Betaine Inhibits NLRP3 Inflammasome Hyperactivation and Regulates Microglial M1/M2 Phenotypic Differentiation, Thereby Attenuating Lipopolysaccharide-Induced Depression-Like Behavior.

Depression is one of the most important mental illnesses and is closely related to inflammation. Betaine is a natural product with an anti-inflammatory and antioxidant activities. However, the mechanism by which betaine ameliorates depression-like behaviors induced by lipopolysaccharide (LPS) is poorly understood. The purpose of this study was to investigate the neuroprotective effect of betaine on LPS-induced depression-like behavior in mice and its mechanism of action. ICR mice were randomly divided into four groups: the control group, the LPS model group (0.83 mg/kg), the positive drug group (MIDO, 50 mg/kg), and the betaine group (5% and 1% in drinking water). The betaine group was administered for 21 days, and on the 22nd day, except for the blank group, LPS (0.83 mg/kg) was intraperitoneally injected to establish a lipopolysaccharide-induced mice depression-like model. Twenty-four hours after LPS injection, the tail suspension test (TST), open field test (OFT), and sucrose preference test (SPT) were performed to evaluate the effect of betaine on LPS-induced depressive behavior in mice. After the behavioral study, the mouse brain, hippocampus, and serum were taken for detection. The expressions of cytokines and inflammatory mediators were detected by ELISA, HE staining, immunofluorescence, immunohistochemistry, and western blotting. Western blotting was used to detect the protein expression levels of the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, and ASC, the protein expression levels of the microglial polarization markers COX-2, inducible nitric oxide synthase (iNOS), and CD206. The results showed that betaine significantly ameliorated the depression-like behavior in LPS-induced mice, significantly attenuated the production of proinflammatory cytokines and increased the release of an anti-inflammatory cytokines. Betaine decreased the expression of the NLRP3 inflammasome, decreased the expression of M1 polarization markers, tumor necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), COX-2, and iNOS and promoted the expression of M2 polarization marker CD206. Our study suggests that betaine may promote the transition of microglia from the M1 to the M2 phenotype by inhibiting NLRP3 inflammasome activation, thereby attenuating lipopolysaccharide-induced depression-like behavior.

Clinical effects and safety of electroacupuncture for the treatment of post-stroke depression: a systematic review and meta-analysis of randomised controlled trials.

OBJECTIVE: The aim of this systematic review was to assess the efficacy/effectiveness and safety of electroacupuncture (EA) in the treatment of post-stroke depression (PSD). METHODS: A comprehensive literature search in the Pubmed, Embase, CENTRAL, ISI Web of Science, CNKI and Wanfang databases was conducted, and all relevant randomised controlled trials (RCTs) were screened for eligibility by two independent reviewers. The Cochrane Collaboration's tool and Jadad score were used to assess the risk of bias of included studies, and only RCTs scoring ≥3 were included in a meta-analysis. RESULTS: 18 RCTs involving a total of 813 participants (mean age 61.6 years) in the EA groups and 723 participants (mean age 61.9 years) in the control groups were included. The included studies had an average 3 point Jadad score. PSD was diagnosed according to the Chinese Classification of Cerebrovascular Disease (CCCD) and the Chinese Classification of Mental Disease (CCMD) criteria. There was no significant difference between EA and antidepressants (fluoxetine 10-40 mg/day, citalopram 20 mg/day, sertraline 50 mg/day) in terms of the Hamilton Depression Rating Scale (HAMD) scores at week 4 after treatment (standardised mean difference (SMD) -0.11, 95% CI -0.31 to 0.10), at week 6 after treatment (SMD 0.04, 95% CI -0.43 to 0.51) or at week 8 after treatment (SMD -0.01, 95% CI -0.23 to 0.22). However, the combined incidence of adverse events in the EA groups was significantly lower than in the antidepressant groups (RR 0.21, 95% CI 0.14 to 0.33). CONCLUSION: There was no significant difference between EA and antidepressants in the severity of depression, however EA caused fewer adverse events than antidepressants. Additional larger scale RCTs with rigorous study design are required.

A SNP in 5' untranslated region of CD40 gene is associated with an increased risk of ischemic stroke in a Chinese population: a case-control study.

