鞘內(nèi)注射甘珀酸劑量依賴地緩解腰5脊神經(jīng)切斷大鼠的痛覺(jué)高敏

李雪飛,許　倩,王　芬,鄭　曼,劉清珍,李偉彥

(1. 南京中醫(yī)藥大學(xué)附屬醫(yī)院麻醉科，江蘇 南京　210029；2. 南京軍區(qū)總醫(yī)院麻醉科，江蘇 南京　210002)

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鞘內(nèi)注射甘珀酸劑量依賴地緩解腰5脊神經(jīng)切斷大鼠的痛覺(jué)高敏

李雪飛1,許倩1,王芬1,鄭曼1,劉清珍2,李偉彥2

(1. 南京中醫(yī)藥大學(xué)附屬醫(yī)院麻醉科，江蘇 南京210029；2. 南京軍區(qū)總醫(yī)院麻醉科，江蘇 南京210002)

摘要:目的觀察鞘內(nèi)注射甘珀酸(carbenoxolone, CBX)對(duì)腰5脊神經(jīng)切斷(L5 spinal nerve transaction ，SNT)大鼠的鎮(zhèn)痛作用及其相關(guān)機(jī)制。方法60只♂ SD大鼠，隨機(jī)分成5組(n=12)：Ⅰ.假手術(shù)組，Ⅱ.模型組，Ⅲ.SNT+CBX(0.05 μg)，Ⅳ.SNT+CBX(0.5 μg)，Ⅴ.SNT+CBX(5 μg)。Ⅰ組僅暴露腰5脊神經(jīng)，Ⅱ至Ⅴ組行SNT。術(shù)后10 d，Ⅰ和Ⅱ組鞘內(nèi)注射生理鹽水10 μL，Ⅲ至Ⅴ組注射CBX 0.05 μg(10 μL)、0.5 μg(10 μL)和5 μg(10 μL)，分別于術(shù)前1 d，術(shù)后1、3、5、7、10 d各組注射生理鹽水和CBX，前1 h和給藥后1、2、4、6 h測(cè)雙側(cè)機(jī)械痛閾值(mechanical withdrawl thresholds MWT)，免疫組化及ELISA法觀察給藥后2h后脊髓背角膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein, GFAP)和TNF-α、IL-1β表達(dá)的變化。結(jié)果與術(shù)前相比，Ⅱ至Ⅴ組雙側(cè)MWT明顯降低；與給藥前1 h相比，Ⅱ和Ⅲ組給藥后1、2、4、6 h雙側(cè)MWT均無(wú)明顯差異；Ⅳ組損傷側(cè)給藥后1、2、4 h MWT無(wú)明顯差異，而對(duì)側(cè)MWT明顯提高，同時(shí)該側(cè)GFAP和TNF-α、IL-1β的表達(dá)也降低；Ⅴ組給藥后1、2、4 h雙側(cè)MWT明顯提高，雙側(cè)GFAP和TNF-α、IL-1β的表達(dá)也明顯降低。結(jié)論鞘內(nèi)注射CBX可緩解SNT大鼠雙側(cè)MWT，其機(jī)制可能與其抑制脊髓背角星形膠質(zhì)細(xì)胞的活化及TNF-α和IL-1β釋放減少有關(guān)。

關(guān)鍵詞:甘珀酸；神經(jīng)病理性疼痛；痛覺(jué)高敏；脊髓背角；星形膠質(zhì)細(xì)胞； TNF-α；IL-1β

越來(lái)越多的證據(jù)表明，單側(cè)神經(jīng)的損傷會(huì)導(dǎo)致雙側(cè)神經(jīng)的細(xì)胞和分子結(jié)構(gòu)的變化，并導(dǎo)致雙側(cè)痛覺(jué)高敏，把這種現(xiàn)象稱為鏡像痛(mirror image-pain, MIP)，它是神經(jīng)病理性痛(neuropathic pain, NP)中的一種特殊現(xiàn)象[1]。

