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The aim of this study was to explore the effect by which the combination of Astragaloside IV (AST IV) and Ginsenoside Rg1 (Rg1) resisted autophagic injury in PC12 cells induced by oxygen glucose deprivation/reoxygenation (OGD/R). We studied the nature of the interaction between AST IV and Rg1 that inhibited autophagy through the Isobologram method, and investigated the synergistic mechanism via the PI3K I/Akt/mTOR and PI3K III/Becline-1/Bcl-2 signaling pathways. Our results showed that, based on the 50% inhibiting concentration (IC50), AST IV combined with Rg1 at a 1:1 ratio resulted in a synergistic effect, whereas the combination of the two had an antagonistic effect on autophagy at ratios of 1:2 and 2:1. Meanwhile, AST IV and Rg1 alone increased cell survival and decreased lactate dehydrogenase (LDH) leakage induced by OGD/R, reduced autophagosomes and the LC3 II positive patch, down-regulated the LC3 II/LC3 I ratio and up-regulated the p62 protein; the 1:1 combination enhanced these effects. Mechanistic study showed that Rg1 and the 1:1 combination increased the phosphorylation of PI3K I, Akt and mTOR; the effects of the combination were greater than those of the drugs alone. AST IV and the 1:1 combination suppressed the expression of PI3K III and Becline-1, and the combination elevated Bcl-2 protein expression; the effects of the combination were better than those of the drugs alone. These results suggest that after 2 h-OGD followed by reoxygenation for 24h, PC12 cells suffer excessive autophagy and damage, which are blocked by AST IV or Rg1; moreover, the combination of AST IV and Rg1 at a 1:1 ratio of their IC50 concentrations has a synergistic inhibition on autophagic injury. The synergistic mechanism may be associated with the PI3K I/Akt/mTOR and PI3K III/Becline-1/Bcl-2 signaling pathways.

The aim of this study was to investigate the pharmacokinetics and pharmacodynamics of seven main active components of Mahuang decoction (MHD) and its time-concentration-effect relationship. The asthmatic rat model was established by the method of ovalbumin (OVA) sensttization. The plasma concentrations of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamic acid, glycyrrhizic acid in asthmatic model rat were investigated by a selective and rapid HPLC/MS-MS method. Simultaneously, the asthma-involved cytokines including leukotrienes B4 (LTB4), thromboxane B2 (TXB2), 6-Keto-Prostaglandin F1α (6-K-PGF1α) and histamine (HIS) levels in rat plasma were determined by using ELISA. A mathematics method was applied to assess the trend of percentage rate of change among different time intervals of the seven components. The sigmoid E max function was used to establish the PK-PD modeling of MHD. The results indicated that MHD could control or ameliorate asthma. There was a hysteresis between the peaked drug concentration and maximum therapeutic effect of MHD. The PK-PD curves of MHD showed clockwise or counter-clockwise hysteresis loop. In addition, amygdalin might exert a more significant influence on regulating cytokines levels in asthmatic rats among the seven components of MHD.

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.

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.

To explore the effects of alkaloids and glycosides extracted from Buyang Huanwu Decoction (BYHWD), a compound of traditional Chinese herbal medicine, on aortic intimal hyperplasia and the expression of the proliferating cell nuclear antigen (PCNA) in rats with aortic intimal injuries.The vessel restenosis model was established by denuding aortic endothelium with domestic balloon catheter in rats. Drugs were administered intragastrically on the first day after operation. The injured segments of aorta were taken on the fifteenth day after operation to determine the degree of intimal hyperplasia and observe the expression of PCNA.Aortic intimal hyperplasias were very obvious on the fifteenth day after operation. The media hyperplasias in the drug-treated groups were not significant (P0.05), but the intimal hyperplasia were remarkable as compared with that in the sham-operated group (P0.01). The degrees of intimal hyperplasia in BYHWD, alkaloids, glycosides and atorvastatin groups were less than that in the untreated group (P0.01). There was significant difference of PCNA expression between the untreated group and the sham-operated group (P0.01). The expressions of PCNA in alkaloids, glycosides, and atorvastatin groups were higher than that in the sham-operated group (P0.01, P0.05), but still lower than that in the untreated group (P0.01). The expression of PCNA in BYHWD group was higher than that in the sham-operated group (P0.01), and no significant difference was found between the BYHWD group and the untreated group (P0.05).Alkaloids and glycosides extracted from BYHWD can inhibit intimal hyperplasia induced by denuding arterial endothelium with domestic balloon catheter in rats. Alkaloids and glycoside may be among basal substances in BYHWD inhibiting intimal hyperplasia of blood vessel, the effect of which may relate to down-regulating the expression of PCNA.

