PDF(Pubmed)
Abstract：
UNASSIGNED: Uropathogenic Escherichia coli (UPEC) activates innate immune response upon invading the urinary tract, whereas UPEC can also enter bladder epithelial cells (BECs) through interactions with fusiform vesicles on cell surfaces and subsequently escape from the vesicles into the cytoplasm to establish intracellular bacterial communities, finally evading the host immune system and leading to recurrent urinary tract infection (RUTI). Tailin Fang II (TLF-II) is a Chinese herbal formulation composed of botanicals that has been clinically proven to be effective in treating urinary tract infection (UTI). However, the underlying therapeutic mechanisms remain poorly understood.
UNASSIGNED: Network pharmacology analysis of TLF-II was conducted. Female Balb/C mice were transurethrally inoculated with UPEC CFT073 strain to establish the UTI mouse model. Levofloxacin was used as a positive control. Mice were randomly divided into four groups: negative control, UTI, TLF-II, and levofloxacin. Histopathological changes in bladder tissues were assessed by evaluating the bladder organ index and performing hematoxylin-eosin staining. The bacterial load in the bladder tissue and urine sample of mice was quantified. Activation of the TLR4-NF-κB pathway was investigated through immunohistochemistry and western blotting. The urinary levels of interleukin (IL)-1β and IL-6 and urine leukocyte counts were monitored. We also determined the protein expressions of markers associated with fusiform vesicles, Rab27b and Galectin-3, and levels of the phosphate transporter protein SLC20A1. Subsequently, the co-localization of Rab27b and SLC20A1 with CFT073 was examined using confocal fluorescence microscopy.
UNASSIGNED: Data of network pharmacology analysis suggested that TLF-II could against UTI through multiple targets and pathways associated with innate immunity and inflammation. Additionally, TLF-II significantly attenuated UPEC-induced bladder injury and reduced the bladder bacterial load. Meanwhile, TLF-II inhibited the expression of TLR4 and NF-κB on BECs and decreased the urine levels of IL-1β and IL-6 and urine leukocyte counts. TLF-II reduced SLC20A1 and Galectin-3 expressions and increased Rab27b expression. The co-localization of SLC20A1 and Rab27b with CFT073 was significantly reduced in the TLF-II group.
UNASSIGNED: Collectively, innate immunity and bacterial escape from fusiform vesicles play important roles in UPEC-induced bladder infections. Our findings suggest that TLF-II combats UPEC-induced bladder infections by effectively mitigating bladder inflammation and preventing bacterial escape from fusiform vesicles into the cytoplasm. The findings suggest that TLF-II is a promising option for treating UTI and reducing its recurrence.
UNASSIGNED: Network pharmacology analysis of TLF-II was conducted. Female Balb/C mice were transurethrally inoculated with UPEC CFT073 strain to establish the UTI mouse model. Levofloxacin was used as a positive control. Mice were randomly divided into four groups: negative control, UTI, TLF-II, and levofloxacin. Histopathological changes in bladder tissues were assessed by evaluating the bladder organ index and performing hematoxylin-eosin staining. The bacterial load in the bladder tissue and urine sample of mice was quantified. Activation of the TLR4-NF-κB pathway was investigated through immunohistochemistry and western blotting. The urinary levels of interleukin (IL)-1β and IL-6 and urine leukocyte counts were monitored. We also determined the protein expressions of markers associated with fusiform vesicles, Rab27b and Galectin-3, and levels of the phosphate transporter protein SLC20A1. Subsequently, the co-localization of Rab27b and SLC20A1 with CFT073 was examined using confocal fluorescence microscopy.
UNASSIGNED: Data of network pharmacology analysis suggested that TLF-II could against UTI through multiple targets and pathways associated with innate immunity and inflammation. Additionally, TLF-II significantly attenuated UPEC-induced bladder injury and reduced the bladder bacterial load. Meanwhile, TLF-II inhibited the expression of TLR4 and NF-κB on BECs and decreased the urine levels of IL-1β and IL-6 and urine leukocyte counts. TLF-II reduced SLC20A1 and Galectin-3 expressions and increased Rab27b expression. The co-localization of SLC20A1 and Rab27b with CFT073 was significantly reduced in the TLF-II group.
UNASSIGNED: Collectively, innate immunity and bacterial escape from fusiform vesicles play important roles in UPEC-induced bladder infections. Our findings suggest that TLF-II combats UPEC-induced bladder infections by effectively mitigating bladder inflammation and preventing bacterial escape from fusiform vesicles into the cytoplasm. The findings suggest that TLF-II is a promising option for treating UTI and reducing its recurrence.
