ABSTRACT
Objective:To investigate the effect of astragaloside Ⅳ（AST Ⅳ）and Notoginseng total saponins （NTS） combined with bone marrow mesenchymal stem cell （BMSC） transplantation on neural repair and angiogenesis in rats with cerebral ischemia. Method:The rats were randomly divided into a sham operation group， a model group， low- and high-dose AST Ⅳ + NTS groups， a BMSC infusion group， and low- and high-dose BMSC infusion+AST Ⅳ （10 and 20 mg·kg<sup>-1</sup>） + NTS group （25， 50 mg·kg<sup>-1</sup>）. BMSCs were isolated and purified by whole bone marrow adherent culture. The positive expression of surface markers of BMSCs （CD29， CD90， CD34， and CD45） was detected by flow cytometry. The focal cerebral ischemia model was established by middle cerebral artery occlusion （MCAO）. The PKH26-labeled BMSCs were injected into the tail vein of rats in the BMSC infusion group， once a day. The rats in the combination groups received BMSC injection once a day and intragastric administration of drugs twice a day. Other groups were administered twice a day by gavage. The sham operation group and the model group received the same amount of normal saline. Symptoms and signs of neurological deficits were assessed by the Longa method and the cerebral infarction rate was determined by TTC staining. The survival and vascularization ［double positive expression of PKH26/vascular endothelial growth factor （VEGF）］ after transplantation of BMSCs were observed by the immunofluorescence method. The protein expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was measured by Western blot. Result:BMSCs were properly isolated and cultured. The identification of surface markers CD29， CD90， CD34， and CD45 was consistent with the characteristics of BMSCs. The neurological deficit score and cerebral infarction rate of the model group were significantly increased （<italic>P</italic><0.01）. All drugs and cell transplantation could alleviate the above pathological changes in varying degrees. The strongest effect was observed in high-dose BMSC infusion+AST Ⅳ+NTS group （<italic>P</italic><0.01）， which was superior to those in the AST Ⅳ+NTS groups or the BMSC infusion group. BMSC injection helped cells survive in the ischemic brain tissues and promoted angiogenesis， and this effect could be enhanced by the combination with drugs. After cerebral ischemia， the expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was increased， and the effect in the BMSC infusion+AST Ⅳ+NTS groups was the strongest （<italic>P</italic><0.01）. Conclusion:AST Ⅳ combined with NTS can promote the survival of transplanted BMSCs and facilitate angiogenesis after target repair of damaged blood vessels after cerebral ischemia. The mechanism may be related to the improvement of the local microenvironment in the brain after cerebral ischemia and the promotion of the survival and differentiation of transplanted stem cells.
ABSTRACT
The present study was aimed to investigate the protective effect and anti-inflammation mechanism of astragaloside IV (AST-IV) on cerebral ischemia and reperfusion injury. Following the establishment of cerebral ischemia and reperfusion model in rats by modified suture method, neurological deficit scores and cerebral infarct volume were used to evaluate the pharmacological effect of AST-IV against cerebral ischemia-reperfusion injury. Western blot was used to detect the expression levels of NLRP3, pro-Caspase-1, Caspase-1, pro-IL-1β, IL-1β, pro-IL-18, IL-18, phosphorylated and total nuclear factor kappa B (NF-κB)/p65 protein in the brain tissue. The results showed that compared with model group, the intervention of AST-IV decreased the neurological deficit scores, reduced the cerebral infarct volume, decreased the levels of NLRP3, Caspase-1, pro-IL-1β, IL-1β, pro-IL-18 and IL-18, and inhibited the expression of phosphorylated NF-κB in brain tissue. The results suggest that AST-IV has a protective effect against cerebral ischemia and reperfusion injury, and its mechanism is related to inhibiting the phosphorylation of NF-κB and NLRP3 inflammasome activation.
