1.Impact of inflammatory response on neurogenesis after cerebral ischemia
International Journal of Cerebrovascular Diseases 2011;19(10):797-800
Nerve stem cells have the potential of self-repair and multi-differentiation.They participate in brain tissue repair after ischemic stroke.The inflammatory factors have multiple functions and can change the microenvironment of brain tissue.This article reviews the recent progress in research on the inflammation after ischemic stroke and neurogenesis,mainly focusing on the impact of inflammatory factors on stem cell survival and neurogenesis.
2.Caveolin-1 and inflammation after cerebral ischemia
Sufen HUANG ; Jian YI ; Baiyan LIU
International Journal of Cerebrovascular Diseases 2016;24(3):259-262
As a cavola marker protein, caveolin-1 participates in many pathophysiological processes through its scaffolding domain oligomering many celular signal transduction molecules, and also regulates inflammation after cerebral ischemia through different pathways. This article reviews advances in caveolin-1 and inflammation after ischemic stroke in recent years, mainly focusing on its mechanism in regulating inflammation.
3.Influence of Ultra-micro-LiuweiDihuang Decoction on GH and SS of Intrauterine Growth Retardation Mice
Xiaoping PAN ; Baiyan LIU ; Guangxian CAI
Journal of Medical Research 2006;0(05):-
Objective To observe the effect of Ultra-micro-LiuweiDihuang Decoction on the embryo brain development and the influence on brain growth hormone(GH),Somatostatin(SS)in prengnant mice of passive smoking,then to demonstrate the relationship between brain and kidney in TCM.Methods Animals were divided into 5 groups at random:normal group(A),model group(B),traditional Liuwei Dihuang Decoction group(C),1/3 dosage of Ultra-micro-LiuweiDihuang Decoction group(D)and all dosage of Ultra-micro-LiuweiDihuang Decoction group(E).The IUGR model was established by passive smoking.On the 19th day of gestation,all mice were anatomized to scale the body and brain weight of lively embryos,then a part of brain tissue was homogenated.The levels of GH and SS in brain tissues were measured by ELISA.Results Passive smoking could decrease the body and brain weight,influence the level of GH and SS in brain tissues.Compared with A and B groups,the C,D and E groups could increase body and brain weight,regulate the level of GH and SS,improve the brain development of IUGR mice obviously(P
4.Effects of Buyang Huanwu decoction on the expression of Cyclin D1 and Cdk2 in rats with post-stroke depression
Juhua HOU ; Yuanyue LIU ; Can LI ; Shuhua DENG ; Baiyan LIU
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(8):680-683
Objective To observe the effects of Buyang Huanwu decoction(BYHW) on the expression of Cyclin D1 and Cdk2 in rats with post-stroke depression(PSD).Methods The rats model of focal cerebral ischemia was established by means of middle cerebral artery occlusion(MCAO).Then three weeks of salute-living and chronic unpredictable mild stress(CUMS) was given to the animal after cerebral stroke to induce the post-stoke depression animal model.The rats were divided into 5 groups:the sham operated group,the midge cerebral artery occlusion(MCAO) group,the PSD group,the fluoxetine group and the BYHWD group.The rats were subjected to left MCAO rebuilding in consistent focal cerebral ischemia,followed by an 21-day exposure to chronic mild stress (CMS)and single housing to induce PSD animal model.All rats were killed in 7,14 and 21 day after operation.The expressions of Cyclin D1 and Cdk2 were measured by immunohistochemical staining.Results Pathological changes such as hippocampal nerve cell regression,degeneration and necrosis were observed in the model rats compare with the sham operated group.The expression of Cyclin D1 in normal hippocampus in 7,14 or 21 day after operation was (1.16±0.34)%,(1.62±0.29)% and (1.60±0.24)% respectively,and Cdk2 was (1.52±0.26)%,(1.85±0.23)% and (1.97±0.10)%.After PSD the expression of Cyclin D1 was (49.69±5.68)%,(58.17± 2.89) % and (50.87 ± 2.48) % respectively,and Cdk2 was (50.63 ± 2.93) %,(70.34± 1.47) % and (61.35 ± 3.04) %.Compared with model group,Fluoxetine and BYHW significantly reduced the numbers of Cyclin D1 and Cdk2 positive cells,the expression of Cyclin D1 was (16.62±4.27)%,(29.66±5.24)% and (26.71±1.32)% at fluoxetine group,and Cdk2 was (18.05±2.26) %,(33.84±3.12) % and (24.51±2.66) %.The expression of Cvclin D1 was (14.62±2.06)%,(26.89±3.41)% and (23.68±2.01)% at BYHWD group,and Cdk2 was (16.60± 2.42) %,(20.09±3.45) % and (22.19± 1.70) %.Conclusion The abnormal expression of Cyclin D1 and Cdk2 at the PSD rats indicate that they may be involved in the mechanism of neuronal death.Buyang Huanwu decoction may reduce the neuronal apoptosis through down-regulating the expression of Cyclin D1 and Cdk2.