Cluster of differentiation 40 (CD40), the receptor for CD154, is a member of the tumor necrosis factor (TNF) receptor superfamily. Several studies have been conducted to investigate the effect of the CD40 rs1883832 polymorphism on atherosclerotic disease in different population; however, inconsistent results were obtained. In this study, we investigated the association of four polymorphisms (rs1883832, rs13040307, rs752118 and rs3765459) of CD40 gene and their effect on CD40 expression with the risk of ischemic stroke (IS) in a Chinese population. Three hundred and eighty patients with IS and 450 control subjects were included in the study. The CD40 polymorphisms were discriminated by Snapshot SNP genotyping assay. Serum soluble CD40 (sCD40) levels were detected by ELISA. We found that the rs1883832CT and rs1883832TT genotypes were associated with an increased risk of IS compared with the rs1883832CC genotype (OR = 1.42, 95% CI: 1.03-1.95, p = 0.030 and OR = 1.91, 95% CI: 1.29-2.82, P = 0.001, respectively), and the rs1883832T allele was associated with a significantly increased risk of IS compared with rs1883832C allele (OR = 1.40, 95% CI: 1.15-1.70, P = 0.001). Elevated serum sCD40 levels were observed in patients with IS compared with the control gropu (P < 0.01). Individuals carrying the rs1883832TT or rs1883832CT genotypes showed significantly higher sCD40 levels compared with the rs1883832CC genotype in the IS group [(64.8 ± 25.4 pg/mL, TT = 94); (63.9 ± 24.3 pg/mL, CT = 185) vs (53.3 ± 22.5 pg/mL, CC = 101), P < 0.01]. The TCCA haplotype was associated with an increased risk of IS compared with the control group (OR = 2.10, 95% CI: 1.23-3.58, p = 0.005). However, we did not find a significant association between the other three polymorphisms and IS risk. In conclusion, after a comprehensive comparison with other studies, we confirmed that the rs1883832T allele but not the rs1883832C allele is associated with an increased risk of IS. The rs1883832 polymorphism may exert influences on abnormal CD40 expression in IS patients among the Chinese population.

A SNP in 5′ untranslated region of CD40 gene is associated with an increased risk of ischemic stroke in a Chinese population: a case-control study

Abstract Cluster of differentiation 40 (CD40), the receptor for CD154, is a member of the tumor necrosis factor (TNF) receptor superfamily. Several studies have been conducted to investigate the effect of the CD40 rs1883832 polymorphism on atherosclerotic disease in different population; however, inconsistent results were obtained. In this study, we investigated the association of four polymorphisms (rs1883832, rs13040307, rs752118 and rs3765459) of CD40 gene and their effect on CD40 expression with the risk of ischemic stroke (IS) in a Chinese population. Three hundred and eighty patients with IS and 450 control subjects were included in the study. The CD40 polymorphisms were discriminated by Snapshot SNP genotyping assay. Serum soluble CD40 (sCD40) levels were detected by ELISA. We found that the rs1883832CT and rs1883832TT genotypes were associated with an increased risk of IS compared with the rs1883832CC genotype (OR = 1.42, 95% CI: 1.03-1.95, p = 0.030 and OR = 1.91, 95% CI: 1.29-2.82, P = 0.001, respectively), and the rs1883832T allele was associated with a significantly increased risk of IS compared with rs1883832C allele (OR = 1.40, 95% CI: 1.15-1.70, P = 0.001). Elevated serum sCD40 levels were observed in patients with IS compared with the control gropu (P < 0.01). Individuals carrying the rs1883832TT or rs1883832CT genotypes showed significantly higher sCD40 levels compared with the rs1883832CC genotype in the IS group [(64.8 ± 25.4 pg/mL, TT = 94); (63.9 ± 24.3 pg/mL, CT = 185) vs (53.3 ± 22.5 pg/mL, CC = 101), P < 0.01]. The TCCA haplotype was associated with an increased risk of IS compared with the control group (OR = 2.10, 95% CI: 1.23-3.58, p = 0.005). However, we did not find a significant association between the other three polymorphisms and IS risk. In conclusion, after a comprehensive comparison with other studies, we confirmed that the rs1883832T allele but not the rs1883832C allele is associated with an increased risk of IS. The rs1883832 polymorphism may exert influences on abnormal CD40 expression in IS patients among the Chinese population.