Subject(s)
Animals , Rats , Brain Ischemia , Drug Therapy , Infarction, Middle Cerebral Artery , Drug Therapy , Inflammasomes , Metabolism , NF-kappa B , Metabolism , NLR Family, Pyrin Domain-Containing 3 Protein , Metabolism , Phosphorylation , Rats, Sprague-Dawley , Reperfusion Injury , Drug Therapy , Saponins , Pharmacology , Triterpenes , PharmacologyABSTRACT
Pyroptosis is a form of inflammatory programmed cell death activated by caspase-1 and caspase-4/5/11, and involves in the pathogenesis of infectious diseases and nervous system diseases. Pyroptosis is mediated by canonical inflammasome pathway and non-canonical inflammasome pathway. The canonical inflammasome pathway is activated in stroke and aggravates brain injury. Inhibition of inflammasome, caspase-1, IL-1β and IL-18 ameliorates brain injury. These studies indicate that canonical inflammasome pathway contributes to post-stroke brain injury, therefore, pyroptosis has become a potential therapeutic target for preventing excessive cell death during stroke. We reviewed the relationship between pyroptosis and stroke to provide some perspectives on future researches in this field.
ABSTRACT
The aim is to study the effect of astragaloside Ⅳ (AST Ⅳ) combined with Panax notoginseng saponins (PNS) on cerebral ischemia-reperfusion injury, and to probe the synergistic mechanism through the pharmacokinetics of the four major components such as AST Ⅳ, ginsenoside Rg₁ (Rg₁), ginsenoside Rb₁ (Rb₁), notoginsenoside R₁ (R₁) in cerebral ischemia-reperfusion rats. Following the establishment of cerebral ischemia/reperfusion model in rats by modified suture method, neurological function score, cerebral infarction area and pathomorphology were used to evaluate the pharmacological effect that the combination of AST Ⅳ and PNS antagonized cerebral ischemia-reperfusion injury; the contents of AST Ⅳ, Rg₁, Rb₁, R₁ in rat plasma of different time points were determined with ultra performance liquid chromatography tandem massspectrometry (UPLC-MS/MS), pharmacokinetic parameters were calculated and pharmacokinetics changes of the main effective components were analyzed. The results showed that AST Ⅳ, PNS alone and their combination could reduce the cerebral infarction area of rats, relieve the behavioral scores of neurologic deficit, improve the pathological changes after cerebral ischemia, the effects of the combination were better. Among AST Ⅳ, Rg₁, Rb₁, R₁, the area under the curve (AUC) was significantly increased, the mean residence time of (MRT0-t) was delayed, the peak concentration (Cmax) was significantly raised, the apparent volume of distribution (Vz/F) was reduced, and the clearance rate in vivo was significantly slowed. It suggested that AST Ⅳ combined with PNS has synergistic enhancement on anti-cerebral ischemia/reperfusion injury, moreover, make the pharmacokinetic behavior of the main effective components change, the mechanism may be associated with prolonging the retention time of the effective components in cerebral ischemia condition, elevating the bioavailability.
ABSTRACT
The aim is to study the effect and its mechanism of Astragalus Radix combined with Angelicae Sinensis Radix on the proliferation of hematopoietic stem cells(HSCs) in senescence model. After drug-containing plasma of rats was prepared via intragastric administration, HSCs of mice were cultured in vitro, and then they were divided into blank control group, model group, blank plasma group, Astragalus Radix + Angelicae Sinensis Radix 1∶1 group and 10∶1 group, Angelicae Sinensis Radix plasma group, and Astragalus Radix plasma group. HSCs senescence model was induced by using tert-butyl hydrogen peroxide(t-BHP), and intervened by drug-containing plasma. Cells senescence rate was tested by SA-β-galactosidase staining method; cell cycle distribution was determined by flow cytometry; Cyclin D1, P21, and P53 mRNA were measured with RT-PCR, and Cyclin D1 protein expression was measured by Western blot. Results showed that after being induced by t-BHP, senescence rate of HSCs was increased; cell proliferation ability was decreased; count of G₀/G₁ phase cells was increased; count of G₂/M+S phase cells was reduced; Cyclin D1 expression was down-regulated while P53, P21 expression was up-regulated, which were reversed by Astragalus Radix + Angelicae Sinensis Radix 1∶1 and 10∶1, single Angelicae Sinensis Radix, and single Astragalus Radix plasma. Furthermore, the above effects were most obvious in Astragalus Radix+Angelicae Sinensis Radix 1∶1 group. These results suggested that t-BHP can promote HSCs senescence and reduce cell proliferation ability. Angelicae Sinensis Radix, Astragalus Radix and their combinations can inhibit HSCs senescence, promote HSCs proliferation as well as cell cycle conversion; moreover, the effects of 1∶1 Astragalus Radix+Angelicae Sinensis Radix were strongest. The mechanisms may be related to up-regulating the expression of cell cycle positive regulator, down-regulating the expression of cell cycle negative regulator, thus promoting the cells to enter the proliferation phase from the stationary phase.