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目的:探讨多次应用不同剂量的一氧化氮(NO)对福尔马林炎性痛中脊髓背角神经元Bcl-2、Bax表达的影响。方法:连续4 d给大鼠各进行鞘内注射不同剂量的一氧化氮前体左旋精氨酸(L-arginine,L-Arg)10μg/d(低L-Arg组)、250μg/d(高L-Arg组)或一氧化氮合酶(nitric oxide synthase,NOS)抑制剂Nω-硝基-L-精氨酸甲酯(Nω-nitro-L-arginine methylester,L-NAME)2700μg/d(L-NAME组),鞘内注射生理盐水(NS组)作为对照,各给药1次/d。随后于大鼠右后肢脚掌皮下注射福尔马林(2%,100μl),4 h后分别做免疫组化和Western Blot检测脊髓背角Bcl-2和Bax蛋白的表达。结果:免疫组化结果显示,Bcl-2和Bax表达分布于两侧脊髓背角,但主要位于脊髓背角浅层,各组在注射福尔马林同侧均明显高于对侧。Western Blot结果显示Bcl-2和Bax蛋白表达含量的比值,在各实验组与对照组比较,低L-Arg组Bcl-2/Bax比值明显升高,而高L-Arg组和L-NAME组Bcl-2/Bax比值均明显降低。bcl-2为抑制细胞凋亡的基因,而bax为促进细胞凋亡的基因。结论:在炎性痛模型中,低剂量的NO以促进抑凋亡基因的表达为主,而高剂量的NO以及NO生成不足均以促进促凋亡基因的表达为主,进而影响炎性痛的发生。
Objective: To investigate the effects of multiple doses of nitric oxide (NO) on the expression of Bcl-2 and Bax in spinal dorsal horn neurons in formalin-induced inflammatory pain. Methods: The rats were intrathecally injected with L-arginine (L-Arg) 10μg / d (low L-Arg), 250μg / d L-Arg group) or Nω-nitro-L-arginine methylester (L-NAME) 2700 μg / d L-NAME group), and intrathecal NS (NS group) as control. Each dose was once / d. Subsequently, formalin (2%, 100μl) was injected subcutaneously into the soles of the right hindlimbs of rats. After 4 hours, the expression of Bcl-2 and Bax protein in spinal dorsal horn was detected by immunohistochemistry and Western Blot respectively. Results: The results of immunohistochemistry showed that the expression of Bcl-2 and Bax were located in the dorsal horn on both sides of the spinal cord, but mainly located in the superficial layer of dorsal horn of spinal cord. Each group was significantly higher on the same side of formalin injection than the contralateral side. The results of Western Blot showed that the ratio of Bcl-2 / Bax protein expression in each group was significantly higher than that in the control group, while the ratio of Bcl-2 / Bax in low L-Arg group was significantly higher than that in the control group Bcl-2 / Bax ratio were significantly lower. bcl-2 is a gene that inhibits apoptosis, and bax is a gene that promotes apoptosis. CONCLUSIONS: In the inflammatory pain model, NO at a low dose promotes the expression of anti-apoptotic genes, whereas high NO and NO production are mainly contributed to promote the expression of pro-apoptotic genes and further affect inflammatory pain happened.