Apolipoprotein E polymorphisms increase the risk of post-stroke depression.

Recent reports have shown that apolipoprotein E (APOE) polymorphisms are involved in neurodegenerative disease. However, it is unclear whether APOE affects post-stroke depression. Accordingly, we hypothesized that APOE polymorphisms modify the risk of post-stroke depression. Here, we performed a hospital-based case-control study (including 76 cerebral infarction cases with post-stroke depression, 88 cerebral infarction cases without post-stroke depression, and 109 controls without any evidence of post-stroke depression or cerebral infarction) to determine possible association between APOE rs429358 and rs7412 polymorphisms and risk of post-stroke depression. Our findings show no difference among the groups with regards genotype distribution of the rs7412 polymorphism. In contrast, APOE genotypes with rs429358-C alleles increased the risk of post-stroke depression. Further, the rs429358 polymorphism was associated with significantly decreased regional cerebral blood flow values in the left temporal lobe of post-stroke depression cases. Additionally, the rs429358 polymorphism was not only associated with depression severity, but with increasing serum levels of total cholesterol. These results suggest that the APOE rs429358 polymorphism is associated with increased risk of developing post-stroke depression, and that APOE rs429358-C allele genotypes may be detrimental to recovery of nerve function after stoke. Indeed, these findings provide clinical data for future post-stroke depression gene interventions.

SNCA rs3822086 C>T Polymorphism Increases the Susceptibility to Parkinson's Disease in a Chinese Han Population.

OBJECTIVE: Protofibrils of alpha-synuclein mediate neuronal cell death and propagate Parkinson's disease (PD). In this study, we investigated the relationship between the rs3822086 C>T polymorphism located in the fourth intron of the alpha-synuclein (SNCA) gene and susceptibility to PD in a Chinese Han population. METHODS: 146 PD patients and 144 sex- and age-matched healthy individuals (control group) were selected for this study. The SNCA rs3822086 polymorphism was examined in all 300 study subjects by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: The genotype and allele frequencies of the SNCA rs3822086 polymorphism showed significant differences between the PD group and control group (TT: 25.3% vs. 18.8%, p=0.035; CT+TT: 77.4% vs. 66.0%, p=0.031; T allele: 51.4% vs. 42.4%, p=0.030; respectively). Stratified analyses based on gender indicated that male PD patients exhibited higher genotype and allele frequencies of the SNCA rs3822086 polymorphism compared to healthy male controls (TT: 26.7% vs. 13.2%, p=0.011; CC+CT: 73.3% vs. 86.8%, p=0.024; T allele: 51.2% vs. 37.9%, p=0.012; respectively). Age-stratified analyses indicated that the genotype and allele frequencies of the SNCA rs3822086 polymorphism were significantly higher in PD patients older than 60 years in comparison to healthy controls (TT: 32.2% vs. 20.5%, p=0.014; CT+TT: 77.0% vs. 60.2%, p=0.017; T allele: 54.6% vs. 40.3%, p=0.008; respectively). CONCLUSION: Our findings demonstrate that the SNCA rs3822086 C>T polymorphism correlates with increased susceptibility to PD among the Chinese Han population.

The association of L-selectin polymorphisms with L-selectin serum levels and risk of ischemic stroke.

L-selectin is a member of the selectin family of cell adhesion molecules which are important in the transient attachment of leukocytes to endothelial cells, which plays a role in inflammation processes and is one of the earliest events in the pathogenesis of atherosclerosis. No studies have examined the association of this polymorphism with ischemic stroke. Therefore, we investigated that L-selectin gene polymorphism and its soluble level are associated with ischemic stroke in Chinese population. We analyzed single nucleotide polymorphisms of L-selectin gene Pro213Ser (P213S) in 265 patients with ischemic stroke and 280 age and sex matched controls, using PCR-RFLP and DNA sequencing method, while soluble L-selectin levels were measured by ELISA. There were significant differences in the genotype and allele frequencies of L-selectin gene P213S polymorphism between the group of patients with ischemic stroke and the control group (P < 0.05). Soluble L-selectin levels were increased in patients with ischemic stroke compared with controls (P < 0.01). Moreover, The P213S polymorphism of L-selectin was significantly associated with sL-selectin levels, the serum levels of L-selectin PP genotype carriers was significantly higher than no carriers in patients with ischemic stroke (P < 0.05). The P213S polymorphism of L-selectin and its sL-selectin levels are associated with ischemic stroke in Chinese population. Our data suggests that L-selectin gene may play a role in the development of ischemic stroke.