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effects and molecular mechanisms of the combination between total Astragalus extract (TAE) and total Panax notoginseng saponins (TPNS) against cerebral ischemia-reperfusion injury.</p><p><b>METHODS</b>C57BL/6 mice were randomly divided into sham-operated group, model group, TAE (110 mg/kg) group, TPNS (115 mg/kg) group, TAE-TPNS combination group and Edaravone (4 mg/kg) group, treated for 4 days, then, cerebral ischemia-reperfusion injury was established by bilateral common carotid artery (CCA) ligation for 20 min followed by reperfusion for 1 and 24 h.</p><p><b>RESULTS</b>TPNS could increase adenosine triphosphate (ATP) level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate (ADP) contents and Na-K-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinase1/2 (p-JNK1/2), cytochrome C (Cyt C), cysteine aspartic acid-specific protease (Caspase)-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone.</p><p><b>CONCLUSION</b>The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.</p>
ABSTRACT
This paper was aimed to explore the effects of glycosides, the effective component of Buyang Huanwu decoction, and its main active components such as astragaloside Ⅳ, amygdalin, peoniflorin and their combinations on vascular smooth muscle cells (VSMC) proliferation, clarify the major active materials of anti-VSMC proliferation and investigate the mechanisms via the signal transduction pathway. Plasma containing drug was prepared via oral administration in rats. VSMCs of rats aorta were cultured, and then VSMC proliferation was stimulated by using platelet derived growth factor (PDGF).The plasma containing drug was added to detect the activity of cell proliferation, cell cycle and related protein expressions of signaling pathway such as extracellular signal-regulated kinase (ERK), phos-phatidylinositol-3-kinase/protein kinase B (PI3K/Akt) and Janus kinase/signal transducer and activator of transcription (JAK/STAT). After being stimulated by PDGF, the proliferation activity of VSMC was strengthened (P<0.01), G₀/G₁ phase cells were decreased (P<0.01), S/M phase cells were increased (P<0.01), and PcNA, cyclin D1 protein expressions related to cell cycle were up-regulated (P<0.01). Glycosides, astragaloside Ⅳ, amygdalin, peoniflorin and their combinations could inhibit the cell proliferation (P<0.05 or P<0.01) in a dose-effect relationship and time-effect relationship. They could increase G₀/G₁ phase cells (P<0.01), decrease S/M phase cells (P<0.01), and down-regulate the protein expressions of PCNA, cyclin D1 (P<0.01); and the effects of the combinations were greater than those of single active component (P<0.05). After VSMC proliferation was induced by PDGF, p-ERK1/2 expression was increased (P<0.01), PI3K expression was down-regulated while p-PI3K expression was up-regulated (all P<0.01), and STAT3expression was reduced while p-STAT3 expression was increased (all P<0.01). Glycosides, astragaloside Ⅳ, amygdalin, peoniflorin and the combinations of these active components could reduce p-ERK1/2 expression (P<0.05), increase PI3K expression (P<0.01), decreasep-PI3K expression (P<0.05 or P<0.01), increase STAT3 expression (P<0.01), and decrease p-STAT3 expression (P<0.05 or P<0.01). These results suggested that PDGF could induce the cell cycle conversion of VSMC, leading to VSMC proliferation. The mechanism was related to the activation of ERK, PI3K/Akt and JAK/STAT signaling pathways. Glycosides and its main active components such as astragaloside Ⅳ, amygdalin, peoniflorin and their combinations can inhibit the cell cycle conversion of VSMC, with the effect against VSMC proliferation, and the mechanisms may be associated with the inhibition of PI3K/Akt, mitogen-activated protein kinase (MAPK) and JAK/STAT signaling pathways. astragaloside Ⅳ, amygdalin and peoniflorin were the major active materials of anti-VSMC proliferation, and their combination showed enhanced effect.
