【摘要】：目的觀察注射右美托咪定(Dexmedetomidine,DEX)預(yù)處理對在體大鼠脊髓缺血再灌注損傷(Spinal cord ischemia reperfusion injury,SCIRI)后神經(jīng)元自噬和下肢運動功能的影響。方法 SD大鼠隨機(jī)分為4組(每組30只):假手術(shù)組(Sham組)、缺血再灌注損傷組(IR組,鞘內(nèi)注射30μL生理鹽水)、Dex組(缺血前3 d鞘內(nèi)注射右美托咪定10μg/30μL)、Dex+ATIP組(缺血前3 d鞘內(nèi)注射10μg右美托咪定+10μg阿替美唑共30μL)。Sham組僅暴露主動脈弓而不結(jié)扎,其他各組開胸后無創(chuàng)動脈夾夾閉主動脈弓14 min后再開放,建立SCIRI模型。各組手術(shù)前均于L5-6鞘內(nèi)置管并連續(xù)注射3 d。損傷后48 h內(nèi)每12小時取5只大鼠,采用Tarlov法評價大鼠下肢運動功能;HE染色觀察和計數(shù)損傷后48 h脊髓前角神經(jīng)元形態(tài)和數(shù)量;Western blot和RT-PCR法測定損傷后48 h大鼠脊髓組織中自噬相關(guān)蛋白Beclin1、LC3蛋白及mRNA含量。結(jié)果與Sham組比較,術(shù)后各觀察點IR組下肢運動功能明顯受損,Tarlov評分下降,脊髓組織中Beclin1、LC3蛋白及mRNA的表達(dá)均明顯降低(P0.05);損傷前3 d鞘內(nèi)注射10μg DEX(Dex組)可改善缺血再灌注損傷后下肢運動功能,提高Tarlov評分,改善脊髓Beclin1、LC3 mRNA蛋白的表達(dá)(P0.05),而Dex+ATIP組與IR組比較差異無統(tǒng)計學(xué)意義(P0.05)。損傷后48 h,HE染色顯示,Sham組脊髓前角神經(jīng)元輪廓清晰,結(jié)構(gòu)完整,無出血、水腫等異常表現(xiàn);IR組、Dex+ATIP組細(xì)胞質(zhì)中的尼氏體消失,神經(jīng)元胞核出現(xiàn)固縮或溶解,廣泛空泡形成;而Dex組神經(jīng)元形態(tài)明顯規(guī)整,且正常神經(jīng)元數(shù)量增多。結(jié)論鞘內(nèi)注射右美托咪定預(yù)處理通過上調(diào)Beclin1、LC3蛋白和基因表達(dá),增加神經(jīng)元自噬,改善大鼠缺血再灌注后下肢運動功能。
[Abstract]:Aim to observe the effects of pretreatment with right metametidine (Dexmedetomidine,DEX) on neuronal autophagy and lower limb motor function after spinal cord ischemia-reperfusion injury (Spinal cord ischemia reperfusion injury,SCIRI) in rats in vivo. Methods SD rats were randomly divided into 4 groups (30 rats in each group): sham operation group (Sham group), ischemia reperfusion injury group (IR group, intrathecal injection of 30 渭 L saline), Dex (intrathecal injection of right metomide 10 渭 g / 30 渭 L) 3 days before ischemia). In Dex ATIP group (intrathecal injection of 10 渭 g right metametrimidine 10 渭 g atimeimazole) 3 days before ischemia, only aortic arch was exposed without ligation in 30 渭 L). Sham group, and non-invasive aortic arch was closed for 14 min after thoracotomy in other groups. The SCIRI model was established by intrathecal injection of 10 渭 g right metometrimidine 10 渭 g atimezole. All the patients in each group were placed in the sheath of L _ 5 ~ 6 before operation and injected continuously for 3 d. Within 48 hours after injury, 5 rats were taken out every 12 hours to evaluate the motor function of the lower extremities by Tarlov method, and the morphology and number of neurons in the anterior horn of spinal cord were observed and counted by HE staining at 48 hours after injury. The contents of Beclin1,LC3 protein and mRNA in spinal cord tissue of 48 h after injury were determined by Western blot and RT-PCR. Results compared with Sham group, the motor function of lower extremities and the expression of Beclin1,LC3 protein and mRNA in spinal cord were significantly decreased in IR group after operation (P0.05). The scores of Tarlov were decreased, and the expression of Beclin1,LC3 protein and mRNA in spinal cord were significantly decreased (P0.05). Intrathecal injection of 10 渭 g DEX (Dex 3 days before injury could improve lower limb motor function, increase Tarlov score and improve the expression of Beclin1,LC3 mRNA protein in spinal cord after ischemia-reperfusion injury (P0.05). There was no significant difference between Dex ATIP group and IR group (P0.05). 48 hours after injury, HE staining showed that neurons in the anterior horn of the spinal cord in the Sham group were clear in outline, complete in structure, without hemorrhage and edema. In IR group, the Nissl body disappeared in the cytoplasm of, Dex ATIP group, the nucleus of neurons became pyknotic or dissolved, and extensive vacuoles were formed, while the morphology of neurons in Dex group was obviously regular, and the number of normal neurons was increased. Conclusion Intrathecal injection of right metomidine can improve the motor function of lower limbs after ischemia-reperfusion by up-regulating the expression of Beclin1,LC3 protein and gene and increasing the autophagy of neurons.
【作者單位】： 中國醫(yī)科大學(xué)附屬第一醫(yī)院麻醉科;
【基金】：遼寧省科學(xué)技術(shù)計劃項目(2012408002) 國家自然科學(xué)基金(81271370)
【分類號】：R614

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