摘要：
■泌尿致病性大肠杆菌（UPEC）在入侵泌尿道时激活先天免疫反应，而UPEC也可以通过与细胞表面梭形囊泡的相互作用进入膀胱上皮细胞（BECs），并随后从囊泡逃逸到细胞质中建立细胞内细菌群落，最终逃避宿主免疫系统并导致复发性尿路感染（RUTI）。TailinFangII（TLF-II）是由植物药组成的中草药制剂，已被临床证明可有效治疗尿路感染（UTI）。然而,潜在的治疗机制仍然知之甚少。
■进行TLF-II的网络药理学分析。雌性Balb/C小鼠经尿道接种UPECCFT073株，建立UTI小鼠模型。左氧氟沙星用作阳性对照。小鼠随机分为四组:阴性对照,UTI,TLF-II,和左氧氟沙星.通过评估膀胱器官指数并进行苏木精-伊红染色来评估膀胱组织的组织病理学变化。定量小鼠的膀胱组织和尿液样品中的细菌负荷。通过免疫组织化学和蛋白质印迹研究TLR4-NF-κB途径的激活。监测尿白细胞介素（IL）-1β和IL-6水平以及尿白细胞计数。我们还确定了与梭形囊泡相关的标志物的蛋白质表达，Rab27b和Galectin-3，以及磷酸盐转运蛋白SLC20A1的水平。随后，使用共聚焦荧光显微镜检查Rab27b和SLC20A1与CFT073的共定位。
■网络药理学分析的数据表明，TLF-II可以通过与先天免疫和炎症相关的多个靶点和途径对抗UTI。此外,TLF-II可显着减轻UPEC诱导的膀胱损伤并减少膀胱细菌负荷。同时,TLF-II抑制BECs上TLR4和NF-κB的表达，降低尿IL-1β和IL-6水平和尿白细胞计数。TLF-II降低SLC20A1和Galectin-3的表达并增加Rab27b的表达。在TLF-II组中,SLC20A1和Rab27b与CFT073的共定位显著减少。
■集体，先天免疫和梭状囊泡的细菌逃逸在UPEC诱导的膀胱感染中起重要作用。我们的发现表明，TLF-II通过有效减轻膀胱炎症和防止细菌从梭形囊泡逃逸到细胞质中来对抗UPEC诱导的膀胱感染。研究结果表明，TLF-II是治疗UTI和减少其复发的有希望的选择。
■进行TLF-II的网络药理学分析。雌性Balb/C小鼠经尿道接种UPECCFT073株，建立UTI小鼠模型。左氧氟沙星用作阳性对照。小鼠随机分为四组:阴性对照,UTI,TLF-II,和左氧氟沙星.通过评估膀胱器官指数并进行苏木精-伊红染色来评估膀胱组织的组织病理学变化。定量小鼠的膀胱组织和尿液样品中的细菌负荷。通过免疫组织化学和蛋白质印迹研究TLR4-NF-κB途径的激活。监测尿白细胞介素（IL）-1β和IL-6水平以及尿白细胞计数。我们还确定了与梭形囊泡相关的标志物的蛋白质表达，Rab27b和Galectin-3，以及磷酸盐转运蛋白SLC20A1的水平。随后，使用共聚焦荧光显微镜检查Rab27b和SLC20A1与CFT073的共定位。
■网络药理学分析的数据表明，TLF-II可以通过与先天免疫和炎症相关的多个靶点和途径对抗UTI。此外,TLF-II可显着减轻UPEC诱导的膀胱损伤并减少膀胱细菌负荷。同时,TLF-II抑制BECs上TLR4和NF-κB的表达，降低尿IL-1β和IL-6水平和尿白细胞计数。TLF-II降低SLC20A1和Galectin-3的表达并增加Rab27b的表达。在TLF-II组中,SLC20A1和Rab27b与CFT073的共定位显著减少。
■集体，先天免疫和梭状囊泡的细菌逃逸在UPEC诱导的膀胱感染中起重要作用。我们的发现表明，TLF-II通过有效减轻膀胱炎症和防止细菌从梭形囊泡逃逸到细胞质中来对抗UPEC诱导的膀胱感染。研究结果表明，TLF-II是治疗UTI和减少其复发的有希望的选择。