到目前為止，MIP的發(fā)生機(jī)制尚不清楚，隨著對(duì)膠質(zhì)細(xì)胞研究的不斷深入，膠質(zhì)細(xì)胞在NP中的作用越來(lái)越受到關(guān)注，神經(jīng)的損傷或炎癥可導(dǎo)致膠質(zhì)細(xì)胞的活化及增生，這些膠質(zhì)細(xì)胞會(huì)釋放一些物質(zhì)，反過(guò)來(lái)調(diào)控傷害性神經(jīng)元的興奮性[2]。研究發(fā)現(xiàn)，星形膠質(zhì)細(xì)胞在MIP的發(fā)生中可能扮演著更重要角色。星形膠質(zhì)細(xì)胞之間信號(hào)傳遞的一個(gè)重要機(jī)制是縫隙連接(gap junction, GJ)，損傷側(cè)的傷害性信號(hào)可以通過(guò)GJ以鈣波的形式傳遞到對(duì)側(cè)脊髓，從而促進(jìn)疼痛的傳導(dǎo)[3-4]。

甘珀酸(carbenoxolone, CBX)是一種GJ阻滯劑，使用CBX阻滯GJ可以抑制多種持續(xù)性疼痛動(dòng)物模型中的痛覺(jué)高敏。

本實(shí)驗(yàn)通過(guò)鞘內(nèi)注射不同劑量的CBX來(lái)觀察對(duì)腰5脊神經(jīng)切斷(SNT)大鼠的雙側(cè)機(jī)械痛閾(mechanical withdraw threshold, MWT)的影響，以及雙側(cè)脊髓背角膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein, GFAP)和TNF-α、IL-1β表達(dá)的變化，來(lái)探討星形膠質(zhì)細(xì)胞及炎性介質(zhì)參與大鼠MIP的可能機(jī)制。

1材料與方法

1.1動(dòng)物模型

體質(zhì)量160～180 g的成年♂ Sprague-Dawley大鼠60只(由南京軍區(qū)總醫(yī)院動(dòng)物實(shí)驗(yàn)中心提供，清潔級(jí))，單籠飼養(yǎng)。動(dòng)物飼養(yǎng)室溫為(23±3)℃，周期光照8 ∶00～20 ∶00，大鼠自由進(jìn)食，飲水。所有實(shí)驗(yàn)均在光照期間完成。以2%的戊巴比妥(Sigma公司，美國(guó))50 mg·kg-1腹腔注射麻醉，根據(jù)Kim[5]方法創(chuàng)建模型。

1.2藥物及分組

甘珀酸(Sigma,美國(guó))；兔抗大鼠和膠原纖維酸性蛋白(anti-glial fibrillary acidic protein, GFAP)抗體(Sigma, 美國(guó))；ELISA試劑盒。隨機(jī)分成5組(n=12)，Ⅰ. 假手術(shù)組 (Sham+NS)，Ⅱ.模型組(SNT+NS)，Ⅲ. SNT+CBX(0.05 μg)，Ⅳ. SNT+CBX(0.5 μg)，Ⅴ. SNT+CBX(5 μg)。

1.3鞘內(nèi)注射

參照Mestre等[6]所建立的方法。大鼠麻醉后用連有PE-10導(dǎo)管的稍鈍針頭經(jīng)腰5(L5)和腰4(L4)椎間隙行椎管穿刺，以動(dòng)物出現(xiàn)突然地側(cè)向甩尾運(yùn)動(dòng)，作為穿刺成功的標(biāo)志，鞘內(nèi)注射采用微量進(jìn)樣器進(jìn)行。術(shù)后10 d，Ⅰ組和Ⅱ組鞘內(nèi)注射生理鹽水10μL，Ⅲ至Ⅴ組鞘內(nèi)注射CBX 0.05 μg(10 μL)、0.5 μg(10 μL)和5 μg(10 μL)，CBX濃度的選擇參考既往文獻(xiàn)。

1.4機(jī)械痛閾的測(cè)定

5組大鼠，每組隨機(jī)取6只固定進(jìn)行行為學(xué)測(cè)定，分別于術(shù)前1 d，術(shù)后1 、3、5、7、10 d及給藥后1、2、4、6 h測(cè)定雙側(cè)MWT。MWT測(cè)定：將大鼠分別放置于金屬篩網(wǎng)上的有機(jī)玻璃箱里，安靜15 min，以Von Frey 纖維垂直刺其后肢足底中部皮膚，持續(xù)≤4 s，大鼠出現(xiàn)抬足、舔足、躲避等反應(yīng)時(shí)，讀Electrovon Frey 讀數(shù)器上顯示的最大值(g)，每只大鼠重復(fù)測(cè)量5次，間隔5 min，去除最大和最小值，計(jì)算3 次的平均值即為大鼠的MWT值。

