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BACKGROUND: A variety of stem cells have been found to be effective in the treatment of Alzheimer's disease in rats. However, few reports have been reported on the treatment of Alzheimer's disease rats with brain-derived neurotrophic factor (BDNF)-modified human amniotic membrane-derived mesenchymal stem cells. OBJECTIVE: To investigate the effects of BDNF-modified human amniotic membrane-derived mesenchymal stem cell transplantation on the learning and memory abilities of Alzheimer's disease rats. METHODS: Forty-eight Sprague-Dawley rats were divided into control group (no treatment), model group (Alzheimer's disease model), stem cell transplantation group (human amniotic membrane-derived mesenchymal stem cell transplantation+Alzheimer's disease model) and BDNF-modified stem cell transplantation group (BDNF-modified human amniotic membrane-derived mesenchymal stem cell transplantation+Alzheimer's disease model), 12 rats in each group. Learning and memory of model rats were determined in a trisection radiation maze and immunohistochemical staining was used to determine the number of p75 positive neurons at 2 weeks after cell transplantation. RESULTS AND CONCLUSION: The number of p75 positive neurons in the bevel zone and medial septal nucleus was ranked as follows: the model group < the stem cell transplantation group < the BDNF-modified stem cell transplantation group < the control group, and there were significant differences among groups (P < 0.05). The learning and memory abilities of the rats were ranked as follows: the model group < the stem cell transplantation group < the BDNF-modified stem cell transplantation group < the control group, and there were significant differences among groups (P < 0.05). In the BDNF-modified stem cell transplantation group, the number of learnings was negatively correlated with the number of p75 NGFR-positive neurons (P < 0.05), while the memory capacity was positively correlated with the number of p75 NGFR-positive neurons (P < 0.05). These findings reveal that human amniotic membrane-derived mesenchymal stem cell transplantation can improve learning and memory abilities of Alzheimer's disease rats, and BDNF-modified human amniotic membrane-derived mesenchymal stem cells can further improve this therapeutic effect.
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Objective·To investigate the effects of insulin-like peptide 6 (Insl6) on renal fibrosis and calcification in unilateral ureteral obstruction (UUO) mice. Methods·Twenty-four SPF male mice with genotypic background of C57BL/6 were divided into Sham (n=8), UUO+saline (n=8) and UUO+Insl6 (n=8) groups randomly. Mice were sacrificed 10 days after operation and renal tissues of surgical side were obtained. Sirus red staining, Masson staining and alizarin red S staining were used to verify the level of collagen and calcium deposition. TGF-β1 expression was determined by Western blotting. Realtime-PCR was used for determining TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression. Results·Compared with sham group, fibrotic area especially collagen Ⅰ , calcium deposition, TGF-β1 protein, and TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression in UUO+saline group significantly increased (all P<0.05). As compared with UUO+saline group, fibrotic area especially collagen Ⅰ, calcium deposition, TGF-β1 protein, and TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression in UUO+Insl6 group significantly decreased (all P<0.05). Conclusion·Insl6 inhibits UUO-induced renal fibrosis and calcification, which may be related to regulation of TGF-β1, collagen Ⅰ , BMP2 and collagen Ⅱ expression levels.
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Objective To investigate the regulative effect of exogenous Angiotcnsin-(1-7) [Ang(1-7)] on nuclear factor-κB (NF-κB) and its downstream proinflammatory cytokines in rats after cerebral ischemic-reperfusion injuries.Methods Forty-two male SD rats were randomly divided into sham-operated group,control group and Ang-(1-7) treatment group (n=14); cerebral ischemia in control group and Ang-(1-7) treatment group was induced by transient middle cerebral artery occlusion (MCAO).The rats in the sham-operated group and control group were infused with artificial cerebrospinal fluid (aCSF,0.5 μL/h) while rats in the Ang-(1-7) treatment group were with Ang-(1-7)(100 pmol,0.5 μL/h)into the lateral ventricle by implanted osmotic minipumps following reperfusion.At 24 h after reperfusion,all rats were sacrificed to detect the expression of NF-κB p65 subunits in cell nuclei of the ischemic cortex by Western blotting.The spatial distribution of NF-κB p65 subunits in ischemic cerebral tissues was detected by immuno-histochemical assay.The concentrations of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum were detected by ELISA.Results A significant reduction of NF-κB p65 expression in cell nuclei by 46％ in the Ang-(1-7) treatment group was noted as compared with that in control group (P＜0.05).The nuclear translocation of NF-κB p65 in the ischemic cortical cells in the Ang-(1-7) treatment group was significantly reduced,and the rate of NF-κB p65 positive cells was decreased by 29％ as compared with that in control group (P＜0.05).Besides,the serum levels of TNF-α (71.603±18.539 pg/mL) and IL-1 β (44.648±10.387 pg/mL) in the Ang-(1-7) treatment group were obviously decreased as compared with those in the control group (104.763±24.412 pg/mL,64.787±14.441 pg/mL,P＜0.05).Conclusion Exogenous administration of Ang-(1-7) could attenuate inflammatory reaction following cerebral ischemia,perhaps by interacting with Mas receptor or through the antagonism against the pro-inflammatory effect of Ang Ⅱ.
