国际麻醉学与复苏杂志   2019, Issue (4): 0-0
    
丹参酮 ⅡA对长春新碱诱导大鼠机械痛敏和热痛敏形成的影响及其机制
付宝军, 姜静静, 黄玉琼, 李恒1()
1.广东省清远市人民医院
Effects of tanshinone ⅡA on the development of mechanical allodynia and heat hyperalgesia induced by vincristine in rats and its mechanism
 全文:
摘要:

目的 探讨丹参酮ⅡA对长春新碱诱导机械痛敏和热痛敏形成的影响及其机制。 方法 第一部分,采用随机数字表法将50只SD大鼠分为5组(每组10只):对照组(Control组)、化疗痛组[化疗诱导的神经病理性疼痛(chemotherapy-induced neuropathic pain, CINP)组]、化疗痛+丹参酮ⅡA 10 mg/kg组(CINP+Tan10组)、化疗痛+丹参酮ⅡA 20 mg/kg组(CINP+Tan20组)、化疗痛+丹参酮ⅡA 50 mg/kg组(CINP+Tan50组)。第二部分,采用随机数字表法将50只SD大鼠分为5组(每组10只):对照组(Control组)、化疗痛组(CINP组)、化疗痛+米诺四环素组(CINP+M组)、化疗痛+丹参酮ⅡA 20 mg/kg组(CINP+Tan20组)、化疗痛+丹参酮ⅡA 20 mg/kg+米诺四环素组(CINP+Tan20+M组)。隔日腹腔注射125 μg/kg长春新碱(共计4次)建立CINP动物模型,同时第1天开始连续7 d采用丹参酮ⅡA或米诺四环素治疗大鼠化疗痛。Control组和CINP组用生理盐水(腹腔注射,5 ml/kg)。第1次腹腔注射长春新碱前(T0)和腹腔注射长春新碱后1 d(T1)、腹腔注射长春新碱后3 d(T3)、腹腔注射长春新碱后5 d(T5)、腹腔注射长春新碱后7 d(T7)分别用机械刺激缩足反射阈值(paw mechanical withdrawal threshold, PMWT)和热刺激缩足反射潜伏期(paw withdraw thermal latency, PWTL)评价大鼠机械痛敏和热痛敏; T7时采用Western blot法检测脊髓胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)和白细胞分化抗原11b(cluster differentiation antigen 11b, CD11b)蛋白抗原(OX42)的表达;ELISA法检测脊髓TNF-α、IL-6含量变化。 结果 第一部分:与Control组比较,CINP组大鼠在T3、T5、T7 时PMWT和PWTL均明显降低,T7时大鼠脊髓OX42蛋白和TNF-α、IL-6相对含量明显升高(P<0.05);与CINP组比较,CINP+Tan20组、CINP+Tan50组在T3、T5、T7 时PMWT和PWTL均明显升高(P<0.05)。第二部分:与CINP组比较,CINP+M组、CINP+Tan20组、CINP+Tan20+M组大鼠在T3、T5、T7 时PMWT和PWTL均明显升高,T7时大鼠脊髓OX42蛋白和TNF-α、IL-6相对含量均明显降低(P<0.05)。 结论 丹参酮ⅡA能够延缓长春新碱诱导大鼠机械痛觉和热痛觉过敏形成,其作用机制可能与其抑制大鼠脊髓小胶质细胞活化,进而减少炎性细胞因子TNF-α、IL-6的含量有关。

关键词: 丹参酮ⅡA; 长春新碱; 痛觉过敏; 小胶质细胞
Abstract:

Objective To discuss the effects of tanshinone ⅡA on the mechanical allodynia and heat hyperalgesia induced by vincristine in rats and its mechanism. Methods Part 1, A total of 50 Sprague-Dawley(SD) rats were randomly divided into five groups (n=10): a control group (Control group), a CINP group (CINP group), a CINP+tanshinone ⅡA 10 mg/kg group (CINP+Tan10 group), a CINP+tanshinone ⅡA 20 mg/kg group (CINP+Tan20 group), a CINP+tanshinone ⅡA 50 mg/kg group (CINP+Tan50 group). Part 2, A total of 50 SD rats were randomly divided into five groups (n=10): a control group (Control group), a CINP group (CINP group), a CINP+minocycline group (CINP+M group), a CINP+tanshinone ⅡA 20 mg/kg group (CINP+Tan20 group), and a CINP+tanshinone ⅡA 20 mg/kg+minocycline group (CINP+Tan20+M group). Then, 125 μg/kg vincristine was intraperitoneally administered every two days (four times in total) to establish a rat model of chemotherapy-induced neuropathic pain (CINP). Meanwhile, the model rats were treated with tanshinone ⅡA or mino-tetracycline respectively for 7 d from Day 1 after exposure to vincristine. Rats in the control and CINP groups were given normal saline. Mechanical allodynia and heat hyperalgesia were evaluated by paw mechanical withdrawal threshold (PMWT) and paw thermal withdrawal latency (PWTL) respectively before first exposure to vincristine (T0) and on Days 1 (T1), 3 (T3), 5 (T5) and 7 (T7) after exposure to vincristine. At T7, the expression of glial fibrillary acidic protein (GFAP) and cluster differentiation antigen 11b (CD11b) antigen OX42 were detected by Western Blot. The content of tumor necrosis factor (TNF-α) and interleukin 6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA). Results Part 1: Compared with the control group, the CINP group presented remarkable decreases in PMWT and PWTL at T3, T5, and T7, and marked increases in the expression of OX42, TNF-α and IL-6 at T7(P<0.05). Compared with the CINP group, there were marked increases in PMWT and PWTL in the CINP+Tan20 and CINP+Tan50 groups at T3, T5 and T7 (P<0.05). Part 2: Compared with the CINP group, there were marked increases in PMWT and PWTL at T3, T5 and T7 in the CINP+M, CINP+Tan20 and CINP+Tan20+M groups, while obvious decreases were observed in the expression of OX42, TNF-α and IL-6 in the CINP+M, CINP+Tan20 and CINP+Tan20+M groups at T7(P<0.05). Conclusions Tanshinone ⅡA can delay the development of vincristine-induced mechanical allodynia and heat hyperalgesia, which may be related to the inhibition of spinal microglia activation, so as to reduce the content of TNF-α and IL-6.

Key words: Tanshinone ⅡA; Vincristine; Hyperalgesia; Microglia