1.5免疫組織化學(xué)染色

5組大鼠術(shù)后10 d鞘內(nèi)注射CBX 2 h后，隨機(jī)取3只按以下方法取材檢測(cè)。2%的戊巴比妥鈉50 mg·kg-1經(jīng)腹腔注射麻醉，開(kāi)胸后經(jīng)左心室插管至升主動(dòng)脈，依次灌注生理鹽水250 mL、4%多聚甲醛(pH 7.4)0.1 mL·L-1、PBS緩沖液250 mL，約1 h后取大鼠脊髓腰膨大，于上述固定液中固定24 h。經(jīng)梯度酒精脫水，二甲苯透明石蠟包埋，進(jìn)行連續(xù)切片，片厚40 μm，每個(gè)蠟塊連續(xù)切5片。以ABC法作GFAP免疫組化染色。染片用半自動(dòng)圖像分析儀進(jìn)行圖像分析，計(jì)算各切片GFAP陽(yáng)性細(xì)胞數(shù)表示星形膠質(zhì)細(xì)胞表達(dá)的強(qiáng)度。胞質(zhì)出現(xiàn)棕黃色顆粒沉積為GFAP免疫組化陽(yáng)性反應(yīng)細(xì)胞。

1.6TNF-α和IL-1β表達(dá)檢測(cè)

ELISA測(cè)定雙側(cè)脊髓背角TNF-α和IL-1β濃度。按照試劑盒說(shuō)明書(shū)操作,并在波長(zhǎng)為450 nm,參考波長(zhǎng)為 620 nm處,用酶標(biāo)儀(美國(guó) Bio-Rad公司)測(cè)定光密度,根據(jù)標(biāo)準(zhǔn)曲線算出TNF-α和IL-1β濃度。

1.7統(tǒng)計(jì)分析

2結(jié)果

2.1MWT測(cè)定結(jié)果

與Ⅰ組相比，Ⅱ至Ⅴ組損傷側(cè)術(shù)后1、3、5、7、10 d MWT均明顯下降(P<0.05)，術(shù)后5 d降至最低，持續(xù)到10 d，而對(duì)側(cè)術(shù)后7 d降至最低，持續(xù)到10 d；與術(shù)后10 d給藥前1 h相比，Ⅱ和Ⅲ組給藥后1、2、4、6 h雙側(cè)MWT均無(wú)明顯差異(P>0.05)；Ⅳ組給藥后1、2、4、6 h損傷側(cè)MWT無(wú)明顯差異(P>0.05)，而對(duì)側(cè)給藥后1 h MWT即開(kāi)始提高(P<0.05)，2 h時(shí)達(dá)高峰(P<0.05)，6 h時(shí)與給藥前無(wú)明顯差異(P>0.05)；Ⅴ組給藥后1、2、4 h雙側(cè)MWT均明顯提高(P<0.05)，2 h時(shí)達(dá)高峰(P<0.05)，6 h時(shí)與給藥前無(wú)明顯差異(P>0.05)。見(jiàn)Fig 1。

Fig 1　The bilateral MWT of rats in five groups at each time point

compared with group Ⅰ, the bilateral MWT in group Ⅱ～Ⅴ was significantly decreased.vsthe MWT 1 h before intrathecal administration, the values at 1,2,4,6 h after administration of group Ⅱ and Ⅲ showed no marked difference. The ipsilateral MWT in group Ⅳ showed no significant difference at 1,2,4,6 h after administration, while the contralateral MWT was significantly increased at 1,2,4 h. In group Ⅴ the bilateral MWT was significantly improved at 1,2,4 h after administration.*P<0.05vsgroup Ⅰ;#P<0.05vsMWT 1h before intrathecal administration.

