谷氨酸通过调控脊髓背角GABA_(B)R2参与慢性胰腺炎痛的机制研究  

Glutamate Participates in Chronic Pancreatitis Pain by Regulating GABA_(B)R2 Expression in Spinal Dorsal Horn

作　　者：徐卉红 邱海波 王莉琴 朱成成 陆智杰 XU Hui-hong;QIU Hai-bo;WANG Li-qin;ZHU Cheng-cheng;LU Zhi-jie(Department of Anesthesiology,the Third Affiliated Hospital of Naval Medical University,Shanghai 200438,China;Department of Anesthesiology,No.905 Hospital of PLA NAVY,Shanghai 200052,China)

机构地区：[1]海军军医大学第三附属医院麻醉科,上海200438 [2]中国人民解放军海军第九0五医院麻醉科,上海200052

出　　处：《南昌大学学报（医学版）》2022年第4期10-15,共6页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(31371084,82171232)。

摘　　要：目的探讨谷氨酸通过调控脊髓背角GABA_(B)R2参与慢性胰腺炎(chronic pancreatitis,CP)疼痛的机制研究,为缓解慢性胰腺炎痛寻找新的理论依据。方法将42只Wistar雄性大鼠(约200 g)分为对照组(n=12)和CP组(n=30)。CP组尾静脉单次注射二丁基二氯化合物(DBTC,8 mg·kg^(-1))构建CP模型,对照组注射溶剂。通过HE染色明确胰腺组织病理特征改变。利用Von Frey电子测痛仪测定每组大鼠在注射前和注射后3、7、14、21 d的腹部机械痛。Western-blot及免疫荧光组织化学检测大鼠T8—T12节段脊髓背角、体外培养脊髓背角神经元上GABA_(B)R2的表达。结果CP模型大鼠胰腺组织可见散在腺泡细胞肿胀坏死,广泛的炎症细胞浸润及大量纤维结缔组织增生。与对照组比较:CP组大鼠腹部机械痛阈值明显下降,并持续至DBTC注射后21 d(P<0.05,P<0.001);在DBTC注射后7、14、21 d脊髓背角GABA_(B)R2的表达明显减少(P<0.001)。体外实验结果表明,原代培养的脊髓背角神经元经20、40μmol·L^(-1)谷氨酸处理后,GABA_(B)R2的表达均明显减少(P<0.001)。结论谷氨酸通过调控脊髓背角神经元GABA_(B)R2的表达是CP疼痛发生的机制之一。Objective To explore the mechanism of glutamate participating in chronic pancreatitis(CP)pain by regulating GABA_(B)R2 expression in the spinal dorsal horn,and to provide new theoretical basis for relieving CP pain.Methods Forty-two male Wistar rats weighing about 200 g were divided into control group(n=12)and CP group(n=30).The CP model was established by a single tail vein injection with 8 mg·kg^(-1)dibutyltin dichloride(DBTC).The control group only received the solvent.The pathological changes in pancreatic tissue were confirmed by HE staining.Abdominal mechanical allodynia was measured using Von Frey electronic pain meter before and 3,7,14,21 days after injection.The expression of GABA_(B)R2 in the T8-T12 segments of spinal dorsal horn and cultured spinal dorsal horn neurons was detected by Western blot and immunofluorescence histochemistry.Results HE staining showed scattered acinar cell swelling and necrosis,extensive inflammatory cell infiltration,and massive fibrous connective tissue hyperplasia in the pancreatic tissue of CP rats.Compared with the control group,the abdominal mechanical pain threshold obviously decreased and lasted to the 21st day after DBTC injection in CP group(P<0.05,P<0.001).Furthermore,the expression of GABA_(B)R2 in the spinal dorsal horn was also down-regulated at 7,14 and 21 days after DBTC injection group(P<0.001).In vitro experiment showed that GABA_(B)R2 expression was significantly reduced in primarily cultured spinal dorsal horn neurons after treatment with 20 and 40μmol·L^(-1)glutamate(P<0.001).Conclusion Glutamate is involved in the pathogenesis of CP pain by regulating the expression of GABA_(B)R2 in spinal dorsal horn neurons.

关 键 词：慢性胰腺炎 疼痛 脊髓背角 Γ-氨基丁酸受体 谷氨酸 动物 实验 大鼠 

分 类 号：R-332[医药卫生]