5.Effect of Buyang Huanwu Decoction on vascular endothelial growth factor and its receptor Flk1 in rats after focal cerebral ischemia
Baiyan LIU ; Guangxian CAI ; Wei LIU ; Xuemei CHEN
Chinese Traditional and Herbal Drugs 1994;0(03):-
Objective To explore the effect of Buyang Huanwu Decoction (BYHWD) on vascular endothelial growth factor (VEGF) and its receptor, Fetal liver kinase 1 (Flk1) in rats after focal cerebral ischemia. MethodsRats were randomly divided into following groups including Sham group, model group, Nimodipine group, and BYHWD group. The model of focal cerebral ischemia in rats was repro-duced by middle cerebral artery occlusion. Rats were ig administered and killed after operated, then the expression of VEGF and Flk1 and protein level of VEGF in brain tissue of every groups were measured by immunohistochemisty and enzyme linked immunosorbent assay, and the nervous function deficit scores were evaluated. ResultsThere were a few VEGF and Flk1 positive cells in normal brain tissue of rats. After focal cerebral ischemia the VEGF and Flk1 positive cells were increased. Compared to model group, Nimodipine and BYHWD significantly improved the neurological behavior performance, increased the numbers of VEGF and Flk1 positive cells, and enhanced the VEGF protein level (P
6.Effect of Bu Yang Huan Wu decoction(补阳还五汤) on basic fibroblast growth factor in rats with cerebral ischemia
Fang LIU ; Xuesong BAI ; Baiyan LIU ; Guangxian CAI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(01):-
Objective To investigate the effect of Bu Yang Huan Wu decoction(BYHWD,补阳还五汤) on the expression of basic fibroblast growth factor(bFGF) and its mechanism of anti-cerebral ischemia.Methods Eighty-five Sprague-Dawley rats(body weight 280-300 g) were randomly divided into five groups: normal control group(n=5),sham-operated group,model group(n=20),traditional BYHWD group and ultra-fine powder BYHWD group.Then the latter four groups were further divided into four subgroups of 1,3,5 and 7 days(each n=5).The cerebral ischemic rat model was made by occluding the middle cerebral artery(MCA) with a thread.The traditional decoction and ultra-fine powder decoction groups were ingested with the liquid medicine of BYHWD after 2 hours of the operation(2 ml per rat per day,according to the surface area,it was equivalent to three times the dosage of 70 kg adult(human)).All groups were executed at the 1,3,5 and 7 days after the operation.At the same time there were 5 rats as the normal control group.The brain tissue was taken for the preparation of specimens.The distribution and expression difference of bFGF in the ischemic brain tissues of rats in different groups had been investigated with immuohistochemistry technique.The contents of bFGF in the brain tissue were detected with enzyme linked immunosorbent assay(ELISA).Results The positive expression of bFGF at peri-infarct area after ischemia increased obviously,but at central-infarct area there was less such positive expression.There was no expression at contra-lateral side to the ischemic area.The expression of bFGF immune positive cell increased at 1 day,peaked at 3 days,and returned to baseline at 7 days.The positive cells of bFGF in tradition decoction and ultra-fine powder decoction groups were obviously more than those in the model group at 3,5 and 7 days(all P
7.Effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide in chronical stressed mice
Baiyan LIU ; Jian YI ; Xuemei CHEN ; Xiaoyuan LIN ; Guangxian CAI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(4):292-294