2.2GFAP測(cè)定結(jié)果

與Ⅰ組相比，Ⅱ和Ⅲ組雙側(cè)脊髓背角GFAP的染色明顯增強(qiáng)，Ⅳ組損傷側(cè)脊髓背角GFAP的染色明顯增強(qiáng)；與Ⅱ組相比，Ⅳ組對(duì)側(cè)脊髓背角GFAP的染色明顯減弱，損傷側(cè)無(wú)明顯變化，Ⅴ組雙側(cè)脊髓背角GFAP的染色均明顯減弱。GFAP陽(yáng)性細(xì)胞數(shù)的比較，與Ⅰ組相比，Ⅱ和Ⅲ組雙側(cè)脊髓背角GFAP的陽(yáng)性細(xì)胞數(shù)均明顯增多(P<0.05)；且Ⅱ、Ⅲ及Ⅳ組損傷側(cè)增加最明顯；與Ⅱ組相比，Ⅲ組及Ⅳ組損傷側(cè)脊髓背角GFAP陽(yáng)性細(xì)胞數(shù)無(wú)明顯變化(P>0.05)，而Ⅳ組對(duì)側(cè)脊髓背角GFAP的陽(yáng)性細(xì)胞數(shù)明顯減少(P<0.05)，Ⅴ組雙側(cè)脊髓背角GFAP陽(yáng)性細(xì)胞數(shù)均明顯降低(P<0.05)。見(jiàn)Fig 2，3。

Fig 2　GFAP expression in bilateral dorsal horns of the spinal cord in rats

compared with group Ⅰ(A), the bilateral expressions of GFAP in group Ⅱ(B) and Ⅲ(C) were significantly enhanced. In group Ⅳ(D), the ipsilateral expressions of GFAP were significantly enhanced; compared with group Ⅱ (B), the bilateral expressions of GFAP in group Ⅴ(E) were significantly decreased. In group Ⅳ (D), the contralateral expression was abviously reduced.

2.3脊髓背角TNF-α和IL-1β的表達(dá)

與Ⅰ組相比，Ⅱ～Ⅴ組雙側(cè)脊髓背角TNF-α和IL-1β的水平均明顯提高(P<0.05)；與Ⅱ組相比，Ⅲ和Ⅳ組損傷側(cè)脊髓背角TNF-α和IL-1β的水平無(wú)明顯變化(P>0.05)，對(duì)側(cè)脊髓背角TNF-α和IL-1β的水平明顯降低(P<0.05)，且Ⅳ組降低的更明顯；Ⅴ組雙側(cè)脊髓背角TNF-α和IL-1β的水平均明顯降低(P<0.05)。見(jiàn)Tab 1。

Fig 3　Number of GFAP-positive cells in bilateral spinal dosal horns of spinal cord in five±s)

compared with group Ⅰ，the number of GFAP-positive cells in the bilateral dosal horns of the spinal cord was significantly increased in group Ⅱ～Ⅴ; compared with group Ⅱ, the contralateral spinal dorsal horn of GFAP positive cells was significantly reduced in group Ⅳ, the bilateral spinal cord dorsal horn of GFAP-positive cells was significantly decreased in group Ⅴ.*P<0.05vsgroup Ⅰ;#P<0.05vsgroup Ⅱ.

