神经纤维纵切及同时显示ATP-ase和ChE活性的组织化学方法

在介绍制作新鲜冰冻神经纤维切片简单、有效方法的同时,报告了在切片上同时显示三磷酸腺苷酶(ATP-ase)及胆碱酯酶(ChE)活性的组织化学方法,这对观察神经纤维内两种酶的相互关系和生理或病理性的细胞化学改变是有意义的。     

氯丙烯染毒引起大鼠脊髓神经细胞胆碱酯酶活力改变的组织化学研究

研究了正常和氯丙烯染毒(剂量为100mg/kg)后大鼠脊髓神经细胞胆碱酯酶(CHE)的活力变化。正常对照组脊髓前角和中央管两侧神经细胞胞浆内CHE染成靛蓝色;染毒后,CHE活力下降,即在少数神经细胞内显现少量的淡靛蓝色颗粒。CHE活力降低以腰髓最重。毛细血管周边CHE活力与对照组相似。说明氯丙烯致神经细胞CHE活力下降是由于细胞体受影响,并非直接抑制了CHE的活力。对细胞体的影响可能是因其直接作用于周围神经纤维,引起微管和神经微丝变性,阻断了轴浆运输,使轴突破坏或阻断的信息传到细胞体或从神经终末端到细胞体的信息反馈中断,影响了CHE的合成。     

Effect of Bupivacaine application on cholinesterase activities,total protein- and albumin concentration in serum and cerebrospinal fluid

The rationale of this study was to determine whether Bupivacaine used for spinal anesthesia alters the specific secretory activity of nerve cells and/or the function of the blood/cerebrospinal fluid barrier. Four groups were assessed: (1) patients undergoing spinal anesthesia using Bupivacaine for lower limb surgery, (2) spinal Bupivacaine anesthesia without subsequent surgery, (3) local facet joint infiltration using Bupivacaine, and (4) general anesthesia for lower limb surgery without Bupivacaine application. Cholinesterase activities, total protein- and albumin concentrations in serum as well as in cerebrospinal fluid were significantly decreased after surgical intervention under spinal Bupivacaine anesthesia but remained unchanged following spinal Bupivacaine application without surgery. No significant correlation was found between Bupivacaine dosage and parameter alteration. There was no influence of intrathecal Bupivacaine application on the albumin ratio cerebrospinal fluid/serum, nor was there any significant alteration of total protein- or albumin concentrations and butyrylcholinesterase activity in the serum as a result of local injection of Bupivacaine to facet joints. These serum parameters were reduced after surgery under general anesthesia. Alterations of serum- and cerebrospinal fluid parameters investigated after surgery are not related to Bupivacaine application but to effects linked to operative treatment, i.e. suppressed secretory cell activity or protein depletion owing to blood loss. We conclude that the secretory function of cholinesterase-releasing nerve cells is not affected by spinal application of Bupivacaine. The blood/cerebrospinal fluid barrier remains intact.     