Objective To explore effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide(VIP) in the hypothalamus, spinal cord and duodenum of chronical stressed mice. Methods Mice were randomly divided into normal, stress and Simotang group( n= 10 in each group), and given a variety of unpredictable chronic mild stress. After 21 days gastric emptying and intestinal propulsion function were measured,the expression of VIP was detected by immunohistochemistry and RT-PCR. Results Compared with mice in normal group( (49.81 ± 8.56)%; (51.02 ± 5.11 )% ), chronic stress increased gastric residual rate( (61.53 ±8.71 ) %; P < 0.05 ) and reduced small intestine propulsion rate ( ( 31.79 ± 2.38 ) %; P < 0. 05 ). There were differences in expressions of VIP positive cells and mRNA in duodenum( (8.8 ± 1.1 )/mm2 and(0. 58 ±0.03) ),hypothalamus ( ( 12.9 ± 1.5 )/mm2 and (0.81 ± 0. 07 ) ) and spinal cord ( ( 12.1 ± 1. 2)/mm2 and (0.76 ± 0.02) )in chronic stress group compared with normal group( (6.5 ± 0. 9)/mm2 and (0.43 ± 0. 04);( 10.8 ± 1.3 )/mm2and (0.57 ± 0.03 ); (9.3 ± 1.5 )/mm2 and (0.53 ± 0. 02 ) respectively). There was not difference in gastric residual rate (52.93 ± 9.15 )%, small intestine rate(48.98 ± 4.38 )% and expressions of VIP positive cells and mRNA in duodenum ( (6.7 ± 0.9)/mm2 and (0.48 ± 0. 05 ) ), hypothalamus ( ( 10. 6 ± 1.4 )/mm2 and ( 0. 61 ± 0. 05 ) )and spinal cord ( (9. 1 ± 1.3)/mm2 and(0.55 ± 0.05 ) ) in Simotang group compared with those in normal group (P > 0.05 ), but there were decreased compared with those in chronic stress group (P < 0.05 ). Conclusion Simotang can regulate expressions of VIP in duodenum, hypothalamusand spinal cord in chronically stressed mice.
8.Genotype identification and breeding method of caveolin-1 gene knockout mice
Shengqiang ZHOU ; Dong LUO ; Sufeng HUANG ; Jian YI ; Baiyan LIU
Acta Laboratorium Animalis Scientia Sinica 2016;24(3):228-232
Objective To investigate the identification and optimal breeding method of caveolin-1 knockout mice, and provide an ideal animal model for further study of the role of caveolin-1 in cerebral ischemic injury and repair. Meth?ods The introduced caveolin-1 gene knockout mice were reared in the SPF laboratory and genomic DNA was extracted from mouse tail tissue by the method of boiling lysis. According to the primer sequences provided by the Jackson Laboratory of America for polymerase chain reaction ( PCR) to detect the genotypes, with the four different ways of mating:caveolin-1 +/ -heterozygote intercrossing, heterozygous and homozygous caveolin-1 -/ -hybrid ( orthogonal and pay) as well as homo-zygous intercrossing. The pregnancy rate, shape characteristics of the filial generation mice and homozygous rate of the pa-rental mice were observed. Results Agarose gel electrophoresis results indicated that the size of molecular weight of the PCR products was about 200 bp and 661 bp, which were consistent with the expected target gene fragment, and identified caveolin-1 gene knockout mice of different genotypes successfully. The results of different mating patterns are basically in a-greement with Mendel rule, and the female and male aveolin-1 -/ -homozygous mice had a certain ability to reproduce, three different genotypes of mice had no significant differences between the shape features. Conclusions PCR can fast and reliably identify the genotypes of caveolin-1 knockout mice using genomic DNA through the method of boiling lysis. Combi- ning the breeding methods of intercrossing of caveolin-1 heterozygous mice and intercrossing of caveolin-1 homozygous mice may be a good way to obtain enough homozygous mice and homologous wild type mice in a short period.
