[Effect of moxibustion at "oppositely-located points" on neurogenic bladder after spinal cord injury and endoplasmic reticulum stress pathway in rats]
- PMID: 35403401
- DOI: 10.13703/j.0255-2930.20210929-k0003
[Effect of moxibustion at "oppositely-located points" on neurogenic bladder after spinal cord injury and endoplasmic reticulum stress pathway in rats]
Abstract
Objective: To observe the effect of moxibustion at oppositely-located points "Mingmen" (GV 4) and "Shenque" (CV 8) on the motor function of the hind limbs and bladder function in rats with neurogenic bladder after suprasacral spinal cord injury (SCI), so as to explore the effect of this therapy on bladder tissue apoptosis mediated by endoplasmic reticulum stress pathway.
Methods: Twenty-eight female Wistar rats were randomly divided into a sham-operation group (8 rats) and a model establishment group (20 rats). Using the modified Allen's method, the spinal cord of T10 segment was injured to establish a neurogenic bladder model in the model establishment group. Sixteen rats were modeled successfully and then divided into a model group (8 rats) and a moxibustion group (8 rats). In the moxibustion group, 2 h after consciousness regaining from modeling anesthesia, moxibustion was exerted at "Shenque" (CV 8) and "Mingmen" (GV 4), 2 cones at each acupoint in one intervention. The intervention was administered once every two days and 5-time intervention was required totally. After intervention, Basso, Beattie and Bresnahan locomotor rating scale (BBB) score for the motor function of the hind limbs, and the urodynamics indexes (maximum bladder capacity, urine leakage pressure and bladder compliance) were compared among groups. HE staining method was adopted to observe the morphological changes of bladder tissue. With Western blot method and real-time PCR assay, the protein and mRNA expressions of the endoplasmic reticulum stress-related genes (glucose- regulated protein 78 [GRP78], activating transcription factor 4 [ATF4] and cysteinyl aspartate specific proteinase-12 [Caspase-12]) were determined.
Results: The transitional epithelial cells were arranged irregularly, the bladder wall was getting thinner, and the cellular vacuolar degeneration and neutrophil infiltration were found in the model group. Whereas, compared with the model group, in the moxibustion group, the arrangement of transitional epithelial cells was clear and continuous in layers, the cellular vacuolar degeneration was mild and the infiltration presented in a small amount of neutrophil granulocytes. Compared with the sham-operation group, in the model group, the BBB score was reduced (P<0.01), the maximum bladder capacity and bladder compliance were increased (P<0.01), and the protein expression levels of GRP78, ATF4 and Caspase-12, as well as mRNA expressions were all increased (P<0.01). In comparison with the model group, in the moxibustion group, BBB score was increased (P<0.01), the maximum bladder capacity and bladder compliance were decreased (P<0.01), and the protein and mRNA expression levels of GRP78, ATF4 and Caspase-12 were all decreased (P<0.01).
Conclusion: Moxibustion at the "oppositely-located points" improves the urination function, alleviate urine retention in neurogenic bladder rats after spinal cord injury. The underlying mechanism may be related to the down-regulation of the expressions of GRP78, ATF4 and Caspase-12 in the endoplasmic reticulum stress pathway of the bladder tissues, and thus to alleviate the apoptosis of bladder tissue.
目的:观察艾灸相对穴“命门”“神阙”对骶上脊髓损伤(SCI)后神经源性膀胱大鼠后肢运动及膀胱功能的影响,探究其在内质网应激途径介导的膀胱组织凋亡中发挥的作用。方法:将28只雌性Wistar大鼠随机分为假手术组(8只)和造模组(20只)。造模组采用改良Allen′s法损伤大鼠T10段脊髓制备SCI后神经源性膀胱模型,将造模成功的16只大鼠随机分为模型组(8只)和艾灸组(8只)。艾灸组大鼠于造模麻醉清醒后2 h予艾灸“神阙”“命门”干预,每穴每次2壮,隔日1次,共干预5次。比较各组大鼠干预后后肢运动功能(BBB)评分、尿流动力学指标(膀胱最大容量、漏尿点压力、膀胱顺应性),HE染色法观察膀胱组织形态学变化,Western blot法及实时荧光定量PCR法测定膀胱组织内质网应激相关基因[葡萄糖调节蛋白78(GRP78)、激活转录因子4(ATF4)、半胱氨酸天冬氨酸特异性蛋白酶12(Caspase-12)]蛋白及mRNA表达。结果:模型组大鼠膀胱移行上皮细胞排列紊乱,膀胱壁变薄,可见细胞空泡变性及中性粒细胞浸润;艾灸组大鼠膀胱移行上皮细胞较模型组层次清晰、连续,可见轻度细胞空泡变性及少量中性粒细胞浸润。与假手术组比较,模型组大鼠BBB评分降低(P<0.01),膀胱最大容量及膀胱顺应性增高(P<0.01),膀胱组织GRP78、ATF4、Caspase-12蛋白及mRNA表达增高(P<0.01)。与模型组比较,艾灸组大鼠BBB评分增高(P<0.01),膀胱最大容量及膀胱顺应性降低(P<0.01),膀胱组织GRP78、ATF4、Caspase-12蛋白及mRNA表达降低(P<0.01)。结论:艾灸“相对穴”可改善脊髓损伤后神经源性膀胱大鼠的排尿功能,减轻尿潴留,其机制可能与下调膀胱组织内质网应激通路中GRP78、ATF4、Caspase-12的表达,从而减轻膀胱组织细胞凋亡有关。.
Keywords: apoptosis; endoplasmic reticulum stress; moxibustion; neurogenic bladder; oppositely-located points; spinal cord injury.
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