Objective To evaluate the effect of dexmedetomidine on lipid peroxidation during lung ischemia-reperfusion (I/R) in rats.Methods Fifty-four pathogen-free healthy male Wistar rats,weighing 250-350 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (S group),I/R group,and dexmedetomidine group (Dex group).In group Dex,dexmedetomidine 5 μg/kg was injected via the caudal vein once a day for 2 consecutive days.The equal volume of normal saline was given in S and I/R groups.At 30 min after administration on 2nd day,lung I/R was produced by clamping the left hilum of lung for 45 min followed by 120 min of reperfusion in I/R and Dex groups.At 45 min of ischemia,and 60 and 120 min of reperfusion,6 rats selected from each group were sacrificed,and the left lungs were removed for examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),malondialdehyde (MDA) content and superoxide dismutase (SOD) activity.Results Compared with S group,the SOD activity was significantly decreased,and the MDA content and W/D ratio were significantly increased at 60 and 120 min of reperfusion in I/R and Dex groups (P＜0.05).Compared with I/R group,the SOD activity was significantly increased,and the MDA content and W/D ratio were significantly decreased at 60 and 120 min of reperfusion in Dex group (P＜0.05).The pathological changes of lungs were significantly attenuated in Dex group as compared with I/R group.Conclusion Dexmedetomidine mitigates lung I/R injury through inhibiting lipid peroxidation in rats.

Objective To investigate the effects and probable mechanism ofShenfu injection on the oxidaitve damage of H2O2-induced PC12 cells.Methods PC12 cells were cultured and exposed to 100μmol/L H2O2 for 1 h to establish the oxidative damage model. The protective effect ofShenfu injection was observed by the cell survival rate measured by colorimetric MTT assay, and the leakage rate of lactic dehydrogense (LDH). Western blot methods were used to detect the expression of NF-κB signaling pathway.Results Compared with the model group,Shenfu injection at 5, 10, 20 ml/L could improve the PC12 cells survival rate (83.11% ± 2.59 %, 87.99% ± 0.59%, 85.26% ± 1.07%vs. 73.82% ± 1.82%;P<0.01 orP<0.05), decrease the LDH leakage rate (32.75% ± 4.10%, 28.52% ± 1.14%, 35.79% ± 1.62%vs. 64.34% ± 3.18%;P<0.01 or P<0.05). Western blot results showed thatShenfu injection could protect the PC12 cells from oxidaitve damage by suppressing the p-p65/p65 (1.30 ± 0.10, 1.17 ± 0.06, 1.37 ± 0.15 vs. 1.70 ± 0.10;P<0.01 orP<0.05), p-IκBα/IκBα (1.07 ± 0.12, 1.00 ± 0.10, 1.03 ± 0.06 vs. 1.17 ± 0.06; P<0.01 orP<0.05).ConclusionShenfu injection has a obvious antioxidant effect on PC12 cells in vitro.

Aim To investigate the change of miR-124 expression in methamphetamine-induced addiction in PC12 cells and the possible regulatory mechanism that it involves.Methods PC12 cells were randomly divided into 6 groups as follows: control group, methamphetamine group, agomir Negative Control group, miR-124 agomir group, agomir Negative Control+methamphetamine group and miR-124 agomir+methamphetamine group.After the treatment, the total RNA and protein were extracted in PC12 cells.The expression of miR-124 was measured by Real-time PCR and the expression of GluR2 was determined by Western blot in PC12 cells.Results Compared with those in the control group, the expression of miR-124 was remarkably decreased and the expression of GluR2 was significantly increased in the methamphetamine group in PC12 cells.Compared with those in the agomir Negative Control+methamphetamine group, the expression of miR-124 was remarkably increased and the expression of GluR2 was significantly decreased in the miR-124 agomir+methamphetamine group in PC12 cells.Conclusion MiR-124 might involve in methamphetamine-induced addiction in PC12 cells by inhibiting GluR2.

