Herpes simplex virus vector-mediated gene delivery of glutamic acid decarboxylase reduces detrusor overactivity in spinal cord-injured rats.
Herpes simplex virus vector-mediated gene delivery of glutamic acid decarboxylase reduces detrusor overactivity in spinal cord-injured rats.
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作者：
我们检测了编码67 kD谷氨酸脱羧酶(GAD67)基因产物的复制缺陷单纯疱疹病毒(HSV)载体是否能抑制脊髓损伤(SCI)大鼠的逼尿肌过度活动(DO)。脊髓化1周后，将表达GAD和绿色荧光蛋白(GFP)的HSV载体(HSV-GAD)注入膀胱壁。以未注射HSV的脊髓损伤大鼠(HSV未治疗)和注射LacZ报告基因HSV载体的SCI大鼠(HSV-LacZ)为对照。注射病毒3周后，三组均在清醒状态下进行连续膀胱测压。HSV-GAD组非排尿收缩次数和幅度明显低于HSV未治疗组和HSV-LacZ组(分别为40%~45%和38%~40%)，排尿效率高于HSV未治疗组和HSV-LacZ组，但三组间排尿压力差异无统计学意义。鞘内应用荷包牡丹碱可部分逆转HSV-GAD组NVCs数目和波幅的减少以及排尿效率的降低。与HSV-LacZ组相比，HSV-GAD组L6-S1背根节(DRG)的GAD67基因和蛋白表达显著增加，57%的DRG细胞呈GFP阳性，这些神经元表现出比HSV-LacZ组更强的GAD67免疫反应。这些结果表明，GAD基因治疗有效地抑制了脊髓损伤后的DO，主要是通过激活脊髓GABAA受体。因此，以单纯疱疹病毒为基础的GAD基因转移到膀胱传入通路可能是治疗神经源性DO的一种新途径。
We examined whether replication-defective herpes simplex virus (HSV) vectors encoding the 67 Kd form of the glutamic acid decarboxylase (GAD67) gene product, the gamma-aminobutyric acid (GABA) synthesis enzyme, can suppress detrusor overactivity (DO) in spinal cord injury (SCI) rats. One week after spinalization, HSV vectors expressing GAD and green fluorescent protein (GFP) (HSV-GAD) were injected into the bladder wall. SCI rats without HSV injection (HSV-untreated) and those injected with lacZ-encoding reporter gene HSV vectors (HSV-LacZ) were used as controls. Three weeks after viral injection, continuous cystometry was performed under awake conditions in all three groups. In the HSV-GAD group, the number and amplitude of non-voiding contractions (NVCs) were significantly decreased (40–45% and 38–40%, respectively) along with an increase in voiding efficiency, compared with HSV-untreated and HSV-LacZ groups, but micturition pressure was not different among the three groups. Intrathecal application of bicuculline partly reversed the decreased number and amplitude of NVCs, and decreased voiding efficiency in the HSV-GAD group. In the HSV-GAD group, GAD67 mRNA and protein levels were significantly increased in L6-S1 dorsal root ganglia (DRG) compared with the HSV-LacZ group while 57% of DRG cells were GFP-positive, and these neurons showed increased GAD67-like immunoreactivity compared with the HSV-LacZ group. These results indicate that GAD gene therapy effectively suppresses DO following SCI predominantly via activation of spinal GABAA receptors. Thus, HSV-based GAD gene transfer to bladder afferent pathways may represent a novel approach for the treatment of neurogenic DO.
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影响因子：
6.6
作者：
Seki, S;Sasaki, K;Yoshimura, N
通讯作者：
Yoshimura, N
DOI：
10.1073/pnas.93.21.11319
发表时间：
1996-10-15
影响因子：
11.1
作者：
Marconi, P;Krisky, D;Glorioso, JC
通讯作者：
Glorioso, JC
影响因子：
2.5
作者：
HOLSTEGE, G;GRIFFITHS, D;DALM, E
通讯作者：
DALM, E
影响因子：
11.2
作者：
Hao, SG;Mata, M;Fink, DJ
通讯作者：
Fink, DJ
影响因子：
12.4
作者：
Liu, J;Wolfe, D;Fink, DJ
通讯作者：
Fink, DJ