ABSTRACT
Autophagy is a lysosome-mediated degradation process for non-essential or damaged cellular constituents, playing an important homeostatic role in cell survival, differentiation and development to maintain homeostasis. Autophagy is involved in tumors as well as neurodegenerative, cardiovascular and cerebrovascular diseases. Recently, active compounds from traditional Chinese medicine (TCM) have been found to modulate the levels of autophagy in tumor cells, nerve cells, myocardial cells and endothelial cells. Ischemic stroke is a major cause of neurological disability and places a heavy burden on family and society. Regaining function can significantly reduce dependence and improve the quality of life of stroke survivors. In healthy cells, autophagy plays a key role in adapting to nutritional deprivation and eliminating aggregated proteins, however inappropriate activation of autophagy may lead to cell death in cerebral ischemia. This paper reviews the process and the molecular basis of autophagy, as well as its roles in cerebral ischemia and the roles of TCM in modulating its activity.
Subject(s)
Humans , Autophagy , Brain Ischemia , Pathology , Medicine, Chinese Traditional , Reperfusion Injury , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To compare effects of different point combinations of Zusanli (ST 36) for improving acute gastric mucosa injury and study on the mechanism.</p><p><b>METHODS</b>One hundred rats were randomly divided into 10 groups, Zusanli (ST 36) group (group A); Zusanli and Neiguan (PC 6) group (group B); Zusanli and Zhongwan (CV 12) group (group C); Zusanli and Gongsun (SP 4) group (group D); Zusanli, Neiguan and Zhongwan group (group E); Zusanli, Neiguan and Gongsun group (group F); Zusanli, Zhongwan and Gongsun group (group G); Zusanli and Neiguan, Zhongwan, Gongsun group (group H); model group (group I); blank control group (group J), 10 rats in each group. Gastric mucosa injury model was made by intragastric infusion of dehydrated alcohol (0. 6 mL/100 g). The gastric mucosa injury index (UI), epidermal growth factor (EGF), nitric oxide (NO) and gastrin (GAS) contents were detected.</p><p><b>RESULTS</b>Contents of EGF and NO were significantly increased and GAS content decreased in all of the EA groups as compared with those in the model group (P<0. 01 or P<0. 05), with no significant differences among group A, B and D, and significant differences as group A compared with group C, F and group H compared with other EA groups.</p><p><b>CONCLUSION</b>Different point combinations of Zusanli (ST 36) can improve acute gastric mucosa injury, with the strongest effect in the Zusanli and Neiguan, Zhongwan, Gongsun group.</p>
Subject(s)
Animals , Female , Male , Rats , Acupuncture Points , Acute Disease , Epidermal Growth Factor , Blood , Ethanol , Toxicity , Gastric Mucosa , Pathology , Gastrins , Blood , Nitric Oxide , Blood , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To establish the mathematical kinetic model of the components extracted from the FTMC (formulae of the traditional Chinese medicine) and analyze parameters of the astragaloside IV extracted from the BYHWD (Buyang Huanwu decoction).</p><p><b>METHOD</b>The model, including algebra and differential groups, have been set up according to the FICK discipline and Noyes-whitney soluted theories, as well as two transfer diffusive processes ((1) from protoplasate to apoplasmic, also from material compartment interior cell membrane to outside compartment; (2) apoplasmic to solution, also from outside compartment to solvent compartment) on components extraction from the FTMC. The equation groups, according to laplace transform, have been given a expression as solutions, which indicate the quantitative changes of the component concentration in solvent vs. time. The model kinetic parameters have been analyzed, meanwhile the parameters of the astragaloside IV in the BYHWD under 100 degrees C, extracted by water, have been analyzed by way of this model:</p><p><b>RESULT</b>It has been established a mathematical model that consists of three parts of e exponent. The kinetic parameters: M, alpha, N, beta, L, pi, K, k1', k2', rho1, rho2, tmax, Cmax, AUC, w0, P, D of the BYHWD were respectivelly 0.061 27% , 0.280 2 min(-1), - 1.027% , 0.008 965 min(-1), 1.077%, 0.002 665 min(-1), 3.451 x 10(-3) min(-1), 3.188 x 10(-3) min(-1), 0.375 9 min(-1), 1.420 min, 0.754 7 min, 184.9 min, 0. 0572 1 mg x mL(-1), 289.9 min, 0.070 11%, 46.24%, 22. 35%.</p><p><b>CONCLUSION</b>The kinetic model, applied to isolated system, can have been of the rule of multiplex linear. Each parameters can be analyzed completely.</p>
