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[腰交感神经节射频热凝对糖尿病性神经病理性疼痛大鼠小胶质细胞活化的影响]

[The influence of lumbar sympathetic ganglion radiofrequency thermocoagulation on the activation of microglia in rats with diabetic neuropathic pain].

作者信息

Zhang J H, Yang C X, Zhong J Y, Zhang L, Xiong Q M, Wang J, Wang H B

机构信息

Department of Anesthesiology, the First People's Hospital of Foshan, Foshan 528000, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2016 Jun 28;96(24):1934-8. doi: 10.3760/cma.j.issn.0376-2491.2016.24.014.

Abstract

OBJECTIVE

To observe the influence of lumbar sympathetic ganglion radiofrequency thermocoagulation on the activation of spinal microglia in rats with diabetic neuropathic pain (DNP).

METHODS

Thirty-six painful diabetic Sprague-Dawley rats induced by 60 mg/kg streptozotocin (STZ) intraperitoneal injection were randomly divided into diabetic neuropathic pain group (group DNP, n=12), Sham operation group (group Sham, n=12) and radiofrequency thermocoagulation group (group R, n=12). Meanwhile another 12 age-matched rats were allocated as normal control group (group N), rats in group N received intraperitoneal injection of equal volume of normal saline. Twenty-eight days after STZ injection, rats in group R received L3 lumbar sympathetic ganglia radiofrequency thermocoagulation on the left side under X-ray guideline after anesthesia with damage time 60 s and damage temperature 60 ℃. Rats in group Sham received puncture positioning, but not thermocoagulation therapy. The mechanical paw withdrawal threshold (PWT) were performed before STZ injection, 7, 14, 21, 28 days after STZ injection and 1, 3, 5, 7, 14 days after radiofrequency thermocoagulation, respectively. Blood glucose were performed before STZ injection, 3, 28 days after STZ injection and 1, 14days after radiofrequency thermocoagulation. After the final behavioral testing, L3-L5 spinal cord tissues were removed to exam the expression of microglia marker OX42 by Western blotting and immunofluorescence technique, and the changes in the expression of inflammation factor IL-1β, IL-6, TNF-α were detected by ELISA technique.

RESULTS

Compared with group N, after 14, 21, 28 days of STZ injection and 1, 3, 5, 7, 14 days of radiofrequency thermocoagulation, the PWT of group DNP and group Sham decreased significantly (P<0.05); Before radiofrequency thermocoagulation, the PWT of rats in group DNP was (3.84±0.83) g, the PWT of rats in group R was (4.45±0.88) g, there was no statistically significant difference between group DNP and group R (t=1.514, P>0.05), but after radiofrequency thermocoagulation, compared with DNP group, the PWT of rats in group R increased significantly (P<0.05), and lasted to 14 d after radiofrequency thermocoagulation. The ratio of spinal microglia marker OX42 and GAPDH, the expression of inflammation factor IL-1β, IL-6, and TNF-α in group N were 0.074±0.023, (35.93±6.16) pg/ml, (92.11±13.23) pg/ml, and (169.50±22.64) pg/ml, respectively. The ratio of spinal microglia marker OX42 and GAPDH, the expression of inflammation factor IL-1β, IL-6, and TNF-α in group DNP were 1.023±0.185, (73.82±9.25) pg/ml, (155.33±21.82) pg/ml, and (298.30±33.21) pg/ml, respectively. The ratio of spinal microglia marker OX42 and GAPDH, the expression of inflammation factor IL-1β, IL-6, and TNF-α in group Sham were 0.951±0.103, (73.00±7.54) pg/ml, (151.02±24.26) pg/ml, and (294.01±36.37) pg/ml, respectively. The ratio of spinal microglia marker OX42 and GAPDH, the expression of inflammation factor IL-1β, IL-6, and TNF-α in group R were 0.563±0.019, (51.81±7.36) pg/ml, (123.24±16.13) pg/ml, and (229.23±29.16) pg/ml, respectively. Compared with group N, the expression of spinal microglia marker OX42 and inflammation factor IL-1β, IL-6, and TNF-α in group DNP, group Sham and group R increased significantly (F=7.501, 348.698, 568.021, 145.110, all P<0.05). Compared with DNP group, the expression of spinal microglia marker OX42 and inflammation factor IL-1β, IL-6, and TNF-α of group R reduced significantly (all P<0.05).