Objective To evaluate the effect of lappaconitine on renal ischemia-reperfusion (I/R) injury in mice.Methods Thirty-six male Wistar rats,weighing 180-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and lappaconitine group (group LA).Renal I/R was induced by occlusion of the left renal pedicle for 45 min with atraumatic microclips followed by reperfusion,and the right kidney was removed after atraumatic microclips were released.At 30 min before reperfusion,lappaconitine 4 mg/kg was injected intraperitoneally in group LA,and normal saline 2 ml was given in S and I/R groups.In group S,the left renal pedicle was only isolated.At 5 and 24 h of reperfusion,blood samples were taken from the inferior vena cava for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations,and kidney specimens were obtained for histopathologic examination (with light microscope) and for determination of the expression of cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) in renal tissues (by immunohistochemistry).Results Compared with group S,the serum Cr and BUN concentrations were significantly increased,and the expression of COX-2 and MMP-2 in renal tissues was up-regulated at 5 and 24 h of reperfusion in I/R and LA groups.Compared with group I/R,the serum Cr and BUN concentrations were significantly decreased,the expression of COX-2 and MMP-2 in renal tissues was down-regulated at 5 and 24 h of reperfusion and histopathologic changes were reduced in group LA.Conclusion Lappaconitine can attenuate renal I/R injury through inhibiting the expression of COX-2 and MMP-2 in rats.

Objective To compare the therapeutical effect of puerarin, ligustrazine, ginsenoside Rb1, Hydroxysafflor yellow A on cerebral ischemia reperfusion mice. Methods The mice were randomly assigned for sham group, model group, puerarin group, ligustrazine group, ginsenoside Rb1 group, and Hydroxysafflor yellow A group, 24 mice for each group. All the groups were subjected to middle cerebral artery occlusion (MCAO) by 1 h ischemia and 24 h of reperfusion except the sham group. The puerarin, ligustrazine, ginsenoside Rb1, Hydroxysafflor yellow A were administrated by tail vein injection with 3μmol/kg at the onset of 1 h of ischemia. The neurologic deficit score, infarct area calculated by TTC staining, cerebral cortex blood flow monitored by laser doppler flowmetry, NO content measured by chemical colorimetry and western blot were applied to determine the expression for cleaved-caspase-3 and nuclear transcription factor NF-κB for each group. Results Compared with the model group, the infarct area (15.83%± 1.83%, 22.00%± 2.53%, 22.83%± 1.83%, 17.83%± 1.72%vs. 34.67%± 2.66%) in the puerarin group, ligustrazine group, ginsenoside Rb1 group, Hydroxysafflor yellow A group was significantly decreased (P<0.01 or P<0.05);the cerebral cortex blood flow (598.81 ± 9.90 μl/kg?min-1, 614.78 ± 9.20 μl/kg?min-1, 577.83 ± 5.55 μl/kg?min-1, 583.54 ± 7.98 μl/kg?min-1 vs. 548.43 ± 1.97 μl/kg?min-1) significantly increased (P<0.01 or P<0.05);the NO content (17.09 ± 1.18μmol/L, 18.54 ± 0.54μmol/L, 18.17 ± 0.49μmol/L, 15.10 ± 0.73μmol/L vs. 20.63 ± 0.73μmol/L) ignificantly decreased (P<0.01 or P<0.05);the expression of cleaved-caspase-3 (1.02 ± 0.08, 1.12 ± 0.04, 0.87 ± 0.08, 1.07 ± 0.08 vs. 1.30 ± 0.06) and NF-κB p-p65/NF-κB p65 (1.03 ± 0.19, 1.15 ± 0.05, 1.12 ± 0.08, 0.72 ± 0.08 vs. 1.45 ± 0.08) ignificantly decreased (P<0.01 or P<0.05) Conclusions Four Chinese herbal monomers could improve nerve and cerebral dysfunctions and ameliorate ischemia symptoms with varying degrees. The mechanisms were involved with the enhancement of cerebral cortex blood flow and inhibition of cell apoptosis and the activation of inflammatory signaling pathways.