γ-谷氨酰转肽酶/胆碱酯酶比值预测慢性乙型肝炎患者肝纤维化程度的效能评价*

目的 评价γ-谷氨酰转肽酶/胆碱酯酶比值(GCR)与γ-谷氨酰转肽酶/血小板比值(GPR)预测慢性乙型肝炎(CHB)患者肝纤维化程度的效能。方法 本研究纳入1335例HBeAg阳性和1026例HBeAg阴性CHB患者。采用Scheuer评分系统评估肝组织纤维化程度,将肝纤维化分期≥S2和＝S4分别定义为显著纤维化和肝硬化。结果 在HBeAg阳性患者中,发现886例显著肝纤维化,286例肝硬化,在HBeAg阴性患者中,发现556例显著肝纤维化,202例肝硬化;在HBeAg阳性患者,GCR预测显著纤维化的ROC曲线下面积(AUC)为0.770 (95%CI:0.747～0.793),显著大于GPR预测【0.757 (0.733～0.780), P<0.05】,GCR预测肝硬化的AUC 为0.816 (95%CI:0.794～0.837),接近GPR预测【0.808 (0.786～0.829)】;在HBeAg阴性患者,GCR预测显著纤维化的AUC为0.761 (95%CI:0.733～0.787),接近GPR预测【0.758 (0.731～0.784)】,GCR预测肝硬化的AUC为0.838 (95%CI:0.814～0.860),接近GPR预测 【0.829 (0.804～0.851)】;以GCR>0.100和GPR>0.500为截断点,预测HBeAg阳性患者显著纤维化的特异度为81.5%和80.6%,预测HBeAg阴性患者显著纤维化的特异度为81.5%和79.4%,预测HBeAg阳性患者肝硬化的灵敏度为84.3%和81.5%,预测HBeAg阴性患者肝硬化的灵敏度为79.2%和82.7%。结论 无论在HBeAg阳性和阴性CHB患者,GCR预测显著纤维化和肝硬化的效能高于或接近GPR。     

大鼠杀虫双染毒后全血胆碱酯酶活力的变化

目的 探讨沙蚕毒系农药杀虫双对胆碱酯酶（ChE）活力是否有抑制及抑制的程度。方法 Wistar大鼠分为4组，分别以杀虫双或甲胺磷按1／16、1／8、1／4、1／2LD50剂量经口染毒，于染毒前及染毒后1／2、1、2、4、24h取大鼠尾尖血，以改良Ellman法测定ChE活力。结果 杀虫双1／16LD50剂量组染毒大鼠血ChE活力未受影响；随染毒剂量的加大，ChE活力逐步下降，1／2LD50剂量组大鼠血ChE活力抑制率为正常组的35.9%，差异有显著性（P＜0.01)。甲胺磷1／16LD60染毒剂量组大鼠血ChE活力抑制率为正常对照组的42.4%；随染毒剂量的加大，大鼠血ChE的活力进一步下降，1／2LD50剂量组大鼠血ChE活力抑制率达正常对照组的52.9%。各剂量杀虫双染毒组的ChE活力均高于对应的甲胺磷染毒组，差异有显著性（P＜0.01)。结论 大鼠经口杀虫双染毒后，在较高染毒剂量下可以抑制ChE活力，但抑制程度明显弱于甲胺磷。     

Alterations of Na+/K+-ATPase,cholinergic and antioxidant enzymes activity by protocatechuic acid in cadmium-induced neurotoxicity and oxidative stress in Wistar rats

BackgroundThis study assessed the possible protective mechanisms of protocatechuic acid (PCA) against cadmium (Cd)-induced oxidative stress and neurotoxicity in rats.MethodsMale wistar strain rats weighing between 150–160 g were purchased and acclimatized for two weeks. The rats were divided into seven groups of seven each; NC group received normal saline, CAD group received 6 mg/kg of Cd-solution, CAD + PSG group received Cd-solution and prostigmine (5 mg/kg), CAD + PCA-10 and CAD + PCA-20 groups received Cd-solution and PCA (10 mg/kg and 20 mg/kg) respectively, PCA-10 and PCA-20 groups received 10 mg/kg and 20 mg/kg PCA each. Animals were administered normal saline, Cd and PCA daily by oral gavage for 21 days. After which the animals were sacrificed, the brain excised, homogenized and centrifuged. The activities of enzymes (Na⁺/K⁺-ATPase, cholinesterases, catalase, glutathione peroxidase, superoxide dismutase) and levels of oxidative stress markers (lipid peroxidation and reduced glutathione) linked to neurodegeneration were subsequently assessed.ResultsSignificant (p < 0.05) alterations in the enzyme activities and levels of oxidative stress markers were observed in CAD group when compared to the NC group. However, the activities of the enzymes were reversed in CAD + PSG and CAD + PCA groups.ConclusionsPCA may protect against cadmium-induced neurotoxicity by altering the activities of Na⁺/K⁺-ATPase, acetylcholinesterase, butyrylcholinesterase and endogenous antioxidant enzymes.     

