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在大鼠模型中,电针通过激活NGF/TrkA信号通路改善神经源性膀胱功能障碍。

Electroacupuncture improves neurogenic bladder dysfunction through activation of NGF/TrkA signaling in a rat model.

作者信息

Zhang Tong, Yu Jiafeng, Huang Zongqiang, Wang Gang, Zhang Runhe

机构信息

Department of Rehabilitation, First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.

出版信息

J Cell Biochem. 2019 Jun;120(6):9900-9905. doi: 10.1002/jcb.28272. Epub 2018 Dec 28.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture on the morphological change of the bladder tissue and the protein expression levels of NGF, TrkA, p-TrkA, AKT, and p-AKT in the bladder tissue of rats with neurogenic bladder after suprasacral spinal cord injury and to preliminarily explore its partial mechanism of action.

METHODS

Eighty female Sprague-Dawley rats were randomly divided into blank group, model group, electroacupuncture group, model/siNGF group, and electroacupuncture/siNGF group according to random number table method with 16 rats in each group. Eighty Neurogenic bladder models after suprasacral spinal cord injury were established by adopting a modified spinal cord transection method. Electroacupuncture intervention was conducted on the 19th day after modeling. The bladder function was detected by bladder weight, urine output, serum BUN, and urine protein. After treatment for 7 consecutive days, the rats were killed and the bladder tissues were removed rapidly for microscopic observation of morphological change after hematoxylin and eosin stain and for determination of the protein expression levels of NGF, TrkA, p-TrkA, AKT, and p-AKT via Western blot analysis. The transcription of NGF was measured by reverse-transcription polymerase chain reaction.

RESULTS

After treatment, compared with the blank group, the bladder weight of model and electroacupuncture groups were significantly increased (P < 0.05). Compared with the model group, the bladder weight of the electroacupuncture group was decreased (P > 0.05). Compared with the blank group, the urine output of the model group was increased ( P < 0.05). Compared with the blank group, the urine output of the electroacupuncture group was increased ( P > 0.05). Compared with the blank group, the serum BUN of the model group was increased ( P < 0.05). Compared with the blank group, the serum BUN of the electroacupuncture group was increased ( P > 0.05). Compared with the blank group, the urine protein of the model group was increased ( P < 0.05). Compared with the blank group, the urine protein of the electroacupuncture group was increased ( P > 0.05). The expression of NGF, p-TrkA, and p-AKT in the model and electroacupuncture groups was obviously higher than that in the blank group ( P < 0.05). The expression of NGF, p-TrkA, and p-AKT in the electroacupuncture group was higher than that in the model group. The expression of TrkA and AKT were unchanged in blank, model, and electroacupuncture groups ( P > 0.05). After tail vein injection with siNGF lentivirus, the expression of NGF in the model/siNGF group and electroacupuncture/siNGF group was significantly decreased ( P < 0.05). And the protein level of p-AKT and p-TrkA was significantly lower than that of the model and electroacupuncture groups ( P < 0.05).

CONCLUSION

Sacral electroacupuncture therapy can improve the expression of both NGF/TrkA signaling and AKT signaling in the local nerve of the damaged spinal cord, inhibit apoptosis of the damaged spinal cord, protect nerve cells, and promote the recovery of the damaged nerve. At the same time, electroacupuncture can promote the coordination of micturition reflex and improve neurogenic bladder function after the spinal cord injury.

摘要

目的

观察电针对骶上脊髓损伤后神经源性膀胱大鼠膀胱组织形态学变化及膀胱组织中神经生长因子(NGF)、酪氨酸激酶受体A(TrkA)、磷酸化酪氨酸激酶受体A(p-TrkA)、蛋白激酶B(AKT)及磷酸化蛋白激酶B(p-AKT)蛋白表达水平的影响,并初步探讨其部分作用机制。

方法

将80只雌性Sprague-Dawley大鼠按随机数字表法随机分为空白组、模型组、电针组、模型/小干扰RNA(siNGF)组、电针/siNGF组,每组16只。采用改良脊髓横断法建立80只骶上脊髓损伤后神经源性膀胱模型。造模后第19天进行电针干预。通过膀胱重量、尿量、血清尿素氮(BUN)及尿蛋白检测膀胱功能。连续治疗7天后处死大鼠,迅速取出膀胱组织,苏木精-伊红染色后显微镜下观察形态学变化,采用蛋白质免疫印迹法检测膀胱组织中NGF、TrkA、p-TrkA、AKT及p-AKT蛋白表达水平。采用逆转录聚合酶链反应检测NGF转录水平。

结果

治疗后,与空白组比较,模型组和电针组膀胱重量显著增加(P<0.05)。与模型组比较,电针组膀胱重量降低(P>0.05)。与空白组比较,模型组尿量增加(P<0.05)。与空白组比较,电针组尿量增加(P>0.05)。与空白组比较,模型组血清BUN升高(P<0.05)。与空白组比较,电针组血清BUN升高(P>0.05)。与空白组比较,模型组尿蛋白增加(P<0.05)。与空白组比较,电针组尿蛋白增加(P>0.05)。模型组和电针组NGF、p-TrkA及p-AKT表达明显高于空白组(P<0.05)。电针组NGF、p-TrkA及p-AKT表达高于模型组。空白组、模型组和电针组TrkA及AKT表达无变化(P>0.05)。尾静脉注射siNGF慢病毒后,模型/siNGF组和电针/siNGF组NGF表达显著降低(P<0.05)。且p-AKT和p-TrkA蛋白水平显著低于模型组和电针组(P<0.05)。

结论

骶部电针治疗可提高损伤脊髓局部神经中NGF/TrkA信号及AKT信号表达,抑制损伤脊髓凋亡,保护神经细胞,促进损伤神经恢复。同时,电针可促进排尿反射协调,改善脊髓损伤后神经源性膀胱功能。

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