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Objective To investigate the relationship between NINJ2 gene polymorphism and stroke, and the differences of serum levels of tumor necrosis factor-αt (TNF-α), NGF, interleukin-6 (IL-6)and P-Selectin in healthy controls and patients under recovery stage. Methods Fifty-two patients with large-artery atherosclerosis (LAA) infarction, 85 patients with small-artery occlusion lacunar (SAO)infarction, 50 patients with intracerebral hemorrhage (ICH) and 66 healthy controls were included in this study. Genotypes of the 2 single nucleotide polymorphism (SNP) sites (rs12425791 and rs11833579) in NINJ2 gene were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RLFP) method. The differences of genotypes and alleles frequencies of the 2 SNP sites between each 2 different groups were analyzed and compared. Multinomial logistic regression was used to calculate the odds ratio (OR) of genotypes in each patient group, and 95% confidential interval (95% CI)was given. The serum levels ofTNF-α, NGF, IL-6 and P-Selectin were tested by ELISA method, and compared between groups and within group classified by genotypes. Results In regard to rs12425791 site, the frequencies of AG and AA+AG genotypes in LAA and SAO groups were significantly higher than those in control group (P＜0.05), while this difference was not found between the ICH group and control group (P＞0.05); the frequency of A allele in the SAO group was significantly higher than that in the control group (P＜0.05), while this difference was not found between the control group and both the LAA and ICH groups (P＞0.05). In regard to rs11833579 site, no significant differences in the genotypes and alleles were noted between all the patient groups and control group (P＞0.05). After adjusting the influence of other risk factors, the multinomial logistic regression analysis showed that the onset of stroke was still significantly associated with the AG genotype at rs12425791 site in the LAA group (OR=4.298,95%CI=1.430-12.922) and AG, AG+AA genotypes at rs12425791 site in the SAO group (OR=3.923 and 2.937, 95%CI= 1.417- 10.860 and 1.119-7.710). Neither genotypes in rs 11833579 site were significantly associated with the onset of stroke. No significant differences of serum levels of TNF-α, NGF, IL-6 and P-Selectin were noted between each patient group under recovery stage and control group (P＞0.05); in regard to rs12425791 site, the serum level of TNF in LAA group with different genotypes was significantly different (P＜0.05) and the serum level of P-Selectin in ICH group with different genotypes was significantly different (P＜0.05); in regard to rs11833579 site, the serum level of NGF in LAA group with different genotypes was statistically different (P＜0.05). Conclusion This SNP site (rs12425791)is significantly associated with ischemia stroke and the A allele increases the risk of being susceptible to this disease in Chinese Han population. That SNP site (rs1 1833579) is not significantly associated with stroke. No significant differences of TNF-α, NGF, IL-6, P-Selectin serum levels are noted between patients under recovery stage and controls.
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Objective To clarify the relation between the C3435T polymorphism of multidrug resistance 1 (MDR1) gene and human refractory epilepsy (RE) in ethnic Han Chinese. Methods We collected 170 patients with epilepsy, whose diagnoses were correct and treatments were reasonable. RE was defined as having uncontrolled seizures that occurred with an average frequency of at least once a month for a period of at least 2 years; during the 2-years period, at least 2 different antiepileptic drugs (AEDs) were used daily, either singly or in combination. According to the definition, 91 patients were classified into RE group and the other 79 patients into non-RE group. A 5-mL venous blood sample was taken from the patients for DNA extraction and genotyping. Genotype of C3435T polymorphism in MDR1 gene was determined by polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP). Results The distribution of CC, CT, TT genotypes was 48.4%, 40.7%,11.0% in RE group, and 40.5%, 38.0%, 21.5% in non-RE group, respectively; no significant differences of C3435T genotype were noted between the 2 groups (x2=3.615, P=0.164). The C and T allele frequencies were 68.7%, 31.3% in RE group, and 59.5%, 40.5% in non-RE group, respectively; no significant differences were found between 2 groups (x2=3.112, P=0.080). Patients were divided into primary epilepsy group and cryptogenic or symptomatic epilepsy group according to the etiology;analyses of the genotype and allele of C3435T in the sub-groups (RE and non-RE subgroups) of this 2 groups were similarly unremarkable. Conclusion No association between the C3435T polymorphism in MDR1 gene and RE in ethnic Han Chinese is noted.
