Objective To study the effect of silencing the NOD-like receptor family, pyrin domain containing protein 3 (NLRP3) gene on the production of inflammatory factors induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP) in rat brain microvascular endothelial cells (BMECs), and whether NLRP3 inflammasome signaling pathway plays a role in the BMEC model of cerebral small vessel disease induced by proinflammatory agents. Methods BMECs from male Wistar rats were extracted in vitro and the morphology and purity of endothelial cells were identified. BMECs in normal culture were divided into blank control group and LPS+ATP group. The expression levels of NLRP3 inflammasome and downstream inflammatory factor Caspase-1 were detected by Western blot and real-time polymerase chain reaction, and compared by student’s t test between the two groups. Small interfering RNA (siRNA) was used to silence the specific gene NLRP3 in BMECs. After transfection of siRNA NLRP3 and siRNA plasmid negative control into BMECs, the transfected cells were divided into four groups, namely, siNC group (non silenced target gene), siNLRP3 group (silenced target gene), siNC+LPS+ATP group (non silenced target gene and added proinflammatory agents) and siNLRP3+LPS+ATP group (silenced target gene and added proinflammatory agents). The expression levels of NLRP3 and Caspase-1 were detected by Western blot and real-time polymerase chain reaction, and analyzed by analysis of variance for 2-factor factorial design. Results The microvascular segments of rat BMECs were “beaded” after 24 h of isolation and culture; after 48 h, “island” cell clusters were formed; after 72 h, “paving stone” like monolayer cells adhered to the wall and grew. After that, the cells gradually became dense and reached the convergence degree of 80%. The positive rate of BMECs detected by immunofluorescence staining was 96%. In the normally cultured cells, the protein and mRNA expression levels of NLRP3 and Caspase-1 in the LPS+ATP group were higher than those in the blank control group (P<0.05). In the RNA interference cultured cells, the protein and mRNA expression levels of NLRP3 and Caspase-1 in the siNLRP3 group were lower than those in the siNC group, and those expression levels in the siNLRP3+LPS+ATP group were lower than those in the siNC+LPS+ATP group (P<0.05); the protein and mRNA expression levels of NLRP3 and Caspase-1 in the siNC+LPS+ATP group were higher than those in the siNC group, and those expression levels in the siNLRP3+LPS+ATP group were higher than those in the siNLRP3 group (P<0.05). Plasmid transfection and proinflammatory agents intervention had statistically significant interaction effect on the mRNA expression of NLRP3 and Caspase-1 (P<0.05). Conclusions LPS and ATP can promote the release of NLRP3 and Caspase-1 in BMECs. Silencing NLRP3 gene expression can reduce the induction of proinflammatory agents. NLRP3 inflammasome signaling pathway may play a role in the cerebral small vessel disease cell model of rat BMECs induced by proinflammatory agents.
Objective To investigate the influencing factors of moderate to severe disability in migraine patients. Methods Patients diagnosed with migraine between September 2022 and January 2024 in the outpatient service or inpatient Department of Neurology of Baotou Central Hospital and Baotou Eighth Hospital were included. According to the scores of the Migraine Disability Assessment questionnaire, patients were divided into a group with no or mild disabilities and a group with moderate to severe disabilities. The sociodemographic, disease characteristics, and scale datas of two groups of patients were collected, and a multivariate logistic regression model was used to explore the influencing factors of moderate to severe disability in migraine patients. Results A total of 116 patients were included. Among them, there were 49 cases in the group with no or mild disabilities, and 67 cases in the group with moderate to severe disabilities. There were statistically significant differences in gender, duration of headache, severity of headache, number of headache days per month, drug overuse, the scores of Generalized Anxiety Disorder-7 scale, the scores of Patient Health Questionnaire-9 scale, the scores of Pittsburgh Sleep Quality Index scale, the scores of Headache Impact Test scale, the scores of Montreal Cognitive Assessment scale, and the scores of 36-item Short-Form Health Survey questionnaires between the two groups (P<0.05). There was no statistically significant difference in other sociodemographic information and disease characteristics between the two groups of patients (P>0.05). The results of multivariate logistic regression analysis showed that the number of headache days per month, the scores of Generalized Anxiety Disorder-7 scale, the scores of Headache Impact Test scale, and the scores of 36-item Short-Form Health Survey questionnaire were independent influencing factors for moderate to severe disability in migraine patients (P<0.05). Conclusions Headache duration, anxiety disorders and health-related quality of life are influencing factors for moderate to severe disability in migraine patients. Early screening and intervention of influencing factors for migraine patients should be emphasized.