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Abstract
This work used a carrageenan-based thrombosis model to determine the preventative effects of Lactobacillus plantarum YS1 (LPYS1) on thrombus. In thrombotic mice, LPYS1 improved the activated partial thromboplastin time (APTT), while decreasing the thrombin time (TT), prothrombin time (PT), and fibrinogen (FIB) content. In thrombotic mouse serum, LPYS1 decreased the levels of malondialdehyde (MDA), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), nuclear factor kappa-B (NF-κB), and interleukin-1 beta (IL-1β), while also increasing the activities of superoxide dismutase (SOD) and catalase (CAT). Moreover, LPYS1 upregulated the mRNA expression levels of copper/zinc-SOD (Cu/Zn-SOD), manganese-SOD (Mn-SOD), and CAT in the colon tissues of thrombotic mice, while downregulating those of NF-κB p65, IL-6, TNF-α, and interferon-gamma (IFN-γ) mRNA. In tail vein vascular tissues, LPYS1 suppressed the mRNA expression levels of NF-κB p65, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. The abundances of both beneficial and pathogenic bacteria were altered by LPYS1. These findings show that LPYS1 has the capacity to protect mice from thrombosis, while also revealing some of the underlying mechanisms of this effect.
Abstract
Nattokinase (NK) is effective in the prevention and treatment of cardiovascular disease. Cucumber is rich in nutrients with low sugar content and is safe for consumption. The aim of this study was to construct a therapeutic cucumber that can express NK, which can prevent and alleviate cardiovascular diseases by consumption. Because the Bitter fruit (Bt) gene contributes to bitter taste but has no obvious effect on the growth and development of cucumber, so the NK-producing cucumber was constructed by replacing the Bt gene with NK by using CRISPR/Cas9. The pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were constructed and transformed into Agrobacterium tumefaciens EHA105, which was transformed into cucumber by floral dip method. The crude extract of NK-producing cucumber had significant thrombolytic activity in vitro. In addition, treatment with the crude extract significantly delayed thrombus tail appearance, and the thrombin time of mice was much longer than that of normal mice. The degrees of coagulation and blood viscosity as well as hemorheological properties improved significantly after crude extract treatment. These findings show that NK-producing cucumber can effectively alleviate thrombosis and improve blood biochemical parameters, providing a new direction for diet therapy against cardiovascular diseases.