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Objective To investigate the protective effects ofAngiotensin-(1-7) [Ang-(1-7)] against the focal cerebral ischemic-reperfusion injury in rats. Methods Spragne-Dawley rats were randomly divided into sham operated group, Ang-(1-7) treated group and aCSF treated group. The latter 2 groups were subjected to middle cerebral artery occlusion (MCAO). The 3 groups were administrated artificial cerebrospinal fluid (aCSF, 0.5 μL/h), Ang-(1-7) (100 pmol, 0.5 μL/h) and aCSF 0.5 μL/h,respectively, by implanted Alzet osmotic minipumps into lateral cerebral ventricle at reperfusion 24 h and 48 h. In all experimental rats, their neurological function scores, the brain edema at reperfusion 48 h and the cerebral infarct size at reperfusion 24 h were evaluated. And the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the ischemic cerebral tissue at reperfusion 24 h and 48 h were also determined. The number of apoptotic neurons within the tissue around the infarct at reperfusion 48 h was detected by the way of staining with terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick-end labeling (TUNEL). Results In the treatment of MCAO rats, Ang-(1-7) significantly ameliorated their neurological function score (P＜0.05), reduced the infarct size (P＜0.05), decreased the tissue MDA content (P＜0.05), increased the tissue SOD activity (P＜0.05). It also reduced markedly the number of apoptotic neurons around the infarct (P＜0.01), but had no effect on the water content in the brain. Conclusions Ang- (1-7) has a neuroprotective effect against cerebral ischemic-reperfusion injury, perhaps by its anti-oxidation stress and inhibition of neuronal apoptosis.
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<p><b>OBJECTIVE</b>To analyze the role of resistin in insulin resistance (IR) through investigating the variation of plasma resistin levels and single-nucleotide polymorphisms (SNPs) in resistin gene 5' flanking region in stroke patients.</p><p><b>METHODS</b>In 103 atherothrombotic cerebral infarction (ACI) patients, 85 lacunar infarction (LI) patients, 70 intracerebral hemorrhage (ICH) patients, and 86 healthy controls, plasma resistin and insulin levels were measured by ELISA, SNPs in resistin gene 5' flanking region were detected by PCR and direct DNA sequencing. The subjects' body height and weight, the body mass index, quantitative insulin sensitivity check index (QUICKI), blood pressure, and the concentration of fasting plasma glucose, triglyceride, total cholesterol, creatinine, low-density lipoprotein, and high-density lipoprotein were also determined.</p><p><b>RESULTS</b>QUICKI was significantly lower in the ACI and ICH patients (0.316 +/- 0.037 and 0.309 +/- 0.032, respectively) than that in the controls (0.342 +/- 0.043, P < 0.001), while plasma resistin level was significantly higher in the ACI and ICH patients (6.36 +/- 3.79 and 7.15 +/- 4.27 ng/mL, respectively) than that in the controls (5.28 +/- 2.56 ng/mL, P < 0.05), but such difference was not observed in the LI patients compared with controls. There was a statistically negative correlation between plasma resistin level with QUICKI (r = -0.228, P < 0.001). The distributions of allele and genotype frequencies of resistin gene - 420C > G and - 537A > C SNPs were not significantly different among the different groups, and those SNPs were not correlated with other clinical and biochemical parameters.</p><p><b>CONCLUSIONS</b>Plasma resistin is associated with stroke by participating in the development of IR. The SNPs in resistin gene 5' flanking region has no impact on the plasma resistin level.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Blood Glucose , Metabolism , Cerebral Infarction , Blood , Genetics , Creatinine , Blood , Insulin , Blood , Insulin Resistance , Physiology , Intracranial Arteriosclerosis , Blood , Genetics , Lipoproteins , Blood , Polymorphism, Single Nucleotide , Resistin , Blood , Genetics , Stroke , Blood , Genetics , Triglycerides , BloodABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship of plasma homocysteine (Hcy) levels and the gene polymorphisms of N5, N10-methylenetetrahydrofolate reductase (MTHFR), cystathionine beta-synthase (CBS) with Alzheimer's disease (AD).</p><p><b>METHODS</b>Plasma Hcy levels were measured by means of high voltage capillary electrophoresis with ultra-violet detection, the polymorphisms of C677T in exon 4 of MTHFR gene and 844ins68 in exon 8 of CBS gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 105 AD patients and 102 non-AD controls. All controls were excluded from cardiocerebrovascular disorders and other diseases.</p><p><b>RESULTS</b>The plasma Hcy level in AD patients (16.04 +/- 3.84 micromol/L) was significantly higher than that in the controls (11.94 +/- 3.87 micromol/L, P < 0.001). There were no significant differences of the genotype and allele frequencies of MTHFR C677T mutation and CBS 844ins68 mutation between the patients and controls. However, the T allele of MTHFR gene was found to relate with the plasma Hcy level increase in all subjects.</p><p><b>CONCLUSION</b>The elevated plasma Hcy level in AD patients is probably involved in the pathogenesis of AD, which may be due to the environmental factor rather than genetic factors of the mutations of MTHFR and CBS.</p>