Objective:To investigate the effects of Xingnao Yisui Decoction on the behaviour of vascular dementia (VaD) rats and the apoptosis of neurons in the CA1 region of the hippocampus; To explore its mechanism of action.Methods:Totally 60 adult SD rats with similar levels of cognitive function were selected and divided into Xingnao Yisui Decoction high-, medium- and low-dosage groups, donepezil group, model group and sham-operation group according to the random number table method, with 10 rats in each group. Except for the sham-operation group, the VaD model was prepared using a modified bilateral common carotid artery ligation method in all the groups. Xingnao Yisui Decoction high-, medium- and low-dosage groups were gavaged with 19.62, 9.81, 4.91 g/(kg·d) of Xingnao Yisui Decoction respectively, the donepezil group was gavaged with 0.5 mg/(kg·d) of donepezil solution, and the sham-operation group and the model group were given equal amount of saline gavage, 1 time/d for 4 weeks. Morris water maze experiment was used to observe the behavioral changes in each group, TUNEL staining was used to observe the apoptosis of neurons in CA1 area of hippocampus. Realtime PCR was used to detect the mRNA expression of NF-κB and MMP-9. ELISA was used to detect the changes of serum NF-κB, MMP-9, p53, Bcl-2 and Bax protein levels.Results:Compared with the model group, the escape latency of rats in the donepezil group and Xingnao Yisui Decoction high-, medium- and low-dosage groups was shortened ( P<0.05); the times of crossing the platform and the stay time in the target quadrant increased ( P<0.05); the number of apoptotic cells in hippocampal CA1 region decreased ( P<0.01); the number of apoptotic cells in hippocampal CA1 region decreased ( P<0.01); the mRNA expression of NF- κB and MMP-9 in hippocampal mRNA level decreased ( P<0.01); the content of NF-κB, MMP-9, p53 and Bax decreased ( P<0.01), while the content of Bcl-2 increased ( P<0.01). Conclusion:Xingnao Yisui Decoction can effectively mediate the NF-κB-MMP-9 pathway to improve the permeability of the blood-brain barrier and regulate p53-Bcl-2-Bax to reduce apoptosis of neurons in CA1 area of hippocampus, thus significantly improve the learning and memory ability of VaD rats.

AIM:To investigate the effect of activation of retinoid X receptor (RXR) on transforming growth factor β1 (TGF-β1) induced collagen synthesis under hypoxic environment in rat cardiac fibroblasts (CFs) and underlying molecular mechanisms .METHODS: CFs were cultured using myocardial tissue with dry method .Hypoxic environment was established for CFs by continuous nitrogen supplement .Type I and type III collagens in supernatants were detected by ELISA.Nuclear and cytoplasmic extractions were prepared using NE-PER nuclear and cytoplasmic extraction reagents .The protein levels of Smad2 and p-Smad2 were determined by Western blot and immunocytochemical staining .RESULTS:Un-der hypoxic condition , TGF-β1 (0.01~10 μg/L) increased the synthesis of type I and type III collagens in a dose-de-pendent manner in the CFs .At the concentration of 5μg/L, the synthesis of collagen I and III was significantly increased as compared with control group (P<0.01).RXR agonist 9-cis-retinoic acid (9-cis-RA;10 -9 ~10 -6 mol/L) decreased TGF-β1 (5μg/L)-induced synthesis of type I and III collagens in a dose-dependent manner in the CFs under hypoxic con-dition.The synthesis of type I and type III collagens was significantly inhibited by 9-cis-RA (P<0.01).Smad2 inhibitor ( 20 nmol/L) showed similar inhibitory effect on the synthesis of type I and III collagens induced by TGF -β1 under hypoxic condition.Compared with TGF-β1 intervention group, the cytoplasmic level of p-Smad2 in the CFs was significantly in-creased in TGF-β1+9-cis-RA group, but the nuclear p-Smad2 level was significantly decreased (P<0.05).CONCLU-SION:Retinoid X receptor agonist 9-cis-RA inhibits TGF-β1-induced synthesis of type I and type III collagens in the CFs by repressing p-Smad2 nuclear translocation under hypoxic condition .

