普瑞巴林联合吴茱萸碱对神经病理性疼痛大鼠细胞因子和T细胞的影响

Effect of Pregabalin and Evodiamine on Cytokines and T Cells in a Rat Model of Neuropathic Pain

目的:观察普瑞巴林联合吴茱萸碱对神经病理性疼痛大鼠免疫细胞的影响。方法:SD大鼠30只。随机分为5组(每组6只):假手术组(S组)、对照组(C组)、普瑞巴林组(P组)、吴茱萸碱组(Q组)和普瑞巴林-吴茱萸碱复合物(F组)。大鼠暴露左侧L5脊神经并结扎,建立脊神经结扎(SNL)模型,S组大鼠暴露左侧L5脊神经,但不结扎。建立SNL模型7 d后P组腹腔注射普瑞巴林(5 mg/kg),Q组腹腔注射吴茱萸碱注射液(5 mg/kg),F组腹腔注射普瑞巴林-吴茱萸碱复合物(5 mg/kg),S和C组腹腔注射生理盐水(2 m L/kg),连续给药7 d,观察测量大鼠神经损伤侧的疼痛行为学的变化。给药7 d后取主动脉血3 m L检测大鼠肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、5-羟色胺(5-HT)、去甲肾上腺素(NE)和T淋巴细胞亚群中T细胞分化群4(CD4^+)、T细胞分化群8(CD8^+)的水平。结果:模型制成后S组大鼠活动如常,其它四组大鼠SNL 1周后,步态及姿势出现不同程度的异常;P组、Q组、F组三组动物SNL第10天开始,上述症状逐渐缓解,尤其F组症状表现轻微。给药7 d后C组TNF-α为(101.75±15.46)μg/L、IL-6为(32.98±6.64)μg/L、NE为(78.14±4.38)pg/mL、5-HT为(6.21±1.87)μmol/L,较S组TNF-α[(55.14±13.28)μg/L]、IL-6[(18.16±5.98)μg/L]、NE[(23.65±2.21)pg/mL]、5-HT[(1.89±0.76)μmol/L]显著升高(P<0.01);F组TNF-α为(68.54±17.65)μg/L、IL-6为(20.21±4.23)μg/L、NE为(33.08±3.85)pg/mL、5-HT为(2.64±1.38)μmol/L较C组明显降低(P<0.05)。给药7 d后C组CD4^+为(16.43±1.68)ng/mL、CD8^+为(11.26±2.31)ng/mL较S组CD4^+[(25.28±1.56)ng/mL]、CD8^+[(14.05±2.45)ng/mL]显著降低(P<0.01);F组CD4^+为(32.45±3.45)ng/mL、CD8^+(21.47±1.08)ng/mL较C组升高(P<0.01或P<0.05)。结论:普瑞巴林联合吴茱英碱对神经病理性疼痛大鼠具有较好的协同镇痛作用,优于各自单独使用;两者合用能够使神经病理性疼痛家兔炎性细胞因子的表达下调,并能延缓神经病理性疼痛大鼠T淋巴细胞亚群CD4^+、CD8^+的下降。

Objective To observe the effect of pregabalin and evodiamine on immune cells in a rat model of neuropathic pain. Methods Adult male Sprague-Dawley （SD） rats were subjected to left L5 spinal nerve ligation model （SNL）. In a sham operation, the left L5 spinal nerve was exposed but not ligated. Total 30 mice were randomly assigned to 5 groups （6 rats in each group）： sham operation （Group S）, control （Group C）, pregabalin （Group P）, evodiamine （Group Q） and pregabalin combined evodiamine （Group F）. Seven days after SNL, rats in different groups were treated as follows： intraperitoneal injection of pregabalin （5 mg/kg） （Group P）, intraperito- neal injection of evodiamine （5 plexes （Group F）, Rats in group mg/kg） （Group Q）, intraperitoneal injection of pregabalin and evodiamine com- C were received intraperitoneal injection of physiological saline （2 mL/kg）. All treatment lasted for 7 days. From the beginning of the experiment, the changes of pain behavior in rats were observed every day. In the end, blood （3ml） was sampled from the aorta and examined for serum tu- mor necrosis factor-alpha （TNF-alpha）, interleukin -6 （IL-6）, 5-serotonin （5-HT）, and norepinephrine （NE） and for CD4＋ and CD8＋ T lymphocytes. Results Spinal nerve ligation changed gait and posture in all related groups. However, gait and posture improved in Group P, Group Q, and Group F 10 days after SNL, especially in Group F. In Group F, serum levels of TNF-tx, IL-6, NE, and 5-HT were significantly lower than those in the Group C （P 〈 0.05）. In Group C, CD4＋ and CD8＋ T cells were significantly less than Group S. Group F had more CD4＋ and CD8＋ T cells than Group C （P 〈 0.01 or 〈 0.05）. Conclusion Joint use of pregabalin and evodiamine in rat model of neuropathie pain had good synergistic analgesic effect. The combination of the two medicines can decrease the expression of TNF-alpha, IL-6, 5-HT, NE and delay the decrease in CD4＋, CD8＋ T lymphocyte subsets in rats with pathological pain.