灵芝孢子和一氧化氮合酶抑制剂对大鼠脊髓损伤后背核、红核神经元存活及轴突再生的影响

EFFECTS OF GANODERMA SPORE AND L-NNA ON THE NEURONAL SURVIVAL AND AXONAL REGENERATION IN CLARKE'S NUCLEUS AND RED NUCLEUS AFTER RAT SPINAL CORD INJURY

目的探讨灵芝孢子和一氧化氮合酶(NOS)抑制剂(L-NNA)联合应用能否促进脊髓半横断后受损伤的背核和红核神经元存活及其轴突再生。方法30只成年雌性大鼠被分为对照组、L-NNA组、灵芝孢子低剂量组、灵芝孢子低剂量+L-NNA组、灵芝孢子高剂量组和灵芝孢子高剂量+L-NNA组。灵芝孢子高剂量+L-NNA组动物在T11脊髓段半横断后注射L-NNA和胃饲高剂量灵芝孢子。结果脊髓半横断后30 d,对照组L1脊髓损伤侧背核神经元数量减少,NOS表达阳性。L-NNA组和灵芝孢子低剂量组损伤侧背核神经元增加,但NOS表达降低。灵芝孢子低剂量+L-NNA组和灵芝孢子高剂量组损伤侧背核神经元明显增加,NOS表达显著降低。灵芝孢子高剂量+L-NNA组损伤侧背核神经元最多,NOS表达最低,而且有些神经元胞体被荧光金标记。对照组和L-NNA组损伤侧红核神经元密度降低。灵芝孢子低剂量组和灵芝孢子低剂量+L-NNA组损伤侧红核神经元密度提高。灵芝孢子高剂量组和灵芝孢子高剂量+L-NNA组损伤侧红核神经元密度明显提高。结论灵芝孢子和L-NNA都能促进脊髓半横断后受损伤背核神经元的存活,两者联合应用能更好地促进受损伤的背核神经元存活及其轴突再生。灵芝孢子也能促进受损伤的红核神经元存活。

Objective Whether combination of ganoderma spore and L-NNA（NOS inhibitor） could promote the neuronal survival and axonal regeneration in Clarke＇s nucleus （CN） and red nucleus （RN） following spinal cord hemisection was explored. Methods Thirty aduh female rats were divided into control group, L-NNA group, low dosage of ganoderma spore group, low dosage of ganoderma spore ＋ L-NNA group, high dosage of ganoderma spore group and high dosage of ganoderma spore ＋ L-NNA group. L-NNA was intraperitoneally injected and high dosage of ganoderma spore was irrigated into the rat＇s stomach after bemisection of T11 spinal cord segment in high dosage of ganoderma spore ＋ L-NNA group. Results Thirty days after spinal cord hemisection, the number of CN neurons was decreased and some neurons were expressing NOS on lesioned side of L1 spinal cord in control group. The number of CN neurons on lesioned side of L1 spinal cord was increased in L-NNA group and low dosage of ganoderma spore group, but the number of NOS-positive neurons was decreased. In low dosage of ganoderma spore ＋ L-NNA group and high dosage of ganoderma spore group, the number of CN neurons was significantly increased and NOS-positive neurons were also significantly decreased on lesioned side of L1 spinal cord. In high dosage of ganoderma spore ＋ L-NNA group, there were most CN neurons surviving and least NOS-positive CN neurons, and some neurons were labeled by flourogold on lesioned side of LI spinal cord. The density of RN neurons on lesioned side was decreased in control group and L-NNA group. The density of RN neurons on lesioned side was increased in low dosage of ganoderma spore group and low dosage of ganoderma spore ＋ L-NNA group. The density of RN neurons on lesioned side was significantly increased in high dosage of ganoderma spore group and high dosage of ganoderma spore ＋ L-NNA group. Conclusion Ganoderma spore or L-NNA can promote the survival of injuried CN neurons after spinal cord hemisection. Combination of ganoderma spore and L-NNA can better promote injuried CN neuronal survival and axonal regeneration. Ganoderma spore can also enhance injuried RN neuronal survival.