3結(jié)論

在臨床和動(dòng)物模型中都發(fā)現(xiàn)有MIP的現(xiàn)象，臨床上如復(fù)雜區(qū)域疼痛綜合癥(complex regional pain syndrome, CRPS)、類(lèi)風(fēng)濕關(guān)節(jié)炎、纖維肌痛和NP；動(dòng)物模型如神經(jīng)損傷性疼痛、炎癥性疼痛及癌性疼痛[7-8]。MIP的典型特點(diǎn)是鏡像側(cè)的機(jī)械痛覺(jué)高敏，所以本研究主要觀察SNT大鼠的MWT值，研究發(fā)現(xiàn)，大鼠單側(cè)神經(jīng)損傷后，損傷側(cè)術(shù)后1 d即出現(xiàn)明顯的MWT值升高，對(duì)側(cè)術(shù)后5 d表現(xiàn)出明顯的MWT值升高。目前對(duì)MIP的機(jī)制仍然不清楚，被廣泛接受的有三大主要假說(shuō)：體液學(xué)說(shuō)、神經(jīng)學(xué)說(shuō)及膠質(zhì)細(xì)胞學(xué)說(shuō)，近年來(lái)，中樞神經(jīng)系統(tǒng)膠質(zhì)細(xì)胞的激活，通過(guò)GJ、鈣波及促炎性因子的釋放3個(gè)途徑在MIP的發(fā)生機(jī)制中受到越來(lái)越多的關(guān)注。GJ是相鄰細(xì)胞膜之間的連接通道，允許離子和小分子如cAMP、IP3、ATP和小分子肽類(lèi)在細(xì)胞間自由通過(guò)[9-11]。 在中樞神經(jīng)系統(tǒng)內(nèi)，GJ廣泛分布于星形膠質(zhì)細(xì)胞之間，星形膠質(zhì)細(xì)胞通過(guò)GJ廣泛偶聯(lián)形成一種星形膠質(zhì)細(xì)胞網(wǎng)絡(luò)。有學(xué)者發(fā)現(xiàn)星形膠質(zhì)細(xì)胞可以對(duì)因各種刺激而激活的神經(jīng)元細(xì)胞做出快速電反應(yīng)，表現(xiàn)為沿GJ傳播的Ca2+波[12]，傷害性刺激信號(hào)可能通過(guò)鈣波的形式在星形膠質(zhì)細(xì)胞GJ網(wǎng)絡(luò)中傳遞，引起疼痛的擴(kuò)散和傳播，進(jìn)而導(dǎo)致遠(yuǎn)處的神經(jīng)膠質(zhì)細(xì)胞和神經(jīng)元的活化，導(dǎo)致域外和鏡像效應(yīng)的產(chǎn)生。本實(shí)驗(yàn)發(fā)現(xiàn)，Ⅱ組大鼠術(shù)后10 d脊髓背角雙側(cè)GFAP的表達(dá)明顯增加，而鞘內(nèi)注射大劑量CBX后，脊髓背角雙側(cè)GFAP的表達(dá)明顯下降，提示GJ可能參與GFAP的激活。除此之外，促炎性因子在不同的NP模型及誘發(fā)或促進(jìn)NP中的重要作用，在許多實(shí)驗(yàn)研究中都已經(jīng)被證實(shí)[13-14]，之前我們的研究也發(fā)現(xiàn)，SNT大鼠術(shù)后損傷側(cè)脊髓背角TNF-α、IL-1β均明顯增高，鞘內(nèi)注射5 μg的CBX后MWT值明顯提高，同時(shí)TNF-α、IL-1β的表達(dá)也降低[15]。然而，大多數(shù)的研究都主要集中在神經(jīng)損傷側(cè)的神經(jīng)炎癥反應(yīng)，神經(jīng)損傷對(duì)側(cè)的炎癥反應(yīng)的研究卻比較少，所以本實(shí)驗(yàn)就觀察了大鼠雙側(cè)的神經(jīng)炎癥反應(yīng)情況，結(jié)果發(fā)現(xiàn)，SNT大鼠術(shù)后10 d雙側(cè)MWT明顯降低，同時(shí)雙側(cè)脊髓背角TNF-α和IL-1β的表達(dá)也明顯升高，鞘內(nèi)給予低劑量、中劑量及大劑量的CBX后，對(duì)側(cè)脊髓背角TNF-α和IL-1β的表達(dá)都降低，且隨著劑量的增加，炎性因子降低的越明顯，因此我們推測(cè)神經(jīng)損傷引起損傷側(cè)星形膠質(zhì)細(xì)胞的活化及增生，傷害性刺激信號(hào)通過(guò)GJ以鈣波的形式傳遞到對(duì)側(cè)，激活對(duì)側(cè)的星形膠質(zhì)細(xì)胞，引起對(duì)側(cè)促炎性細(xì)胞因子的釋放，這些細(xì)胞因子可以作為第二信使，進(jìn)一步作用于激活的神經(jīng)元和膠質(zhì)細(xì)胞，導(dǎo)致持續(xù)的痛覺(jué)過(guò)敏和異常性疼痛。

Tab 1　Expression of TNF-α and IL-1β in bilateral dorsal horns of spinal cord in five±s,pg·mg-1)

vsgroup Ⅰ, the expression of TNF-α and IL-1β at 2 h after administration in group Ⅱ,Ⅲ,Ⅳ and Ⅴ was significantly increased;vsgroup Ⅱ, the expression of TNF-α and IL-1β on the contralateral side in group Ⅲ and Ⅳ was significantly reduced, while the ipsilateral side in groupⅣ had no significant changes. The bilateral expressions of GFAP, TNF-α and IL-1β in group Ⅴ were significantly decreased.*P<0.05vsgroup Ⅰ;#P<0.05vsgroup Ⅱ.