A novel functional polymorphism, 16974 A/C, in the interleukin-12-3' untranslated region is associated with risk of glioma.

Genetic factors are important in the development of glioma. Interleukin-12 (IL-12) is a multifunctional cytokine that induces Interferon (IFN)-gamma secretion and plays an important role in antitumor immunity. Interleukin-27 (IL-27) is a novel IL-12 family member, and the present studies demonstrate that IL-27 mediates a potent antitumor activity. The aim of this study was to investigate whether IL-12 and IL-27 gene polymorphisms and their serum levels are associated with glioma. We analyzed IL-12 gene 16974 A/C and IL-27 gene -964 A/G, 2905 T/G, and 4730 T/C polymorphisms in 210 patients with glioma and 220 matched controls, using polymerase chain reaction-restriction fragment length polymorphism method and DNA sequencing methods, while serum IL-12p40 and IL-27p28 levels were measured by enzyme-linked immunosorbent assay. Serum IL-12p40 and IL-27p28 levels were decreased in patients with glioma compared with controls (p < 0.01). There were significant differences in the genotype and allele frequencies of the IL-12 gene 16974 A/C polymorphism between the group of patients with glioma and the control group (p < 0.05). Moreover, genotypes carrying the IL-12 16974 C variant allele were associated with decreased serum IL-12p40 and IL-27p28 levels compared to the homozygous wild-type genotype in patients with glioma. The IL-12 gene 16974 A/C polymorphism may regulate expression of the serum IL-12p40 and IL-27p28, and associate with increased risk of glioma. Thus, genotypes carrying the IL-12 16974 C variant allele had a decreased ability to produce IL-12 and IL-27, which may contribute to glioma susceptibility.

[Association of the integrin gene polymorphisms with ischemic stroke and plasma lipid levels].

OBJECTIVE: To study the association of integrin alpha-2 (ITGA2) gene C807T, integrin beta-3 (ITGB3) gene T176C polymorphisms with ischemic stroke and the effect of the polymorphisms on plasma lipid and lipoprotein levels. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect the integrin genotypes in 265 patients with ischemic stroke and 280 healthy controls. The plasma lipid and lipoprotein levels were measured by routine method. RESULTS: Plasma total cholesterol (TC), triacylglycerol (TG) and low density lipoprotein-cholesterol (LDL-C) in the patients with ischemic stroke were significantly higher than those in the controls (P< 0.05). The distributions of the ITGB3 gene T176C polymorphism were not different between the ischemic stroke group and control group, but the ITGA2 gene C807T polymorphism was significantly different. The relative risk suffering from ischemic stroke of the T allele carrier was 1.455 times as that of the C allele carrier (OR=1.455, 95%CI: 1.134-1.866). The level of plasma lipid in the T allele carriers was significantly higher than that in the C allele carriers (P< 0.05). CONCLUSION: The ITGA2 gene C807T polymorphism was associated with ischemic stroke, the 807 T allele may be a genetic risk factor for ischemic stroke. The ITGA2 gene C807T polymorphism may affect ischemic stroke through plasma lipid and lipoprotein levels.

Platelet-endothelial cell adhesion molecule-1 gene polymorphism and its soluble level are associated with ischemic stroke.

