEAM） on microglial phagocytosis and degradation of amyloid β （Aβ） based on peroxisome proliferator-activated receptor γ （PPAR- γ） signaling pathway. METHODS Taking neuromicroglial cell BV2 as subjects， confocal microscopy was used to observe the effects of EEAM （0.3， 0.4， 0.5 mg/mL， similarly hereinafter） on phagocytosis and degradation of Aβ in microglia. Human embryonic kidney cell HEK293 was used to investigate the effects of EEAM on luciferase transcriptional activity of PPAR-γ. The effect of EEAM on nuclear translocation of PPAR-γ was investigated by immunofluorescence. Alzheimer’s disease BV2 cell model was induced by Aβ1-42， and quantitative polymerase chain reaction was used to investigate the effects of EEAM on mRNA expressions of PPAR-γ downstream target genes （Lxra， Lxrb， Abca1， Abcg1， Cd36， Sra and Apoe）. RESULTS The results of Aβ uptake experiment showed that after the intervention of medium and high doses of EEAM， fluorescence intensity of Aβ in BV2 cells increased significantly （P＜0.05）. The degradation experiment of Aβ showed that after the intervention of medium and high doses of EEAM， fluorescence intensity of Aβ in BV2 cells decreased significantly （P＜0.05）. After the intervention of different doses of EEAM， luciferase transcriptional activity of PPAR-γ in HEK293 cells increased significantly （P＜0.05）； fluorescence intensity of PPAR-γ in BV2 cells and nuclei （except for low-dose group） increased significantly （P＜0.05）. mRNA expressions of Lxra， Lxrb， Abca1， Abcg1， Cd36， Sra and Apoe in BV2 cells were increased significantly （P＜0.05）. CONCLUSIONS EEAM can promote the uptake and degradation of Aβ in microglia by activating PPAR-γ signaling pathway， thus improving Alzheimer’s disease.

ObjectiveTo investigate the effect of Yishen Tongluo prescription （YSTLP） on apoptosis of renal tubular epithelial cells and explore the mechanism based on endoplasmic reticulum stress pathway of protein kinase R-like endoplasmic reticulum kinase （PERK）/activating transcription factor 4 （ATF4）/transcription factor C/EBP homologous protein （CHOP）. MethodThe db/db mice were randomly divided into model group， valsartan group （10 mg·kg^-1）， and low， middle， high-dose YSTLP groups （1， 2.5， 5 g·kg^-1）. Samples were collected after eight weeks of drug intervention. In addition， db/m mice in the same litter served as the control group. Human renal tubular epithelial cells （HK-2） were cultured in vitro and divided into the control group， advanced glycated end-product （AGE） group， and AGE + low， middle， and high-dose YSTLP groups （100， 200， 400 mg·L^-1）. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect the apoptosis rate of HK-2 cells. Methylthiazolyldiphenyl-tetrazolium bromide （MTT） assay was conducted to detect the viability of HK-2 cells. Calcium fluorescence probe staining and luciferase reporter gene method were adopted to detect the luciferase activity of folded protein response element （UPRE） and endoplasmic reticulum stress. Immunohistochemical （IHC） analysis was carried out to measure the protein expressions of phosphorylated PKR （p-PERK）， CHOP， and ATF4. Real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression levels of CHOP and X-box binding protein 1 （XBP1） in mouse kidney and HK-2 cells. Western blot was used to detect the protein expression level of p-PERK， PERK， CHOP， ATF4， B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， and cleaved Caspase-3 in mouse kidney and HK-2 cells. ResultIn the cellular assay， HK-2 cell viability was significantly reduced， and the apoptosis rate was elevated in the AGE group compared with the control group （P<0.01）. The mRNA and protein expression levels of apoptosis-related factor Bcl-2 were significantly reduced （P<0.01）， and those of Bax were significantly increased （P<0.01）. The protein expression level of cleaved Caspase-3 was significantly increased （P<0.01）. Compared with the AGE group， YSTLP administration treatment resulted in elevated cell viability and reduced apoptosis rate （P<0.01）. The mRNA and protein expression levels of Bcl-2 were significantly elevated in a time- and dose-dependent manner （P<0.01）， and those of Bax were significantly reduced in a time- and dose-dependent manner. The protein expression level of cleaved Caspase-3 was significantly reduced in a time- and dose-dependent manner （P<0.01）. The intracellular Ca²⁺ imbalance and UPRE luciferase fluorescence intensity were increased in the AGE group compared with the control group （P<0.01）. The mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 were significantly increased （P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the AGE group， YSTLP effectively improved