Zou Wang-Yuan, Yang Yong, Guo Qu-Lian
Department of Anesthesiology, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2008 Jun;33(6):481-7.
To investigate the antinociceptive effect of periaqueductal gray (PAG) administration of herpes simplex virus type-1(HSV-I) amplicon vector-mediated human preproenkephalin gene (HPPE).
Sprague-Dawley rats weighting 260 to approximately 320 g were randomly divided into pHSVIRES-HPPE-LacZ (SHPZ) group, pHSVIRES-LacZ (SHZ) group, and saline (NS) group which included 3 d,1 week,2 week,3 week,4 week,5 week, and 6 week groups (n=51). The rats were anesthetized with intraperitoneal chloral hydrate (300 to approximately 350) mg/kg. Rats were PAG delivered with recombinant HSV-I amplicon vector SHPZ, SHZ or NS. One week after PAG administration 9 rats in each group were sacrificed and lumber segment of the spinal cord was removed for determination of expression of LacZ by X-gal staining and HPPE mRNA expression by reverse transcription-polymerase chain reaction and L-enkephalin content by radioimmunoassay in PAG. Formalin 50 microL (5%) was injected into the left hindpaw, and pain intensity scoring (PIS) was used to assess the antinociceptive effect.
After in vivo transferring, neurocyte demonstrated strong positive signals with X-gal immunohistochemical staining. The expression of HPPE mRNA was detected in PAG after administration of SHPZ. PAG delivery of SHPZ showed antinociceptive effect on formalin-induced pain for 6 weeks compared with SHZ group.
This amplicon virus can transfer HPPE into rat PAG neural cells and make it express efficiently. PAG administration of SHPZ can produce significant analgesic effect on formalin-induced pain in rats for 5 weeks.
研究导水管周围灰质（PAG）注射单纯疱疹病毒I型（HSV-I）扩增载体介导的人前脑啡肽原基因（HPPE）的抗伤害感受作用。
将体重260至约320 g的Sprague-Dawley大鼠随机分为pHSVIRES-HPPE-LacZ（SHPZ）组、pHSVIRES-LacZ（SHZ）组和生理盐水（NS）组，每组又分为3天、1周、2周、3周、4周、5周和6周亚组（n = 51）。大鼠腹腔注射水合氯醛（300至约350）mg/kg麻醉。将重组HSV-I扩增载体SHPZ、SHZ或NS注入大鼠PAG。PAG注射1周后，每组处死9只大鼠，取出脊髓腰段，通过X-gal染色测定LacZ的表达，通过逆转录-聚合酶链反应测定HPPE mRNA的表达，并通过放射免疫测定法测定PAG中亮脑啡肽的含量。向大鼠左后爪注射50 μL（5%）福尔马林，采用疼痛强度评分（PIS）评估抗伤害感受作用。
体内转染后，神经细胞经X-gal免疫组化染色显示强阳性信号。SHPZ注射后在PAG中检测到HPPE mRNA的表达。与SHZ组相比，PAG注射SHPZ对福尔马林诱导的疼痛具有6周的抗伤害感受作用。
该扩增病毒可将HPPE导入大鼠PAG神经细胞并使其高效表达。PAG注射SHPZ可对大鼠福尔马林诱导的疼痛产生5周的显著镇痛作用。