Inflammation, characterized by the recruitment and adhesion of circulating leukocytes by cellular adhesion molecules, plays an important role in the pathogenesis of atherosclerosis. Genetic analyses of platelet-endothelial cell adhesion molecule-1 (PECAM-1), a key adhesion molecule in the progression of atherosclerosis, have provided conflicting results regarding the role of variation within the PECAM-1 gene and risk for coronary heart disease. No studies have examined the association of this polymorphism with ischemic stroke. Therefore, we investigated that PECAM-1 gene polymorphism and its soluble level are associated with ischemic stroke in Chinese population. We analyzed single-nucleotide polymorphisms of PECAM-1 gene Leu125Val, Asn563Ser, and Gly670Arg in 265 patients with ischemic stroke and 280 age- and sex-matched controls, using polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing method, while soluble PECAM-1 (sPECAM-1) levels were measured by enzyme-linked immunosorbent assay. There were significant differences in the genotype and allele frequencies of PECAM-1 gene Leu125Val polymorphism between the group of patients with ischemic stroke and the control group (p < 0.05). sPECAM-1 levels were increased in patients with ischemic stroke compared with controls (p < 0.01). Moreover, genotypes carrying the PECAM-1 125Val variant allele were associated with increased PECAM-1 levels compared to the homozygous wild-type genotype in patients with ischemic stroke. The Leu125Val polymorphism of PECAM-1 and its sPECAM-1 levels are associated with ischemic stroke in Chinese population. Our data suggest that the PECAM-1 gene may play a role in the development of ischemic stroke.

Association of the single-nucleotide polymorphism and haplotype of the P-selectin gene with ischemic stroke.

Inflammation has recently proven to be associated with the pathogenesis of atherosclerosis and inflammatory genes are good candidates for the risk of developing atherosclerosis. The early phase of atherosclerosis involves the recruitment of inflammatory cells from the circulation and their transendothelial migration. This process is mainly mediated by cellular adhesion molecules. The adhesion molecule P-selectin may play a role in the pathogenesis of atherosclerosis. Polymorphism of P-selectin gene, which may affect the production level of the adhesion molecule, has been associated with a number of atherosclerotic disease. To test this hypothesis, we investigated the relationship of P-selectin gene polymorphisms and ischemic stroke in a Chinese population. We analyzed single nucleotide polymorphisms of P-selectin gene -2,123 G/C, -1,969 G/A, -1,817 T/C and Thr715Pro in three hundred and five patients with ischemic stroke and 280 age and sex matched controls, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing method. There were no significant differences in the genotype, allele and haplotype frequencies of P-selectin gene polymorphisms between the group of patients with ischemic stroke and the control group. Furthermore, there was no significant association of genotype, allele and haplotype at any of the polymorphism in relation to any subtype of ischemic stroke. We did not observe an association between P-selectin gene polymorphisms and ischemic stroke or any subtype of ischemic stroke. However, further studies are needed to explore the complex interaction between environmental factors and P-selectin gene polymorphisms in the risk of ischemic stroke, particularly in ethnically different populations.

[Effection of observation Xinnaoxin capsules in treatment of chronic cerebral circulatory insufficiency].

OBJECTIVE: To observe the therapeutic effect of Xinnaoxin capsules in patients with chronic cerebral circulatory insufficiency. METHOD: Patients with chronic cerebral circulatory insufficiency were divided randomly into two groups: a Xinnaoxin capsules group (n = 60, treated by Xinnaoxin capsules for four 4 weeks), a control group (n = 58, treated by Nimodiping for four weeks). The transcranial doppler (TCD) was used to determined mean velocity (Vm) and auto-viscometer measured hemorheological indices before and after being treated. RESULT: After 4 weeks treatment, the hemorheological indices and mean velocity were obviously improve in Xinnaoxin capsules group (P <0.05), there is significant difference between the effective rate of two groups (88.3%, 70.7%). CONCLUSION: Our study suggest that Xinnaoxin capsules have therapeutic function on chronic cerebral circulatory insufficiency.

[Effect of Panax notoginseng saponins on serum neuronal specific enolase and rehabilitation in patients with cerebral hemorrhage].