To investigate the effects of Panax notoginseng saponins (PNS) on mRNA expressions of interleukin-1 beta (IL-1 beta), interleukin-1 receptor type I (IL-1RI), interleukin-1 receptor antagonist (IL-1ra), intercellular adhesion molecule-1 (ICAM-1), cysteinyl-aspartate specific protease-1 (caspase-1), caspase-3 and caspase-8 after cerebral ischemia-reperfusion in rats.Focal cerebral ischemia reperfusion in rats was induced by the method of nylon monofilament via the internal carotid artery. PNS was administered intraperitoneally respectively five minutes before cerebral ischemia and twelve hours after cerebral ischemia. After cerebral ischemia for two hours followed by reperfusion for twenty two hours, the mRNA expressions of IL-1 beta, IL-1RI, IL-1ra, ICAM-1, caspase-1, caspase-3 and caspase-8 in brain tissue were determined by reverse transcription polymerase chain reaction assay.After cerebral ischemia for two hours followed by reperfusion for twenty two hours, the mRNA expression levels of IL-1 beta, IL-1RI, IL-1ra, ICAM-1, caspase-1, caspase-3 and caspase-8 in brain tissue in the untreated group were obviously elevated as compared to those in the sham-operation group (P0.05 or P0.01). The mRNA expression levels of IL-1 beta, IL-1RI, IL-1ra in brain tissue in the PNS group were lower than those in the untreated group, but higher than those in the sham-operation group, and without statistical differences as compared to those in the sham-operation group and in the untreated group (P0.05). The mRNA expression level of caspase-3 in brain tissue in the PNS group was significantly lower than that in the untreated group (P0.05), but PNS had no effect on the mRNA expression levels of ICAM-1, caspase-1 and caspase-8 in brain tissue.PNS can inhibit the mRNA expression of caspase-3, slightly inhibit the mRNA expressions of IL-1 beta and its correlative inflammatory factors in brain tissue. The protective effects of PNS on cerebral injury induced by ischemia-reperfusion may be related to inhibiting the mRNA expressions of caspase-3, IL-1 beta and its correlative inflammatory factors in brain tissue.

Aim of the study: the effect of total saponins of “panax notoginseng root” on aortic intimal hyperplasia and the expressions of cell cycle protein and extracellular matrix in rats Materials and methods Sprague-Dawley rats were randomly divided into sham-operated, control, TSPN and atorvastatin group. Rat aorta intima in all groups were injured by insertion of domestic balloon catheter into the aortae except sham-operated rats. Drugs were administrated orally from the second day after vascular injury and continued for 14 days. The injured segments of aortae were collected on the sixteenth day after operation to observe the morphological changes of vascular structure and to examine the expressions of proliferating cell nuclear antigen(PCNA), cyclinD1, cyclinE, collagen I(Col-I), fibronect(FN), matrix metalloproteinase-9(MMP-9) and tissue inhibitor metalloproteinase-1(TIMP-1). Results: TPNS significantly inhibited the vascular intimal hyperplasia. TPNS significantly lowered the expression of PCNA, cyclinE, cyclinD1, FN and MMP-9. TPNS had no significant impacts on the expression of Col-I and TIMP-1. Conclusions: Our studies indicated that TSPN could inhibit vessel restenosis after vascular intimal injury, and its mechanisms may be related to the blockage of the excessive proliferation of VSMC, the reduction of ECM protein deposition in the endometrium, and the degradation of ECM protein.

Aim of the study The inhibitive effect of BuYang HuanWu Decoction (BYHWD) and its major components on vascular intimal hyperplasia and the expressions of cell cycle protein and extracellular matrix protein. Materials and methods Sprague-Dawley rats were randomly divided into sham-operated, control, alkaloid, glycoside, BYHWD and atorvastatin groups. Rat aorta intima in all groups were injured by insesion of domestic balloon catheter into the aortae except sham-operated rats. Drugs were administrated orally from the second day after vascular injury and continued for 14 days. The injured segments of aortae were collected on the sixteenth day after operation to observe the morphological changes of vascular structure and to examine the expressions of proteins in vascular cells associated with cell cycle including proliferating cell nuclear antigen(PCNA), cyclinD1 and cyclinE, and extracellular matrix(ECM) proteins including collagen I (Col-I) and fibronectin (FN), further to discover the involved biologically active substances and the potential mechanisms. Results Alkaloid and glycosid isolated from BYHWD were more effective than BYHWD in the inhibition of intimal hyperplasia and the expressions of PCNA, cyclinD1, cyclinE, Col-I and FN, suggesting that alkaloid and glycoside may be the main components of BYHWD responsible for the observed inhibition of excessive hyperplasia of vascular intima. Conclusions The mechanism associated with the anti-hyperplasia activity of BYHWD and its effective components may be related to the blockage of cell cycles of VSMC, and the inhibition of the ECM protein synthesis, even the increased degradation of ECM proteins.

To investigate effect of the four kinds of active fractions extracted from Buyang Huanwu Decoction (BYHWD) on caspase expression in rats after focal cerebral ischemic reperfusion.The focal cerebral ischemia/reperfusion model rats were established by middle cerebral artery occlusion for 2 hrs followed with reperfusion for 46 hrs. Before and at 12th, 24th and 36th hour after cerebral ischemia, the rats were administered with alkaloid, glycoside, polysaccharide and aglycone from BYHWD respectively, and nimodipine was given as control medicine to observe the effect of them on protein expression of caspase-1, caspase-3 and caspase-8 using immunohistochemical method.Protein expression of caspase-1 and caspase-3 increased in the injured lateral brain tissue after cerebral ischemia/reperfusion. Caspase-1 expression were observed mainly in choroids, while caspase-3 expression in hippocampus, cortex and medulla. All the four kinds of active fractions from BYHWD could inhibit caspase-1 expression, while nimodipine couldn't. Caspase-3 expression in hippocampus and medulla could be inhibited by alkaloid, that in hippocampus, cortex and medulla could be inhibited by glycoside and aglycone, and that in hippocampus and medulla by nimodipine, however, polysaccharide showd no effect on it. As for caspase-8 expression, no effect on it was shown in all groups.Alkaloid, glycoside, polysaccharide and aglycone from BYHWD could relieve the inflammatory reaction occurred after cerebral ischemia/reperfusion by inhibiting caspase-1 expression to decrease production of inflammatory cytokine. Alkaloid, glycoside and aglycone could reduce neuronal apoptosis by inhibiting caspase-3 expression to antagonize the delayed neuronal death after cerebral ischemia. The 4 kinds of active fractions may be the main material basis for BYHWD in preventing ischemia/reperfusion cerebral injury.