[ABSTRACT]AIM:ToexploretheeffectofretinoidXreceptor(RXR)agonistbexarotene(Bex)andvitaminD receptor (VDR) agonist calcitriol (Cal) on the expression of nuclear factor-kappa B (NF-κB) and the development of atherosclerosis in streptozotocin-induced diabetic apolipoprotein E knockout ( STZ-ApoE-/-) mice.METHODS: Male mice were treated for 12 weeks as follows:(1) C57+vehicle;(2) ApoE-/-+vehicle;(3) STZ-ApoE-/-+vehicle;(4) STZ-ApoE-/-+Bex (10 mg· kg-1· d-1);(5) STZ-ApoE-/-+Cal (10 μg/kg, twice a week);(6) STZ-ApoE-/-+Bex (10 mg· kg-1· d-1) +Cal (10 μg/kg, twice a week).Intraperitoneal injection of STZ was performed to establish the diabetic animal model .Western blotting and immunohistochemical method was used to detect NF-κB level in the thorac-ic aorta.Plaque area in the thoracic aorta was measured using HE staining .RESULTS:Compared with the C57 mice, the fasting blood glucose in the ApoE-/-mice was not remarkably changed .The levels of total cholesterol ( TC) and low-densi-ty lipoprotein ( LDL) were greatly increased .The fasting blood glucose and lipid levels in STZ-ApoE-/-group were much higher than those in ApoE-/-group.Compared with STZ-ApoE-/-group, the fasting blood glucose and lipid levels in Bex group and Cal group were not significantly changed .Compared with the C57 mice, the protein expression of NF-κB in the ApoE-/-mice and the STZ-ApoE-/-mice was remarkably increased .Compared with STZ-ApoE-/-group, the levels of NF-κB in Bex group, Cal group and combination group were greatly decreased .Compared with STZ-ApoE-/-group, the thoracic artery plaque areas in Bex group and Cal group were inhibited (both P<0.05).Compared with Bex group, the plaque area of the thoracic artery in combination group was significantly decreased (P<0.05).CONCLUSION:Bexaro-tene or calcitriol decreases the development of atherosclerosis in streptozotocin -induced diabetic ApoE-/-mice.Bexarotene combined with calcitriol affords greater protection than monotherapy .The mechanism may be involved in down-regulating the expression of NF-κB.

AIM:To explore the effects of atorvastatin (Atorv) on atherosclerosis in streptozotocin (STZ)-in-duced diabetic apolipoprotein E knockout ( ApoE-/-) mice with fat-rich diet and the possible mechanism .METHODS:C57 mice served as control.ApoE-/-mice (n=34) fed with high-fat diet were randomly divided into ApoE-/-group, STZ-ApoE-/-group and STZ-ApoE-/-+Atorv group.Intraperitoneal injection of streptozotocin was performed to create di-abetic animal model .Blood glucose was determined by glucose oxidase method .Blood lipid levels were detected by enzymic method or selective homogeneous method .The plaque area in the thoracic aorta was measured by HE staining .The protein level of nicotinamide-adenine dinucleotide phosphate ( NADPH) oxidase subunit gp91phox in the thoracic aorta was deter-mined by Western blotting .The levels of reactive oxygen species ( ROS) in blood and thoracic aorta homogenates were de-tected by Fenton reaction and Griess reagent .Human umbilical vein endothelial cells ( HUVECs ) were isolated from healthy umbilical cords by collagenase I and cultured .ROS production was detected by flow cytometry .NADPH oxidase ac-tivity was measured using lucigenin assay .Effects of retinoid X receptor α( RXRα) on inhibition of oxidative stress by ator-vastatin were evaluated by RNA interference and plasmid transfection .RESULTS: (1) Compared with C57 group, the plaque areas of the thoracic aorta in ApoE-/-group were increased .No difference of the fasting glucose between the 2 groups was observed.The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in ApoE-/-group than those in C57 group.(2) Compared with ApoE-/-group, the plaque areas of the thoracic aorta in STZ-ApoE-/-group were further enlarged [(314.13 ±35.72) μm2 vs (215.88 ±34.19) μm2, P<0.05].The levels of blood glucose, TG, TC and LDL-C, thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in STZ-ApoE-/-group than those in ApoE-/-group (P<0.05).(3) Compared with STZ-ApoE-/-group, the plaque areas of the thoracic aorta in STZ-ApoE-/-+Atorv group were reduced [(217.47 ±24.56) μm2 vs (314.13 ±35.72) μm2, P<0.05].The levels of blood glucose , LDL-C, TC, HDL-C and TG showed no significant difference between the 2 groups.Thoracic aorta gp91phox protein level and ROS production in blood and thoracic aorta homogenates were lower in STZ -ApoE-/-+Atorv group than those in STZ-ApoE-/-group (P<0.05).(4) High glucose-induced increases in NADPH oxidase activity and gp91phox expression were significantly inhibited by atorvastatin (10-6 mol/L) in HUVECs.The inhibitory effects of atorvasta-tin on high glucose-induced ROS production and NADPH oxidase activation were largely impaired when the cells were trans -fected with RXRαsiRNA.However , the effect of atorvastatin was significantly strengthened when RXRαwas over-expressed in the HUVECs transfected with RXRαplasmid.CONCLUSION: Atorvastatin inhibits atherogenesis by depressing high glucose-induced oxidative stress in diabetic ApoE-/-mice with fat-rich diet.The anti-oxidative stress effect of atorvastatin is mediated by RXRα.