intracellular Ca²⁺ imbalance in HK-2 cells and decreased UPRE luciferase fluorescence intensity in a dose-dependent manner （P<0.01）. It reduced the mRNA levels of endoplasmic reticulum stress-related factors CHOP and XBP1 （P<0.01） and the protein expression levels of intracellular p-PERK， CHOP， and ATF4 in a dose- and time-dependent manner （P<0.01）. In animal experiments， the protein expression level of Bcl-2 was significantly reduced（P<0.01）， and that of cleaved Caspase-3 and Bax was significantly increased in the model group compared with the control group （P<0.05）. The protein expression level of Bcl-2 was dose-dependently elevated， and that of cleaved Caspase-3 and Bax was dose-dependently decreased in the YSTLP groups compared with the model group （P<0.01）. Compared with the control group， the mRNA expression levels of CHOP and XBP1 were significantly elevated in the model group （P<0.05， P<0.01）， and the protein expression levels of p-PERK， CHOP， and ATF4 were significantly increased （P<0.05）. Compared with the model group， YSTLP significantly decreased the mRNA expression levels of CHOP and XBP1 （P<0.01） and the protein expression levels of p-PERK， CHOP， and ATF4 （P<0.01）. ConclusionYSTLP can effectively inhibit endoplasmic reticulum stress and improve apoptosis of renal tubular epithelial cells， and its mechanism may be related to the regulation of the PERK/AFT4/CHOP pathway.

OBJECTIVE： To study the effects of Atractylodes macrocephala ethanol extract （AM） on life span of Caenorhabditis elegans（called N 2 nematode for short ），and to investigate its mechanism based on transcription factor SKN- 1/ nuclear factor E 2 related factor 2（Nrf2）. METHODS ：N2 nematode were divided into blank control group ，positive control group （100 μ mol/L curcumin，similarly hereinafter ），AM low-dose ，medium-dose and high-dose groups （100，200，300 μ g/mL， similarly hereinafter ）. The effects of AM on the life span （by average survival time ）of N 2 nematode under normal condition and oxidant stress condition （40 mmol/L H 2O2）as well as its effects on reproductive capability （by the number of filial generation ）of N2 nematode under normal condition were investigated . 700 μmol/L H2O2 was used to establish neuroblastoma cells N 2a oxidant stress model. Effects of positive control ，low-dose，medium-dose and high-dose of AM on the survival rate of model cells were detected by MTT method. After human embryonic renalepithelial cells 293T were transfected with Nrf 2-ARE plasmid ， the effects of positive control and AM on luciferase activity of Nrf2-ARE were detected by luciferase reporter gene method at low，medium and high dose for 24 h and at medium dose for 12，18 and 24 h. RT-PCR was used to detect the effects ofpositive control ，low-dose，medium-dose and high-dose of AM on the mRNA expression of downstream genes NQO- 1 and HO- 1 of Nrf 2 in N 2a cells as well as mRN A expression of en@hactcm.edu.cn downstream genes GCS- 1，GST-7，GST-10，HSP-60，HSP- 16.2 and SOD- 3 of SKN- 1 in N 2 nematode. RESULTS ：Compared with blank control group ，average survival time of N 2 nematode under normal and oxidant stress condition was significantly prolonged in positive control group and AM groups ；the number of filial generation on the first day （except for AM high-dose group ），the number of filial generation on the second day （except for AM low-dose group ） and the total number of filial generation （except for AM low-dose group ） were increased significantly（P＜0.05 or P＜0.01）. The survival rate of N 2a cells in positive control group ，AM medium-dose and high-dose groups were significantly higher than that of model group （P＜0.05 or P＜0.01）. Compared with blank control group ，Nrf2-ARE luciferase relative activity of 293T cells in positive control group and AM groups as well as Nrf 2-ARE luciferase relative activity of 293T cells in AM medium-dose group after different time of treatment were increased significantly （P＜0.01），in dose-dependent and time-dependent trend. Compared with blank control group ，mRNA relative expression of HO- 1 and NQO- 1（except for positive control group ），GCS-1（except for AM low-dose group ），GST-7（except for positive control group and AM low-dose group ）， GST-10 and HSP- 60（except for AM low-dose group ），HSP-16.2（except for positive control group and AM low-dose group ）and SOD-3 （except for positive control group and AM low-dose group ） were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：AM can prolong the life span of N 2 nematode under normal and oxidant stress condition and improve the its reproductive capacity ，the mechanism of which may be associated with the activation of SKN- 1/Nrf2 signaling pathway.