Objective To evaluate the effects of sinomenine on the expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) during renal ischemia-reperfusion (I/ R) in rats.Methods Fifty-four male Wistar rats, weighing 180-220 g, were randomly divided into 3 groups (n=18 each) using a random number table: sham operation group (group S) , group I/R and sinomenine group (group SIN).Renal I/R was induced by clamping the left renal pedicle for 45 min followed by reperfusion in I/R and SIN groups.In group S, the bilateral renal pedicels were only exposed.Sinomenine 60 mg/kg was intraperitoneally injected at 30 min before reperfusion in group SIN, while the equal volume of normal saline was given in group S and group I/R.At 6, 12 and 24 h (T1-3) of reperfusion, 6 rats from each group were chosen, and blood samples were drawn from the hearts for determination of the serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The animals were then sacrificed, and the left kidney was removed and embedded into a paraffin block for determination of the expression of HIF-1α and VEGF in the renal tissues (by immuno-histochemistry).Results Compared with group S, the serum Cr and BUN concentrations were significantly increased at T1-3 , and the expression of HIF-1α was significantly up-regulated at T2,3 in group I/R and group SIN, and the expression of VEGF was significantly upregulated at T2,3in group I/R, and at T1-3 in group SIN.Compared with group I/R, the serum Cr concentration at T1-3 and serum BUN concentration at T2,3 were significantly decreased and the expression of HIF-1α at T2,3and VEGF at T1-3was significantly up-regulated in group SIN.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to up-regulation of the expression of HIF-1α and VEGF in the renal tissues of rats.

Objective To investigate the effect of sinomenine on the levels of nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-two healthy male Wistar rats, aged 6-9 weeks, weighing 180-220 g, were randomly assigned into 4 groups (n =18 each) using a random number table: control group (group C), sinomenine group (group SIN), group I/R, and sinomenine+I/R group (group SIN+I/R).The animals were anesthetized with 10％ chloral hydrate 350 mg/kg.The left renal pedicles were clamped with atraumatic microclips for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion to establish the model of renal I/R injury.Sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion in group SIN +I/R, and at the corresponding time point in group SIN.At 6, 8 and 12 h of reperfusion, the blood samples were drawn by cardiac puncture for measurement of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The left renal specimens were obtained for examination of pathological changes (with light microscopes) and for determination of the rate of NF-κB-positive cells and iNOS expression in renal tissues (by immunohistochemistry).Results Compared with group C, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly increased, and the expression of iNOS was up-regulated in I/R and SIN+I/R groups, and no significant change was found in the parameters mentioned above in group SIN.Compared with group I/R, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly decreased, and the expression of iNOS was down-regulated in group SIN+I/R.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN+I/R compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activity of NF-κB and down-regulated expression of iNOS in rats.

Objective To evaluate the effect of sinomenine on apoptosis in renal tubular epithelial cells of rats subjected to renal ischemia-reperfusion (I/R), and the relationship with C-Jun N-terminal kinase (JNK) signaling pathway.Methods Fifty-four male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were randomly divided into 3 groups (n =18 each) using a random number table: sham operation group (group S), I/R group and sinomenine group (group SIN).Renal ischemia was induced by occlusion of the left renal pedicle for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion in anesthetized rats in I/R and SIN groups.In group SIN, sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion, while the equal volume of normal saline was given instead of sinomenine at the same time point in S and I/R groups.Six animals in each group were selected at 0.5, 6 and 24 h of reperfusion, blood samples were collected by cardiac puncture for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.Immediately after blood sampling, the left kidney was removed for examination of pathological changes in renal tissues (with light microscopes) and for determination of phosphorylated JNK (p-JNK) and caspase-3 expression (by immune-histochemistry) and apoptosis in renal tubular epithelial cells (by TUNEL).The apoptotic rate was calculated.Results Compared with group S, the serum Cr and BUN concentrations were significantly increased, the expression of p-JNK and caspase-3 was up-regulated, and the apoptotic rate was increased in I/R and SIN groups.Compared with group I/R, the serum Cr and BUN concentrations were significantly decreased, the expression of p-JNK and caspase-3 was down-regulated, and the apoptotic rate was decreased in group SIN.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activation of p-JNK signaling pathway and reduced apoptosis in renal tubular epithelial cells of rats.

