电针调控肿瘤坏死因子受体1/受体相互作用蛋白激酶1/核因子κB轴对骶髓损伤后神经源性膀胱尿潴留大鼠尿动力学的影响

Electroacupuncture improves urodynamics in rats with neurogenic bladder urinary retention after sacral cord injury by regulating TNFR1/RIPK1/NF-κB axis

目的探究电针调控肿瘤坏死因子受体1/受体相互作用蛋白激酶1/核因子κB(TNFR1/RIPK1/NF-κB)轴对骶髓损伤(SCI)后神经源性膀胱(NB)尿潴留大鼠的影响及机制。方法采用随机数字表法将SD大鼠分为空白组、模型组、电针组和电针对照组,每组10只。建立SCI后NB尿潴留模型,电针组取穴“次髎”“中极”“三阴交”,电针对照组取三穴对照点,行电针治疗7 d。各组大鼠治疗前、后行尿动力学检测;取大鼠脊髓组织,实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测TNFR1、RIPK1表达;蛋白质印迹法(Western blotting)检测TNFR1、RIPK1、p-IKKβ/IKKβ、p-NF-κB p65/NF-κB p65和p-IκBα/IκBα蛋白表达;酶联免疫吸附剂测定法(ELISA)检测TNF-α、IL-1β、IL-6、IFN-γ含量。该研究起止时间为2019年1月至2021年12月。结果与空白组比较,模型组大鼠膀胱最大容量[(4.46±0.82)mL比(1.13±0.22)mL,P<0.05]、膀胱基础压[(32.00±4.23)cmH_(2)O比(22.97±3.71)cmH_(2)O,P<0.05]、漏尿点压力[(54.02±7.61)cmH_(2)O比(38.85±5.32)cmH_(2)O,P<0.05]和膀胱顺应性[(0.19±0.03)mL/cmH_(2)O比(0.08±0.01)mL/cmH_(2)O,P<0.05]升高,TNFR1、RIPK1、p-IKKβ/IKKβ、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα表达及TNF-α、IL-1β、IL-6、IFN-γ含量增加(P<0.05);与模型组比较,电针组治疗后大鼠膀胱最大容量[(2.29±0.43)mL比(4.46±0.82)mL,P<0.05]、膀胱基础压[(23.69±3.43)cmH_(2)O比(32.00±4.23)cmH_(2)O,P<0.05]、漏尿点压力[(40.11±5.91)cmH_(2)O比(54.02±7.61)cmH_(2)O,P<0.05]和膀胱顺应性[(0.12±0.02)mL/cmH_(2)O比(0.19±0.03)mL/cmH_(2)O,P<0.05]降低,TNFR1、RIPK1、p-IKKβ/IKKβ、p-NF-κB p65/NF-κB p65、p-IκBα/IκBα表达及TNF-α、IL-1β、IL-6、IFN-γ含量减少(P<0.05);电针对照组与模型组比较差异无统计学意义(P>0.05)。结论电针“次髎”“中极”“三阴交”可改善SCI后NB尿潴留大鼠膀胱功能,机制与抑制TNFR1/RIPK1/NF-κB轴活化,抑制炎症反应有关。

Objective To explore the effect and mechanism of electroacupuncture on rat with neurogenic bladder(NB)urinary retention after sacral cord injury(SCI)by regulating the tumor necrosis factor receptor 1/receptor-interacting protein kinase 1/nuclear factor kappa-B(TNFR1/RIPK1/NF-κB)axis.Methods The SD rats were assigned into blank group,model group,electroacupuncture group and electroacupuncture control group,with 10 rats in each group.A model of NB urine retention after SCI was established.The electroacupuncture group took the points of"Ciliao(BL32)""Zhongji(RN3)",and"Sanyinjiao(SP6)",and the control group took the three-acupoint control points and treated them with electroacupuncture for 7 days.Urodynamic tests were performed before and after treatment in each group of rats.The rat spinal cord tissue was collected,and the expression of TNFR1 and RIPK1 was detected by qRT-PCR,the expression of TNFR1,RIPK1,p-IKKβ/IKKβ,p-NF-κB p65/NF-κB p65 and p-IκBα/IκBαwas detected by Western Blotting,the content of TNF-α,IL-1β,IL-6,IFN-γwas detected by Enzyme-linked immunosorbent assay(ELISA).The study began and ended from January 2019 to December 2021.Results Compared with the blank group,the maximum bladder capacity[(4.46±0.82)mL vs.(1.13±0.22)mL,P<0.05],basal bladder pressure[(32.00±4.23)cmH_(2)O vs.(22.97±3.71)cmH_(2)O,P<0.05],leakage point pressure[(54.02±7.61)cmH_(2)O vs.(38.85±5.32)cmH_(2)O,P<0.05]and bladder compliance[(0.19±0.03)mL/cmH_(2)O vs.(0.08±0.01)mL/cmH_(2)O,P<0.05]increased in the model group,the expression of TNFR1,RIPK1,p-IKKβ/IKKβ,p-NF-κB p65/NF-κB p65,p-IκBα/IκBαand the content of TNF-α,IL-1β,IL-6 and IFN-γincreased,and the difference was statistically significant(P<0.05).Compared with the model group,the maximum bladder capacity[(2.29±0.43)mL vs.(4.46±0.82)mL,P<0.05],basal bladder pressure[(23.69±3.43)cmH_(2)O vs.(32.00±4.23)cmH_(2)O,P<0.05],leakage point pressure[(40.11±5.91)cmH_(2)O vs.(54.02±7.61)cmH_(2)O,P<0.05]and bladder compliance[(0.12±0.02)mL/cmH_(2)O vs.(0.19±0.03)mL/cmH_(2)O,P<0.05]of rats in the electroacupuncture group decreased after treatment,the expression of TNFR1,RIPK1,p-IKKβ/IKKβ,p-NF-κB p65/NF-κB p65,p-IκBα/IκBαand the content of TNF-α,IL-1β,IL-6,IFN-γdecreased(P<0.05).There was no significant difference between the electroacupuncture control group and the model group(P>0.05).Conclusion Electroacupuncture of"Ciliao(BL32)""Zhongji(RN3)",and"Sanyinjiao(SP6)"can improve the bladder function of NB urinary retention rats after SCI.The mechanism is related to inhibiting the activation of TNFR1/RIPK1/NF-κB pathway and inhibiting inflammation.