ObjectiveTo investigate the mechanism of salvianolic acid F （Sal F） in repairing the high glucose-induced injury in human kidney-2 （HK-2） cells via the B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）/cysteinyl aspartate-specific proteinase 3 （Caspase-3）/gasdermin-E （GSDME） pathway. MethodThe cell counting kit-8 （CCK-8） was used to measure the relative viability of HK-2 cells exposed to high glucose and different concentrations （2.5， 5， 10， 20 μmol·L^-1） of Sal F and the relative viability of HK-2 cells treated with Sal F for different time periods. The levels of lactate dehydrogenase （LDH） and interleukin-1β （IL-1β） in the supernatant of the cell culture were measured by the LDH assay kit and enzyme-linked immunosorbent assay （ELISA） kit， respectively. Flow cytometry combined with Annexin V-FITC/propidium iodide （PI） and Hoechst 33342/PI staining was employed to reveal the proportion of PI-positive HK-2 cells exposed to high glucose. Western blotting was employed to determine the protein levels of Bax， Bcl-2， cytochrome C， cysteinyl aspartate-specific proteinase （Caspase）-9， Caspase-3， and GSDME in the HK-2 cells exposed to high glucose and treated with Sal F. The 2，7-dichlorodihydrofluorescein diacetate fluorescence probe （DCFH-DA） and mitochondrial membrane potential assay kit （JC-1） were used to determine the production of reactive oxygen species （ROS） and the mitochondrial membrane potential in the HK-2 cells exposed to high glucose and treated with Sal F. ResultCompared with the blank group， the model group showed decreased cell viability （P<0.01）， elevated levels LDH and IL-1β， increased proportion of PI-positive cells （P<0.01）， up-regulated protein levels of Bax， cytochrome C， Caspase-9， Caspase-3， and GSDME （P<0.01）， down-regulated protein level of Bcl-2 （P<0.01）， decreased mitochondrial membrane potential， and excessive ROS accumulation. Compared with the model group， Sal F repaired the high glucose-induced injury in HK-2 cells （P<0.05）， lowered the levels of LDH and IL-1β （P<0.05， P<0.01）， and decreased the proportion of PI-positive cells （P<0.01）. In addition， Sal F down-regulated the protein levels of Bax， cytochrome C， Caspase-9， Caspase-3， and GSDME and up-regulated the protein level of Bcl-2 （P<0.05， P<0.01）， increased the mitochondrial membrane potential， and decreased the accumulation of ROS in HK-2 cells. ConclusionSal F can reduce the production of ROS， restore the balance of mitochondrial membrane potential， and inhibit pyroptosis via the Bax/Caspase-3/GSDME signaling pathway to repair the high glucose-induced injury in HK-2 cells.

OBJECTIVE To study the effects of ginsenoside Rb 1（G-Rb1）on epithelial-mesenchymal transition （EMT）of renal tubular epithelial cells and its potential mechanism. METHODS The growth factor β1（TGF-β1）10 ng/mL was used to induce EMT of human renal tubular epithelial cells HK- 2. The morphological changes of HK- 2 cells were observed after treated with 10， 20，30 μmol/L G-Rb1 for 48 h. The transcriptional activities of biovector SBE in human embryonic kidney cell HEK 293 were determined after 24 h treatment with 1.0，2.5，5.0，10，20，30 μmol/L G-Rb1. Effects of above concentration of G-Rb 1 on the viability of HK- 2 cells were determined after 24 h of treatment. mRNA expressions of α-smooth muscle actin （α-SMA），collagen Ⅰ （COL-Ⅰ）and fibronectin （FN）in HK- 2 cells were detected after treated with 10，20，30 μmol/L G-Rb1 for 24 h. The expressions of α-SMA，Smad3，p-Smad3，COL-Ⅰ，FN and E-cadherin were detected after treated with 10，20，30 μmol/L G-Rb1 for 24 h. RESULTS G-Rb1 of 10-30 μmol/L significantly inhibited TGF-β1-induced EMT in HK- 2 cells and the increase of transcriptional activities of biovector SBE induced by TGF-β1（P＜0.05），but had no effects on relative activities of HK- 2 cells（P＞0.05）. The protein and mRNA expressions of α-SMA，COL-Ⅰ and FN ， the protein expressions of Smad 3 and p-Smad 3 were significantly up-regulated induced by TGF-β1（P＜0.05），while the protein expression of E-cadherin was significantly down- regulated（P＜0.05）；G-Rb1 could effectively reverse aboveprotein or mRNA expressions. CONCLUSIONS G-Rb1 can protect renal tubular epithelial cells from EMT induced byxiezhishen TGF-β1 to a certain extent ，which may be related to inhibiting the activation of TGF-β1/Smad3 signaling pathway.