Subject(s)
Algorithms , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Kinetics , Mathematics , Models, Biological , Plants, Medicinal , Chemistry , Saponins , Chemistry , Triterpenes , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Sanhuang Jiangtang recipe (SJR) on renin-angiotensin system in local myocardium in diabetic rats.</p><p><b>METHODS</b>Rats were made into diabetes model by intraperitoneally administering of streptozocin, and medicated through gastrogavage with SJR (50 g/kg), gliclazide (20 mg/kg), captopril (15 mg/kg) and nitrendipine (30 mg/kg) respectively, for successive 8 weeks, started from 2 weeks after modeling. Levels of fasting blood sugar (FBS), serum insulin (Ins), heart/body weight ratio (H/BW), myocardial angiotensin II (Ang II), angiotensin converting enzyme (ACE) and aldosterone (ALD) were determined. And the mRNA expression of type I angiotensin receptor (AT1R) in myocardium were detected by RT-PCR assay.</p><p><b>RESULTS</b>As compared with those in the normal rats, levels of FBS, H/BW, Ang II, ACE, ALD and AT1R mRNA expression were higher (all P < 0.05) and level of serum Ins was lower (P < 0.01) in the model rats. SJR, gliclazide, captopril and nitrendipine could slightly reduce the blood sugar level in model rats, but with no increase of serum Ins. All the four drugs could reduce H/BW, Ang II, ACE and AT1R mRNA expression. SJR and captopril could also decrease the ALD content in myocardium.</p><p><b>CONCLUSION</b>Cardiac hypertrophy has been induced in 10 weeks after diabetic modeling. Activation of local myocardial RAS is related to the genesis of diabetic cardiomyopathy. SJR, gliclazide, captopril and nitrendipine could antagonize the genesis of diabetic cardiomyopathy, the mechanism is related to the inhibition of RAS activation in local myocardium.</p>
Subject(s)
Animals , Male , Rats , Cardiomegaly , Diabetes Mellitus, Experimental , Metabolism , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Hypoglycemic Agents , Pharmacology , Therapeutic Uses , Myocardium , Metabolism , Pathology , Rats, Sprague-Dawley , Renin-Angiotensin SystemABSTRACT
<p><b>OBJECTIVE</b>To study the effects of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion.</p><p><b>METHOD</b>Cerebral ischemia model in gerbils was produced by ligating bilateral common carotid artery. The dynamic expressin of GFAP were determined by immunochemistry after cerebyal ischemia for 15 min followed by reperfusion for 24 hours and 48 hours.</p><p><b>RESULT</b>GFAP positive reactions reached a peak after cerebral ischemia for 15 min followed by reperfusion for 24 hours. Bu Yang Huan Wu Decoction decreased the expression. GFAP positive reactions decreased after cerebral ischemia for 15 min followed by reperfusion for 48 hours, Bu Yang Huan Wu Decoction increased the expression.</p><p><b>CONCLUSION</b>The regulation of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion may be related to repairing process after cerebral ischemia.</p>
Subject(s)
Animals , Female , Male , Astrocytes , Brain Ischemia , Drugs, Chinese Herbal , Pharmacology , Glial Fibrillary Acidic Protein , Metabolism , Hippocampus , Metabolism , Plants, Medicinal , Chemistry , Reperfusion Injury , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To compare the effects of Buyang Huanwu Decoction (BYHWD) and its active regions combination on brain heat shock protein 70 (HSP70) expression.</p><p><b>METHODS</b>Cerebral ischemia model gerbils produced by ligating bilateral common carotid artery were injected intraperitoneally by BYHWD and its active regions combination separately. HSP70 mRNA expression was determined by Northern blot test and protein expression determined by Western blot test 48 hrs after ischemia for 15 min and reperfusion.</p><p><b>RESULTS</b>The level of HSP70 mRNA and protein expression in ischemic/reperfusion brain tissue increased significantly. BYHWD and its active regions combination could decrease the HSP70 mRNA expression obviously, the inhibition effect of BYHWD was more significant than that of its active regions combination, but neither of them showed any effect on HSP70 protein expression.</p><p><b>CONCLUSION</b>BYHWD and its active regions combination might down-regulate HSP70 mRNA expression in ischemic brain tissue through its anti-ischemic effect.</p>