ABSTRACT
Objective:To investigate the effect of intravenous thrombolysis with recombinant tissue plasminogen activator (rt-PA) on sizes and components of thrombi in patients with acute ischemic stroke (AIS) after mechanical thrombectomy (MT).Methods:Ninety-eight AIS patients accepted MT in Department of Neurology, Huaian First Hospital Affiliated to Nanjing Medical University from June 2018 to June 2022 were enrolled; they were divided into bridging therapy group and direct thrombectomy group according to the treatments. Baseline data, prognoses and thrombus features between the 2 groups were compared.Results:No significant differences were observed between the 2 groups in terms of age, gender, previous medical history, baseline scores, sites and types of vascular occlusion, TOAST causative classifications, time frame, MT frequencies and techniques, successful recanalization, or prognoses 90 d after MT ( P>0.05). Compared with the direct thrombectomy group, the bridging therapy group had significantly reduced thrombus area (37[22, 78] mm 2vs. 27[14, 62] mm 2) and areas of major pathological components [red blood cells: 16.1([9.0, 37.4] mm 2vs. 11.8[6.1, 22.1] mm 2, fibrin/platelets: 19.5[9.5, 26.4] mm 2vs. 10.7[5.0, 22.8] mm 2, white blood cells: 4.0[2.2, 8.0] mm 2vs. 2.4[1.4, 5.7] mm 2, P<0.05); however, no significant differences in proportions of areas of major pathological components were noted between the 2 groups of patients ( P>0.05). Conclusion:The rt-PA intravenous thrombolysis can significantly reduce the thrombi volume and decrease the contents of major thrombus components in AIS patients after MT, but not change the proportions of major pathological components in thrombi.
ABSTRACT
Objective:To explore the regulatory mechanism of α-synuclein in the degradation of autophagy-lysosome pathway(ALP) in Parkinson disease(PD) model cells after interference or overexpression of dynein heavy chain(Dynhc) gene.Methods:SH-SY5Y cells were divided into control group, PD group, Dynhc interference group, Dynhc overexpression group, and Dynhc interference+ rapamycin group according to experimental requirements.Using Western blot to detect Dynhc, α-synuclein, microtubule-associated protein l light chain 3 (LC3), lysosome-associated membrane protein 2 (LAMP2), tubulin, dynein activator protein p150, and kinesin KIF5B.Flow cytometry was used to detect the level of cell apoptosis.Immunoconfocal microscopy was used to observe the structure of tubulin and the co-localization of LC3 and LAMP.SPSS 23.0 software was used for statistical analysis.One-way ANOVA was used for inter group comparisons, and further pairwise comparisons were conducted by LSD- t test. Results:There were statistically significant differences in the expression of α-synuclein, autophagy-related proteins, microtubules, and microtubule-related proteins among cells in the 5 groups(all P<0.001). The protein expression levels of Dynhc, α-synuclein, LC3, LAMP2, p150, and KIF5B in the PD group were higher than those in the control group (all P<0.05). The protein levels of Dynhc, LAMP2, tubulin and p150 in the Dynhc interference group were lower than those in the PD group (all P<0.05), while the protein levels of α-synuclein, LC3 and KIF5B were higher than those in the PD group (all P<0.05). The protein levels of α-synuclein, LC3, and KIF5B in the Dynhc overexpression group were lower than those in the PD group (all P<0.05), while the protein levels of Dynhc, LAMP2 and p150 were higher than those in the PD group (all P<0.05). The protein level of LC3 in the Dynhc interference+ rapamycin group was higher than that in the Dynhc interference group ( P<0.05). There were no statistically significant differences in the protein levels of Dynhc, α-synuclein, LAMP2, microtubule protein, p150 and KIF5B compared to the Dynhc interference group (all P>0.05). Compared with the control group, the cell apoptosis rate in PD group increased((12.77±1.66)%, (7.64±1.45)%), the microtubule morphology remained unchanged, and autophagosomes fused more with lysosomes. Compared with the PD group, the cell apoptosis rate of Dynhc overexpression group decreased, and there was no significant change in microtubule structure, and there was more fusion between autophagosomes and lysosomes.Compared with the PD group, the cell apoptosis rat of Dynhc interference group increased((18.45±1.91)%), and the microtubule morphology was sparse, and there was less fusion between autophagosomes and lysosomes. Compared with the PD group, the Dynhc overexpression group showed a decrease in cell apoptosis rate ((9.95±1.56)%), no significant changes in microtubule structure, and more fusion between autophagosomes and lysosomes.Compared with the Dynhc interference group, the Dynhc interference+ rapamycin group showed no significant changes in cell apoptosis rate ((19.05±2.46)%), microtubule morphology, and fusion of autophagosomes and lysosomes. Conclusion:Dynhc can reduce cell apoptosis by enhancing cell ALP function, increasing the degradation of α-synuclein and maintaining of microtubule structure integrity.