氯丙烯对神经纤维ATPase和ChE活力影响的组织化学观察

以ATPase和ChE联合染色的组织化学方法观察氯丙烯对神经纤维的ATPase和ChE活力的影响。结果表明,ATPase活力略有降低,ChE活力显著降低或消失。结合前文用电子显微镜观察轴突内微管和神经微丝变性和数量减少的结果,可以证明氯丙烯损伤微管和神经微丝,阻断轴浆运输,引起了周围神经的病变。     

Experimental alcoholic neuropathy in the rat: Histological and electrophysiological study on the myoneural junctions and the peripheral nerves

Summary Peripheral nerves and myoneural junctions of the tibialis anterior muscle of the rat were studied histologically and electrophysiologically after various periods of peroral ethanol treatment. Histochemical distributions of non-specific cholinesterase (ns. ChE; E.C. 3.1.1.8) and acetylcholinesterase (AChE; E.C. 3.1.1.7) activity of the muscle were normal during the first 3 months of daily ethanol drinking. After 5 months of exposure to 10–25% (v/v) ethanol as the sole drinking fluid, pathological ns. ChE activity was seen sporadically along the intramuscular nerves with slight ultrastructural changes in the Schwann cells. After 7 months of ethanol treatment there was further increased pathological ns. ChE activity in the intramuscular nerves while the AChE activity remained normal in the muscle. More prominent ultrastructural changes were seen in the Schwann cells namely swelling and vacuolization of the cytoplasm and dilatation of the rough endoplasmic reticulum. Increased numbers of small axons were also seen. After 9.5 months on alcohol marked increase in the ns. ChE activity was observed along most of the intramuscular nerves. AChE activity of the myoneural junctions was only sporadically weakened.A slight slow-down in the conduction velocity of the large myelinated size A fibers was observed in the animals on alcohol from 7–9.5 months, whereas the conduction velocity of the smaller myelinated B fibers was not appreciably changed. The present experiment indicates that progressive neuropathy can be induced in rats by oral alcohol feeding along with the normal laboratory diet. The first pathological changes were seen in the Schwann cells and could well be followed by the methods employed. The present experimental model can possibly be used in future studies concerning the development of toxic polyneuropathy.Supported by the Finnish Foundation for Alcohol Studies     

Activity of acetylcholinesterase and unspecific cholinesterase during differentiation of somites in mouse embryos

During the development of somites in mouse embryos, widespread activity of unspecific cholinesterase (BuChE) was demonstrated after prolonged incubation. Independent of their position, all somite cells and their derivatives (dermatome, myotome and sclerotome) exhibited enzyme activity in the perinuclear space and in the endoplasmic reticulum. The plasmalemma did not show any enzyme activity. Differentiation of the sclerotome into vertebrae was accompanied by a reduction of BuChE. However, a low enzyme reaction was still present in the first typical differentiated chondroblasts. Notochordal cells were detectable by their high BuChE content. This was also found in cells already showed severe degeneration. In addition to BuChE, acetylcholinesterase (AChE) was first visible on day 9 of embryonic development in newly formed myotubes of the myotomes. Some hypotheses on the functional significance of embryonic BuChE are discussed in the light of these results.     

氯丙烯染毒引起运动终板的胆碱酯酶活力和肌肉组织变化的组织化学研究

氯丙烯染毒后经过一定时间出现神经性肌萎缩。肌萎缩与运动终板胆碱酯酶活力降低大体上成比例。染毒23日后出现同型肌纤维聚集成簇及簇内运动终板增多。表明肌萎缩的局限区内肌纤维进行神经再支配,再支配的神经来自健康运动神经元的轴突分支,可能产生新的运动终板。