摘要
目的 研究正己烷周围神经毒效应与红细胞膜收缩蛋白（又叫血影蛋
白，spectrin，Sp）共价交联的关系。
方法 SD 大鼠正己烷灌胃，分 0、168、675、2700 mg/kg四组，5d/
周、连续 8 周，观察动物体重、脏器指数(心、肝、脾、肺、肾、脑和睾
丸)、生化指标(谷草转氨酶、谷丙转氨酶、总蛋白、白蛋白、球蛋白、总
胆红素、直接胆红素、碱性磷酸酶、谷氨酰转肽酶、尿素氮、肌酐、尿酸、
葡萄糖、乳酸脱氢酶、磷酸肌酸激酶、磷酸肌酸激酶同工酶、羟丁酸脱氢
酶、总胆汁酸、胆碱酯酶)、混合神经传导速度（mixednervesconduction
velocity，MNCV）和斜板法测肌力功能，光、电镜观察胫神经。双缩脲
法测抽提的红细胞膜蛋白浓度，聚丙烯酰胺凝胶电泳(sodiumdodecyl
sulfate-polyacrylamide gel electrophoresis，SDS-PAGE)后银染，光密度扫
描计算蛋白含量，用 Western blot 鉴定 Sp。
结果 染毒 8 周后，2700mg/kg组动物后肢瘫痪、瘫痪率为 80％，
体重减轻、MNCV 减慢、肌力下降（P<0.05），主要生化指标未见明显变
化，肾、心、肝、肺、脑指数明显上升（P<0.05），胫神经光镜下表现为
轴索数目稀疏、髓鞘变薄，电镜下轴索大小不一、部分肿胀、神经丝密度
增加、部分髓鞘回缩。红细胞膜蛋白浓度为：5.88±0.89g/L，电泳后银染
见 240kD条带之上有异常条带，经 Western blot 证实为 Sp 共价交联产物，
带形为连续“拖尾”状。共价交联部分占总 Sp 的百分比呈时间-剂量-效应
关系。
结论 正己烷 2700mg/kg连续灌胃 8 周，可复制出致周围神经毒性
动物模型，出现后肢瘫痪和周围神经轴索病变。并可引起 Sp 共价交联，
其作为正己烷周围神经毒性效应替代标志物具有可行性，但仍需深入研
究。
Objective: To study the relationship between covalent cross-linking
erythrocyte spectrin and n-hexane neurotoxicity. Method: SD rats were given
n-hexane of 0、168、675、2700 mg/kg for 8 weeks 5d/w）ig. General condition,
（
body weight，biochemical indexes (AST, ALT, TP, ALB, T-BIL, D-BIL, ALP,
GGT, BUN, CRE, UA, GLB, GLU, LDH, CK, CK-MB, HBDH, TBA, CHE)
and the main organic indexes (heart, liver, kidney, brain, spleen, lung and
spermary) were observed or tested. Mixed nerves conduction velocity
（MNCV）and muscle tension tested by slide board were carried out. The
muscle branch of the posterior tibial nerve was observed under light
microscope and electron microscope. Protein was eluted from erythrocytes
and then the concentration was tested. It was isolated by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), dyed by silver
staining , quantified by scanner and identified by Western blot. Results: In the
group of 2700mg/kg, at the end of the test ( 8 weeks after the test started),
animals had intoxication symptoms such as hindlimb paralysis, decreasing of
body weight, muscle tension and MNCV. Biochemical indexes did not
changed significantly. Organic indexes（kidney, heart, liver, lung, brain）
increased significantly (P<0.05). In the tibial nerve, morphological changes
such as axons decreased in numbers, degenerated were observed under light
microscope. In the axon, diversiform, swollen, demyelination and malaligned
neruofilaments were showed under electron microscope. The concentration
erythrocytes protein of was 5.88±0.89g/L. A high-molecular-weight protein
band for about 410 kD was showed on SDS-PAGE. It was identified by
Western blot as the covalent cross-linking spectrin (a,?-heterodimer) with a
long “tailling”. There was time-dose-effect relationship with the increase of
ＩＩ
covalent cross-linking spectrin. Conclusion: Animal model of neurotoxicity
induced by n-hexane could be successfully duplicated with hindlimb paralysis,
neuropathy and erythrocyte spectrin covalent cross-linking. It was feasible for
covalent cross-linking erythrocyte spectrin to be the surrogate biomarker of
neurotoxicity induced by n-hexane. But it was necessary to further explore it.
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