OBJECTIVE: To observe the effect of Panax notoginseng saponins (PNS) on serum content of neuronal specific enolase (NSE) and function recovery in patients with cerebral hemorrhage (CH) for exploring the therapeutic action of PNS in treating CH. METHODS: Fifty CH patients with their course of disease not more than 5 days were randomly assigned to two groups, 27 in the PNS group and 23 in the control group, all were treated with conventional treatment, while PNS was given additionally to the PNS group. The serum levels of NSE before and after treatment were determined by electrochemiluminescence, and the recovery of patients, including their neuro-function deficit and daily living activity, was assessed according to scoring by the National Institute of Health Stroke Scale (NIHSS) and Barthel Index (BI) respectively. RESULTS: Before treatment, the difference between the two groups in serum NSE content and scores of NIHSS and BI was insignificant (P > 0.05). However, after 3 weeks of treatment, the level of NSE and score of NIHSS were significantly lower, while score of BI was significantly higher in the PNS group than those in the control group respectively (all P < 0.01). In the PNS group, the level of NSE showed a positive correlation with the score of NIHSS (r = 0.757, P < 0.05), and a negative correlation with the score of BI (r = - 0.803, P < 0.05). CONCLUSION: PNS can effectively protect the neuron and promote functional rehabilitation in patients after CH.

[Effects of Angelica sinensis treatment on the expression of flt-1 and flk-1 mRNA after the ischemic brain injury in rats].

AIM: To investigate the effects of Angelica sinensis on the expression of Flt-1, Flk-1 mRNA after the ischemic brain injury in rats. METHODS: Wistar rats randomly divided into two groups: group A rats underwent middle cerebral artery occlusion (MCAO) for 2 hours by suture, group B rats underwent MCAO for 2 hours meanwhile received treatment with Angelica sinensis (5g/kg). At 1 st d, 3 rd d and 7 th d after reperfusion, 36 rats( n = 18 in each group) were assessed by neurological scale and brain tissue was taken to assess the lesion ration with 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The other rats (n = 3 at different time points in each group) were decapitated at 3 h, 6 h, 12 h , 1 st d, 3 rd d, 7 th d after reperfusion. Quantitative reverse transcription and polymerase chain reaction (RT-PCR) technique was used to examine the gene expression of Flt-1, Flk-1. RESULTS: The neurologic deficit score of rats in group B decreased significantly compared with group A at the same time point (P < 0.05). The infarct volume of group A was significant greater than group B at the same time point after reperfusion (P < 0.01). The results of RT-PCR revealed that the gene expression of Flt-1, Flk-1 in the two groups increased from 3 h after reperfusion and reached its peak at the time of 3 rd d after reperfusion, then declined gradually. The gene expression of Flt-1, Flk-1 in the group B was significantly increased than group A at the same time point (P < 0.01). The gene expression of Flk-1 was positive correlated with Flt-1 in two groups (r = 0.957). CONCLUSION: The increasing amount of Flt-1, Flk-1 expression was enhanced by Angelica sinensis following transient interruption of cerebral blood flow in rats.

[Intercellular adhesion molecule-1 gene K469E polymorphism and genetic susceptibility of ischemic stroke in Chinese Zhuang populations].

OBJECTIVE: To examine the relationship between intercellular adhesion molecule-1 (ICAM-1) gene polymorphism and ischemic stroke (IS) in Chinese Zhuang populations. METHODS: The K469E polymorphism in the exon 6 of ICAM-1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism analysis and DNA sequencing in 205 patients with IS of Zhuang nationality and in 210 healthy controls, and the serum level of ICAM-1 was determined by enzyme-linked immunosorbent assay. RESULTS: The IS group showed significantly higher serum levels of ICAM-1 than did the control group (P < 0.01). There was significant difference in frequencies of allele and genotype in K469E polymorphism between IS and control groups, respectively (P < 0.05). The K allele carriers had 1.424 times the risk of suffering from IS as compared with the E allele carriers (OR = 1.424, 95% CI: 1.071 - 1.894); the serum ICAM-1 level of E allele carriers was significantly higher than that of K allele carriers (501.24 +/- 139.56 ng/ml vs 475.17 +/- 118.35 ng/ml, P < 0.01). CONCLUSION: There is an association between ICAM-1 gene K469E polymorphism and IS, and E allele may be a genetic risk factor of IS among Guangxi Zhuangs, in which the ICAM-1 E allele carriers may have up-regulated expression of ICAM-1 and hence are at a higher risk of ischemic stroke.