Objective To observe the effect of hydroxysafflor yellow A (HSYA) on the protection of blood brain barrier of cerebral ischemia mice, and explore the mechaniam. Methods Seventy-two C57/BL mice were divided into 6 groups: the sham operation group, the cerebral ischemia mice group, the TLR4 blocking group, the TLR4 blocking+cerebral ischemia mice group, the HSYA intervention+cerebral ischemia mice group, HSYA intervention+TLR4 blocking+cerebral ischemia mice group. Cerebral ischemia mice group were subjected to the middle cerebral artery occlusion (MCAO) model, TLR4 blocking was used, while TLR4 blocking was injected TLR4 antibody via right common carotid artery, and HSYA intervention group was injected 2 mg/kg HSYA by tail vein 0.5 h before cerebral ischemia. RT-PCR and western blot were applied to detect the mRNA and protein expression change of Wnt3a and β-catenin in each group. Results Compared with the cerebral ischemia mice group,the expression of TLR4 mRNA(1.63 ± 0.05,1.53 ± 0.04,1.84 ± 0.03 vs. 1.97 ± 0.05) significantly decreased (P<0.05 or P<0.01), the Wnt3a mRNA (0.56 ± 0.01, 0.58 ± 0.01, 0.50 ± 0.04 vs.0.42 ± 0.03),β-catenin mRNA(0.61 ± 0.03,0.74 ± 0.02,0.58 ± 0.04 vs.0.50 ± 0.03),Claudin-5 mRNA (0.54 ± 0.02, 0.58 ± 0.01, 0.47 ± 0.01 vs. 0.46 ± 0.02) mRNA significantly increased(P<0.05 or P<0.01), the protein expression of TLR4 (1.73 ± 0.05, 1.57 ± 0.03, 1.79 ± 0.08 vs. 1.89 ± 0.02) significantly decreased (P<0.05 or P<0.01), the protein expression of Wnt3a (0.67 ± 0.03, 0.74 ± 0.03, 0.57 ± 0.01 vs. 0.46 ± 0.01), Occludin(0.66 ± 0.02,0.73 ± 0.02,0.67 ± 0.01 vs.0.53 ± 0.01),Claudin-5(0.71 ± 0.01,0.73 ± 0.01,0.66 ± 0.01 vs. 0.64 ± 0.03) significantly increased (P<0.05 or P<0.01) in the TLR4 blocking+cerebral ischemia mice group, the HSYA intervention+cerebral ischemia mice group, HSYA intervention+TLR4 blocking+cerebral ischemia mice group. Conclusions TLR4 plays a critical regulatory role on the activation of Wnt3a and β-catenin in cerebral ischemic mice model. HSYA could intervene on the tight junction of cerebral ischemic brain through the intervention of Wnt3a and β-catenin, thus exerting the protection for cerebral ischemic brain.

Aim To investigate the effect of indirubin derivative PHⅡ-7 and TRAIL on proliferation in breast cancer cell MCF-7 and its MDR counterpart MCF-7/ADR and the mechanism.Methods Growth inhibition rate was examined respectively by MTT assay under treatment with TRAIL or PHⅡ-7 or in combination. Cell apoptosis and ROS production were examined by flow cytometry.The change of TRAIL receptors(DR4/DR5 )in mRNA was analysed by realtime PCR.Re-sults IC50 of PHⅡ-7 on MCF-7 and MCF-7/ADR was (4.49 ±1.55 ),(3.44 ±0.90 )μmol · L-1 respec-tively;MDA-MB-231 was TRAIL sensitive cell line, and apparently TRAIL induced apoptosis in MDA-MB-23 1 .Low concentration of PHⅡ-7 in combination with TRAIL could augment TRAIL-induced cytotoxic effect including apoptosis while TRAIL or PHⅡ-7 treatment alone had limited cytotoxity to those cells.Besides, PHⅡ-7 at this concentration had little toxicity to hu-man peripheral blood mononuclear cells even if in com-bination with TRAIL.PHⅡ-7 generated ROS produc-tion inside MCF-7 and MCF-7/ADR cells and up-regu-lated DR4/DR5 expression concentration dependently. Once upon ROS scavenger NAC involved,the effect of TRAIL receptors up-regualtion by expression was abro-gated.Conclusions PHⅡ-7 at low concentration could improve the sensitivities of breast cancer cell MCF-7 and MCF-7/ADR to TRAIL,the mechanism of which may be the ability of ROS production by PHⅡ-7 help up-regulated TRAIL receptor DR4,DR5 .Our re-search set a solid foundation for PHⅡ-7 in combination with TRAIL in future clinical application.