Objective To explore the effect of retinoid X receptor (RXR) agonist bexarotene on atherosclerosis and the potential mechanism in streptozotocin (STZ) induced diabetic apolipoprotein E knockout (apoE-/-) mice.Methods Eight C57BL/6 mice served as control,46 apoE-/-mice were randomized into 4 groups:apoE-/-group (n =10),STZ + apoE-/-group (n =12),STZ + apoE-/-+ Bex 10 (10 mg · kg-1 · d-1)group (n=12),STZ+ apoE-/-+Bex 30 (30 mg · kg-1 · d-1)group (n=12).Diabetic apoE-/-animal model was established by intraperitoneal injection of STZ.Blood glucose was determined by glucose oxidase method.Patch area in thoracic aorta was measured by HE staining.Western blotting was used to determine the RXR and gp91 phox protein level in thoracic aorta.Reactive oxygen species (ROS) level in blood and thoracic aorta homogenates was detected by Fenton and Griess method.Results (1) Patch areas of thoracic aorta were larger in apoE-/-group than in C57BL/6 group[(38.40 ± 8.95) μm2 vs.(0.10 ±0.01) μm2,P ＜0.01],further increased in STZ + apoE-/-group [(94.06 ± 8.04) μm2,P ＜0.05 vs.apoE-/-group] and significantly reduced in STZ + apoE-/-+ Bex 10 group [(78.72 ± 4.62)μm2,P ＜0.05 vs.STZ + apoE-/-group] and further educed in STZ + apoE-/-+ Bex 30 group[(46.13 ±7.56) μm2,P ＜ 0.05 vs.STZ + apoE-/-+ Bex 10 group].(2) Blood glucose level,TG,TC,LDL-C,thoracic aorta gp91 phox protein level and ROS level in blood and thoracic aorta homogenates were significantly higher in STZ + apoE-/-group than in apoE-/-group (all P ＜ 0.05).Blood glucose level and TG,TC,LDL-C levels were similar between STZ + apoE-/-+ Bex10 and STZ + apoE-/-group.Thoracic aorta gp91 phox protein level and ROS level in blood and thoracic aorta homogenates were lower in STZ + apoE-/-+ Bex 10 group than in STZ + apoE-/-group (P ＜ 0.05).Blood glucose level,TG,TC,LDL-C levels,gp91 phox expression in thoracic aorta,ROS level in blood and thoracic in STZ + apoE-/-+ Bex 30 group were lower than in STZ + apoE-/-group (all P ＜ 0.05).Conclusion Bexarotene treatment could attenuate arteriosclerosis progression in STZ induced diabetic apoE-/-mice,the underlying mechanism might be related to suppressing oxidative stress and decreasing blood glucose level and improving lipids metabolism.

Objective To investigate the effect of pyrrolidine dithiocarbamate(PDTC)on Pseudomonas aeruginosa induced pneumoneia of rats.Metheds A PA induced pneumonia model was established in Sprague-Dawley(SD)rats.Sixty minutes before PA exposure,PDTC and normal saline were injected into abdominal cavity separately.At 3,9,and 24 h after PA exposure,the rats were sacrificed,and the wet/dry ratio(W/D)of lung tissue was measured and histopathologic changes were observed.The number of in-filtrated polymorphonuclear(PMN)leukocytes was calculated.Immunohistochemical staining with activated NF-?B antibody was performed to detect the expression of NF-?B in lung tissues.The changes of IL-8 levels in bronchoalveolar lavage were identified by ELISA.Results Histology findings demonstrated that PA exposure induced obvious changes in lung structure,and edema and pronounced inflammatory cells infiltration were observed.Both symptoms and lesions of lung were lesser in the rats of PDTC group than those of the PA group.Compared with PDTC group,the activation of NF-?B and the expression of IL-8 were significantly up-regulated after PA challenge 3~24 h(P