9.Culture and biological characteristics of neural stem cells from caveolin-1 knockout embryonic mice
Baiyan LIU ; Yue YU ; Jian YI ; Xuemei CHEN ; Guangxian CAI
Chinese Journal of Tissue Engineering Research 2014;(23):3739-3744
BACKGROUND:Caveolin-1 is expressed in mammalian brain and involved in the normal development of the brain, which can affect the proliferation of neural stem cells in the brain. OBJECTIVE:To acquire neural stem cells from caveolin-1 knockout embryonic mice in vitro and study their biological characteristics. METHODS:The whole brain was separated from C57BL/6 mice and caveolin-1 knockout C57BL/6 mice respectively at encyesis 14-16 days. Single cellsuspension was obtained by enzyme digestion, and cultured in the conditioned medium of neural stem cells. Fol owing 7 days of primary culture, the cells were induced in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum for 7 days. RESULTS AND CONCLUSION:The major cells of the cellsuspensions from the fetal mouse brain were dead at 1 day after culture, and some single cells floated in the medium and their transmittance were better, and then they gradual y formed multicellular bal s after 3 days. A smal amount of cells were adhered at the bottom of culture plate after passage, and a great amount of cellbal s appeared after 7 days. The proliferation rate of neural stem cells from caveolin-1 knockout mice was higher than that from normal mice. The cellbal s were nestin-positive and their differentiated cells was positive for neurofilament 200, glial fibril ary acidic protein or O4, respectively. Al of the cells from normal mouse brain were positive for caveolin-1, but the cells from caveolin-1 knockout mice were negative for caveolin-1 by immunocytochemistry. Moreover, the speed of cellbal formation and the number of cellbal s in neural stem cells from caveolin-1 knockout mice were better than those from normal mice. Caveolin-1 negative neural stem cells were cultured successful y from caveolin-1 knockout mouse brain, and the results show that caveolin-1 can promote the proliferation of neural stem cells and inhibit their differentiation in vitro.
10.Effects of caveolin-1 on expressions of interleukin-1βand interleukin-6 in the ischemic cortex after permanent middle cerebral artery occlusion in mice
Sufen HUANG ; Shengqiang ZHOU ; Dong LUO ; Jian YI ; Baiyan LIU
International Journal of Cerebrovascular Diseases 2016;24(11):1022-1027
ObjectiveToinvestigatetheeffectsofcaveolin1(Cav1)onexpressionsofproinflammatory cytokine interleukin (IL)-1βand IL-6 in the ischemic cortex after permanent middle cerebral artery occlusion in mice. Methods The Cav-1 knockout mice (n=40) and wild-type mice (n=40) were randomly divided into ischemia groups and sham operation groups (n=20 in each group). They w ere also redivided into ischemia or sham operation at 3, 7, 10 and 14 d time points ( n=5 in each time point). A permanent middle cerebral artery occlusion model w as induced by the suture method. Immunohistochemical method w as used to detect the expressions of IL-1βand IL-6 in the ischemic cortex. Results The expression levels of IL-1βand IL-6 in the ischemic cortex at each time point in the ischemia group in Cav1 knockout mice w ere significantly higher than those in the ischemia group in the w ild-type mice ( al P< 0.05 ). Conclusions The upregulations of proinflammatory cytokines IL-1βand IL-6 in the ischemic cortex in Cav1 knockout mice suggests that Cav1 plays an important role in aleviating inflammation after cerebral ischemia.