PDF(Pubmed)
Abstract：
BACKGROUND: Diabetic bladder dysfunction (DBD) is driven in part by inflammation which dysregulates prostaglandin release in the bladder. Precise inflammatory mechanisms responsible for such dysregulation have been elusive. Since prostaglandins impact bladder contractility, elucidating these mechanisms may yield potential therapeutic targets for DBD. In female Type 1 diabetic Akita mice, inflammation mediated by the nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome is responsible for DBD. Here, we utilized female Akita mice crossbred with NLRP3 knock-out mice to determine how NLRP3-driven inflammation impacts prostaglandin release within the bladder and prostaglandin-mediated bladder contractions.
METHODS: Akita mice were crossbred with NLRP3-/- mice to yield four groups of non-diabetics and diabetics with and without the NLRP3 gene. Females were aged to 30 weeks when Akitas typically exhibit DBD. Urothelia and detrusors were stretched ex vivo to release prostaglandins. Prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) were quantified using enzyme linked immunosorbent assays (ELISA). In separate samples, ex vivo contractile force to PGE2 and PGF2α +/- the prostaglandin F (FP) receptor antagonist, AL8810, was measured. FP receptor protein expression was determined via western blotting.
RESULTS: Stretch-induced PGE2 release increases in urothelia but decreases in detrusors of diabetics. However, PGE2-mediated bladder contractions are not impacted. Conversely, diabetics show no changes in PGF2α release, but PGF2α-mediated contractions increase significantly. This is likely due to signaling through the FP receptors as FP receptor antagonism prevents this increase and diabetics demonstrate a four-fold increase in FP receptor proteins. Without NLRP3-mediated inflammation, changes in prostaglandin release, contractility, and receptor expression do not occur.
CONCLUSIONS: NLRP3-dependent inflammation dysregulates prostaglandin release and prostaglandin-mediated bladder contractions in diabetic female Akita mice via FP receptor upregulation.
METHODS: Akita mice were crossbred with NLRP3-/- mice to yield four groups of non-diabetics and diabetics with and without the NLRP3 gene. Females were aged to 30 weeks when Akitas typically exhibit DBD. Urothelia and detrusors were stretched ex vivo to release prostaglandins. Prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) were quantified using enzyme linked immunosorbent assays (ELISA). In separate samples, ex vivo contractile force to PGE2 and PGF2α +/- the prostaglandin F (FP) receptor antagonist, AL8810, was measured. FP receptor protein expression was determined via western blotting.
RESULTS: Stretch-induced PGE2 release increases in urothelia but decreases in detrusors of diabetics. However, PGE2-mediated bladder contractions are not impacted. Conversely, diabetics show no changes in PGF2α release, but PGF2α-mediated contractions increase significantly. This is likely due to signaling through the FP receptors as FP receptor antagonism prevents this increase and diabetics demonstrate a four-fold increase in FP receptor proteins. Without NLRP3-mediated inflammation, changes in prostaglandin release, contractility, and receptor expression do not occur.
CONCLUSIONS: NLRP3-dependent inflammation dysregulates prostaglandin release and prostaglandin-mediated bladder contractions in diabetic female Akita mice via FP receptor upregulation.
摘要：
背景：糖尿病性膀胱功能障碍（DBD）部分是由炎症引起的，该炎症失调了膀胱中前列腺素的释放。导致这种失调的精确炎症机制是难以捉摸的。由于前列腺素影响膀胱收缩性，阐明这些机制可能会产生DBD的潜在治疗靶点。在雌性1型糖尿病秋田小鼠中，由核苷酸结合域介导的炎症，富含亮氨酸的家族，含pyrin结构域-3(NLRP3)炎性体负责DBD。这里,我们利用与NLRP3敲除小鼠杂交的雌性秋田小鼠来确定NLRP3驱动的炎症如何影响膀胱内前列腺素释放和前列腺素介导的膀胱收缩.
方法：秋田小鼠与NLRP3-/-小鼠杂交，得到四组非糖尿病患者和有和没有NLRP3基因的糖尿病患者。当Akitas通常表现出DBD时，雌性的年龄为30周。离体拉伸尿路和逼尿肌以释放前列腺素。使用酶联免疫吸附测定（ELISA）定量前列腺素E2（PGE2）和前列腺素F2α（PGF2α）。在单独的样品中，体外收缩力PGE2和PGF2α+/-前列腺素F（FP）受体拮抗剂，AL8810，进行了测量。通过蛋白质印迹法测定FP受体蛋白表达。
结果：拉伸诱导的PGE2释放在糖尿病患者的尿路中增加，但在逼尿肌中减少。然而,PGE2介导的膀胱收缩不受影响。相反，糖尿病患者的PGF2α释放没有变化，但PGF2α介导的收缩显著增加。这可能是由于通过FP受体的信号传导,因为FP受体拮抗作用阻止了这种增加,并且糖尿病患者表现出FP受体蛋白的四倍增加。没有NLRP3介导的炎症，前列腺素释放的变化，收缩性，和受体表达不发生。
结论：NLRP3依赖性炎症通过FP受体上调调节糖尿病雌性秋田小鼠的前列腺素释放和前列腺素介导的膀胱收缩。
方法：秋田小鼠与NLRP3-
结果：拉伸诱导的PGE2释放在糖尿病患者的尿路中增加，但在逼尿肌中减少。然而,PGE2介导的膀胱收缩不受影响。相反，糖尿病患者的PGF2α释放没有变化，但PGF2α介导的收缩显著增加。这可能是由于通过FP受体的信号传导,因为FP受体拮抗作用阻止了这种增加,并且糖尿病患者表现出FP受体蛋白的四倍增加。没有NLRP3介导的炎症，前列腺素释放的变化，收缩性，和受体表达不发生。
结论：NLRP3依赖性炎症通过FP受体上调调节糖尿病雌性秋田小鼠的前列腺素释放和前列腺素介导的膀胱收缩。