本實(shí)驗(yàn)發(fā)現(xiàn)鞘內(nèi)注射CBX可逆性地緩解大鼠雙側(cè)MWT，結(jié)合星形膠質(zhì)細(xì)胞細(xì)胞和促炎性細(xì)胞因子在NP中的作用，因此，我們推測(cè)鞘內(nèi)注射CBX后抑制了細(xì)胞之間的GJ，從而阻斷傷害性信號(hào)向?qū)?cè)脊髓背角的傳遞，抑制對(duì)側(cè)膠質(zhì)細(xì)胞的激活及TNF-α和IL-1β等炎性細(xì)胞因子的釋放來(lái)逆轉(zhuǎn)痛覺(jué)高敏。

(致謝：本實(shí)驗(yàn)在南京軍區(qū)南京總醫(yī)院比較醫(yī)學(xué)科麻醉科實(shí)驗(yàn)室完成，在此衷心感謝我的導(dǎo)師李偉彥教授的耐心培養(yǎng)，實(shí)驗(yàn)室劉清珍老師的細(xì)心指導(dǎo)，許倩等各位同學(xué)的合作和幫助！)

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Effect of intrathecal injection of carbenoxolone on hyperalgesia in rats with L5 spinal nerve transaction

LI Xue-fei1,XU Qian1, WANG Fen1, ZHENG Man1, LIU Qing-zhen2,LI Wei-yan2

(1.DeptofAnesthesiology,AffiliatedHospitalofNanjingUniversityofTraditionalChineseMedicine,Nanjing210029,China;2.DeptofAnesthesiology,NanjingGeneralHospitalofNanjingMilitaryCommand,Nanjing210002,China)

Abstract:AimTo investigate the antagonistic effect of intrathecal injection of carbenoxolone(CBX) on neuropathic pain and its underlying mechanism.MethodsSixty male Sprague-Dawley rats were randomly divided into five groups(n=12): group I received sham surgery then treated with saline; group Ⅱ received SNT then treated with saline; group Ⅲ received SNT then treated with 0.05 μg CBX; group Ⅳ received SNT then treated with 0.5 μg CBX; group Ⅴ received SNT then treated with 5 μg CBX. Treatment was undertaken with 10 μl volume as a single intrathecal injection on postoperative day 10. Mechanical withdrawl thresholds were measured 1d before operation, 1, 3, 5, 7 and 10 d after surgery, 1 h before intrathecal administration, and 1, 2, 4, 6 h after intrathecal administration. Lumbar spinal cord was obtained 2 h after intrathecal administration to determine the expressions of GFAP by immunohistology and TNF-α,IL-1β by ELISA in bilateral spinal dorsal horns. ResultsCompared with the sham group, the bilateral MWT in group Ⅱ～Ⅴ was significantly decreased. Compared with the MWT 1 h before intrathecal administration on day 10, the values at 1, 2, 4, 6 h after administration of group Ⅱ and Ⅲ had no marked difference. The ipsilateral MWT in group Ⅳ had no significant difference at 1, 2, 4 h after administration, the contralateral MWT was significantly increased, whereas GFAP and TNF-α,IL-1β was significantly decreased in the spinal cord . In group Ⅴ the bilateral MWT was significantly improved at 1, 2, 4 h after administration, whereas GFAP and TNF-α,IL-1β were significantly decreased in the spinal cord.ConclusionsIntrathecal CBX can inhibit the development of bilateral MWT. The analgesic effect of CBX is implemented partly via suppressing the actation of GFAP and the realease of TNF-α,IL-1β in the spinal doral horn.

Key words:carbenoxolone；neuropathic pain；hyperalgesia；spinal cord dorsal horn；astrocyte；TNF-α；IL-1β

收稿日期:2016-02-01,修回日期:2016-03-25

基金項(xiàng)目：國(guó)家自然科學(xué)基金資助項(xiàng)目(No 30901399)

作者簡(jiǎn)介：李雪飛(1984-)，女，碩士，住院醫(yī)師，研究方向：星形膠質(zhì)細(xì)胞與神經(jīng)病理性疼痛，E-mail:xuefei9704@126.com;

doi:10.3969/j.issn.1001-1978.2016.06.024

文獻(xiàn)標(biāo)志碼：A

文章編號(hào)：1001-1978(2016)06-0863-05

中國(guó)圖書(shū)分類(lèi)號(hào)：R-332；R322.85；R392.12；R441.1；R741.041

網(wǎng)絡(luò)出版時(shí)間：2016-5-25 15:39網(wǎng)絡(luò)出版地址：http://www.cnki.net/kcms/detail/34.1086.R.20160525.1539.048.html

李偉彥(1965-)，男，博士，碩士生導(dǎo)師，研究方向：神經(jīng)病理性疼痛，E-mail:weiyanlee@yahoo.com.cn