白藜芦醇通过沉默信息调节因子2相关酶1/过氧化物酶体增殖物激活受体γ共激活因子1α信号通路减缓紫杉醇诱发神经痛

Resveratrol alleviated paclitaxel-induced neuropathic pain by silent information regulator 2-related enzymes 1/peroxisome proliferator-activated receptor-gamma coactivator 1α signaling pathway

目的观察沉默信息调节因子2相关酶1/过氧化物酶体增殖物激活受体γ共激活因子1α(SIRT1/PGC1α)信号通路在白藜芦醇对紫杉醇诱发神经痛的影响。方法清洁级雄性SD大鼠(体重180~200 g),取鞘内置管成功大鼠50只,随机分为5组(n=10):正常对照组(C组),紫杉醇组(P组),白藜芦醇组(R组),白藜芦醇+紫杉醇组(R+P组),EX-527(SIRT1抑制剂)+白藜芦醇+紫杉醇组(E+R+P组)。第1、3、5、7天经腹腔注射紫杉醇2 mg/kg,第1~14天每天鞘内注射EX-527 10 μg/10 μl或腹腔注射白藜芦醇40 mg/kg。5组大鼠分别于紫杉醇给药前1 d(T0)、给药后8 d(T1)、给药后14 d(T2)测定50%机械缩足痛阈值(PWT)。紫杉醇给药后第15天取纹状体,透射电镜观察线粒体结构;原位缺口末端标记法(TUNEL)法测定纹状体细胞凋亡;Western blot法测定纹状体SIRT1和PGC1α蛋白表达;酶联免疫吸附试验(ELISA)法测定纹状体白细胞介素(IL)-1β和IL-10浓度。结果在T2时间点,与C组PWT(13.1±1.0)比较,P组和E+R+P组PWT分别降低至6.2±1.0和5.6±3.4(P<0.05);与P组比较,R+P组PWT升高至14.3±2.7(P<0.05)。P组和E+R+P组线粒体肿胀、空泡,而R+P组线粒体损伤减轻。与C组比较,P组纹状体细胞凋亡增加(P<0.05);与P组比较,R+P组细胞凋亡减少(P<0.05)。与C组的SIRT1(0.650±0.074)和PGC1α(0.590±0.057)蛋白表达比较,P组分别下调至0.320±0.032和0.200±0.067(P<0.05);与P组比较,R+P组分别上调至0.530±0.010和0.530±0.041(P<0.05)。与C组IL-1β[(7.57±0.09) pg/ml]、IL-10[(32.65±0.16) pg/ml]浓度比较,P组纹状体IL-1β浓度增加至(15.11±0.20) pg/ml(P<0.05],IL-10浓度降低至(12.92±0.17) pg/ml(P<0.05);与P组比较,R+P组IL-1β浓度降低至(9.42±0.13) pg/ml(P<0.05),IL-10浓度增高至(24.77±0.12) pg/ml(P<0.05)。结论白藜芦醇可通过激活SIRT1/PGC1α信号通路减轻紫杉醇诱发神经痛,这可能与其减少纹状体内细胞凋亡、抑制炎性反应、改善线粒体损伤有关。

Objective To investigate the effect of resveratrol (Res) on paclitaxel-induced neuropathic pain in rats by silent information regulator 2-related enzymes 1/peroxisome proliferator-activated receptor-gamma coactivator 1α(SIRT1/PGC1α) signaling pathway. Methods Fifty male Sprague-Dawley rats (weighing 180-200 g), after intrathecal intubation, were randomly assigned into five groups (n=10): control group (C group), paclitaxel group (P group), Res group (R group), Res pretreated group (R+ P group) and EX-527 (the specific inhibitor of SIRT1)+ Res pretreated group (E+ R+ P group). Paclitaxel (2 mg/kg) was intraperitoneally injected on the 1st, 3rd, 5th and 7 day. Res (intraperitoneal injection, 40 mg/kg) and EX-527 (intrathecal injection, 10 μg/10 μl) were administered daily from the 1st day to the 14th day. We measured 50% paw withdrawal mechanical threshold (PWT) at three time points: 1 day before chemotherapy (T0), 8 days after chemotherapy (T1) and 14 days after chemotherapy (T2). The corpus striatum was harvested at the 15th day after paclitaxel intraperitoneal injection. Transmission electron microscope was applied to observe the mitochondrial histomorphology in five groups. The cell apoptosis in the corpus striatum was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The expression of SIRT1 and PGC1α was detected using Western blotting. The levels of IL-1β and IL-10 in the corpus striatum were determined by enzyme linked immunosorbent assay (ELISA). Results At T2, as compared with C group (13.1±1.0), the PWT in P group and E+ R+ P group was decreased to (6.2±1.0) and (5.6±3.4) respectively (P<0.05);as compared with P group, the PWT in R+ P group was increased to (14.3±2.7)(P<0.05). In P group and E+ R+ P group, we observed swellings and vacuolations in the mitochondria, and these changes in R+ P group was alleviated. As compared with C group, the apoptosis cells in the corpus striatum were increased in P group (P<0.05), and they were significantly reduced in R+ P group as compared with P group (P<0.05). As compared with levels of SIRT1 (0.650±0.074) and PGC1α(0.590±0.057) in C group, the expression levels of SIRT1 and PGC1α in P group were decreased to (0.320±0.032) and (0.200±0.067) respectively (P<0.05);as compare with P group, the expression levels of SIRT1 and PGC1α in R+ P group were increased to (0.530±0.010) and (0.530±0.041) respectively (P<0.05). As compared with IL-1β(7.57±0.09) pg/ml and IL-10 (32.65±0.16) pg/ml in C group, the levels of IL-1β in P group were increased to (15.11±0.20) pg/ml (P<0.05), and those of IL-10 in P group were decreased to (12.92±0.17) pg/ml (P<0.05);as compared with P group, the level of IL-1β in R+ P group was significantly decreased to (9.42±0.13) pg/ml (P<0.05), and that of IL-10 in P group was significantly increased to (24.77±0.12) pg/ml (P<0.05). Conclusion Res alleviates paclitaxel-induced neuralgia, which is related with activation of SIRT1/PGC1α signaling pathway, reduction of cell apoptosis, inhibition of inflammation, and improvement of mitochondrial damage.