CONCLUSION

The lumbar sympathetic ganglion radiofrequency thermocoagulation can alleviate diabetic neuropathic pain. The mechanism may relate with the inhibition of spinal microglia activation and the lower expression of inflammation factor.

摘要

目的

观察腰交感神经节射频热凝术对糖尿病性神经病理性疼痛(DNP)大鼠脊髓小胶质细胞激活的影响。

方法

将36只腹腔注射60mg/kg链脲佐菌素(STZ)诱导的疼痛性糖尿病Sprague-Dawley大鼠随机分为糖尿病性神经病理性疼痛组(DNP组,n=12)、假手术组(Sham组,n=12)和射频热凝组(R组,n=12)。另取12只年龄匹配的大鼠作为正常对照组(N组),N组大鼠腹腔注射等体积的生理盐水。STZ注射28天后,R组大鼠在麻醉后于X线引导下对左侧L3腰交感神经节进行射频热凝,损伤时间60s,损伤温度60℃。Sham组大鼠进行穿刺定位,但不进行热凝治疗。分别在STZ注射前、注射后7、14、21、28天以及射频热凝后1、3、5、7、14天测定机械缩爪阈值(PWT)。在STZ注射前、注射后3、28天以及射频热凝后1、14天测定血糖。末次行为学检测后,取L3-L5脊髓组织,采用蛋白质免疫印迹法和免疫荧光技术检测小胶质细胞标志物OX42的表达,采用酶联免疫吸附测定(ELISA)技术检测炎症因子IL-1β、IL-6、TNF-α表达的变化。

结果

与N组相比,STZ注射后14、21、28天以及射频热凝后1、3、5、7、14天,DNP组和Sham组的PWT显著降低(P<0.05);射频热凝前,DNP组大鼠的PWT为(3.84±0.83)g,R组大鼠的PWT为(4.45±0.88)g,DNP组与R组之间差异无统计学意义(t=1.514,P>0.05),但射频热凝后,与DNP组相比,R组大鼠的PWT显著升高(P<0.05),并持续至射频热凝后14天。N组脊髓小胶质细胞标志物OX4²与甘油醛-3-磷酸脱氢酶(GAPDH)的比值、炎症因子IL-1β、IL-6和TNF-α的表达分别为0.074±0.023、(35.93±6.16)pg/ml、(92.11±13.23)pg/ml和(169.50±22.64)pg/ml。DNP组脊髓小胶质细胞标志物OX42与GAPDH的比值、炎症因子IL-1β、IL-6和TNF-α的表达分别为1.023±0.185、(73.82±9.25)pg/ml、(155.33±21.82)pg/ml和(298.30±33.21)pg/ml。Sham组脊髓小胶质细胞标志物OX42与GAPDH的比值、炎症因子IL-1β、IL-6和TNF-α的表达分别为0.951±$0.103、(73.00±7.54)pg/ml、(151.02±24.26)pg/ml和(294.01±36.37)pg/ml。R组脊髓小胶质细胞标志物OX42与GAPDH的比值、炎症因子IL-1β、IL-6和TNF-α的表达分别为0.563±0.019、(51.81±7.36)pg/ml、(123.24±16.13)pg/ml和(229.23±29.16)pg/ml。与N组相比,DNP组、Sham组和R组脊髓小胶质细胞标志物OX42以及炎症因子IL-1β、IL-6和TNF-α的表达均显著升高(F=7.501、348.698、568.021、145.110,均P<0.05)。与DNP组相比,R组脊髓小胶质细胞标志物OX42以及炎症因子IL-1β、IL-6和TNF-α的表达均显著降低(均P<0.⁰5)。

结论

腰交感神经节射频热凝术可缓解糖尿病性神经病理性疼痛。其机制可能与抑制脊髓小胶质细胞激活及炎症因子表达降低有关。

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