Zhang Heng, Liu Li-Mei, Lu Gen-Sheng, Xiong En-Qing, Li Wei-Bing, Zhou Zhan-Song, Zhang Jia-Hua, Pan Jin-Hong, Chen Zhi-Wen, Li Long-Kun, Yang Zhong, Song Bo
PLA Center of Urology, Southwest Hospital, Chongqing 400038, China.
Zhonghua Nan Ke Xue. 2009 Nov;15(11):1021-7.
To observe the expressions of the substance P (SP) mRNA and neurokinin-1 receptor (NK-1R) in the posterior horn of the L5 - S2 spinal cord in the rat model of chronic prostatitis pain, and to investigate the changes in the activation of astrocytes and influence of SP on this activation in rat spinal cord astrocytes cultured in vitro.
The rat model of chronic prostatitis pain was established by injection of complete Freund's adjuvant (CFA) and assessed by the tail flick threshold test, the control rats injected with sodium chloride and all observed at 0, 14 and 28 days. Changes in the expressions of SP mRNA, NK-1R, glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) in the posterior horn of the L5 - S2 spinal cord were detected by RT-PCR and Western blot. Rat spinal cord astrocytes were cultured in vitro and divided into a control group, cultured with ITS cell culture fluid, and two experiment groups, with Group 1 stimulated with SP at the concentration of 10(-9) - 10(-6) mol/L for 12 hours followed by determination of the expressions of TNF-alpha, IL-1beta, NO and NOS by ELISA and nitrate reductase and colorimetric methods, and Group 2 at 10(-7) mol/L for 0, 24, 48 and 72 hours followed by detection of the GFAP expression by Western blot.
The expressions of SP mRNA, NK-1 R, GFAP, TNF-alpha and iNOS in the posterior horn of the L5 - S2 spinal cord were obviously higher in the rat prostatitis pain models than in the controls, successively higher at 28 than at 14 and 0 d (P < 0.01), and so was the expression of GFAP at 28 than at 14 d in the experiment groups (P < 0.05). SP induced a gradual increase at 10(-7) mol/L in the expression of GFAP in the spinal cord astrocytes at 0 -72 h, significantly different from that of the control group (P < 0.01), and it promoted the excretion of TNF-alpha and IL-1beta and the activity of NO and NOS at 10(-9) - 10(-6) mol/L at 12 h in a concentration-dependent manner, with marked differences between the experiment and control groups (P < 0.01, P < 0.05). But a decreased excretion of IL-1 beta was observed in the 10(-6) mol/L group, though with no significant difference from the control (P > 0.05).
Chronic prostatitis pain could upregulate the expressions of the excitatory transmitter SP and receptor in the L5 - S2 spinal cord, and result in the activation of astrocytes and increased excretion of proinflammatory cytokines, which may be associated with the persistence and generalization of prostatitis pain.
观察慢性前列腺炎疼痛大鼠模型L5 - S2脊髓后角中P物质(SP)mRNA和神经激肽-1受体(NK-1R)的表达,并研究体外培养的大鼠脊髓星形胶质细胞中星形胶质细胞激活的变化以及SP对这种激活的影响。
通过注射完全弗氏佐剂(CFA)建立慢性前列腺炎疼痛大鼠模型,并通过甩尾阈值试验进行评估,注射氯化钠的对照大鼠在0、14和28天进行观察。采用RT-PCR和蛋白质免疫印迹法检测L5 - S2脊髓后角中SP mRNA、NK-1R、胶质纤维酸性蛋白(GFAP)、肿瘤坏死因子-α(TNF-α)和诱导型一氧化氮合酶(iNOS)的表达变化。体外培养大鼠脊髓星形胶质细胞,分为对照组(用ITS细胞培养液培养)和两个实验组,实验组1用浓度为10(-9) - 10(-6) mol/L的SP刺激12小时,然后用ELISA和硝酸还原酶比色法测定TNF-α、IL-1β、NO和NOS的表达;实验组2用10(-7) mol/L的SP刺激0、24、48和72小时,然后用蛋白质免疫印迹法检测GFAP的表达。
大鼠前列腺炎疼痛模型L5 - S2脊髓后角中SP mRNA、NK-1R、GFAP、TNF-α和iNOS的表达明显高于对照组,在28天时依次高于14天和0天(P < 0.01),实验组中28天时GFAP的表达也高于14天(P < 0.05)。SP在0 - 72小时使脊髓星形胶质细胞中GFAP的表达在10(-7) mol/L时逐渐增加,与对照组有显著差异(P < 0.01),并且在浓度为10(-9) - 10(-6) mol/L时,12小时能以浓度依赖的方式促进TNF-α和IL-1β的分泌以及NO和NOS的活性,实验组与对照组之间有显著差异(P < 0.01,P < 0.05)。但在10(-6) mol/L组中观察到IL-1β的分泌减少,尽管与对照组无显著差异(P > 0.05)。
慢性前列腺炎疼痛可上调L5 - S2脊髓中兴奋性递质SP及其受体的表达,导致星形胶质细胞激活和促炎细胞因子分泌增加,这可能与前列腺炎疼痛的持续和扩散有关。