[Effect of herbal cake-separated moxibustion on behavioral stress reactions and blood lactic acid level and muscular AMPK/PGC-1α signaling in rats with chronic fatigue syndrome].

To observe the effect of herbal cake-separated moxibustion (HCSM) on serum lactic acid (BLA) level and AMPK/PGC-1α signaling pathway in the quadriceps femoris in chronic fatigue syndrome (CFS) rats, so as to explore its mechanisms underlying improvement of CFS.According to the random number table, 50 SD rats were divided into blank control, model, HCSM, sham HCSM and medication (herbal medicine gavage) groups, with 10 rats in each group. The CFS model was established by using chronic restraint and exhaustive swimming, alternately, once daily for 21 days. The herbal cake was made of Xiaoyao Powder (Mental Ease Powder, composed of [Danggui (Radix Angelicae Sinensis), Baishao (Radix Paeoniae Alba), Chaihu (Radix Bupleuri), Fuling (Poria), Baizhu (Rhizoma Atractylodis, Macrocephalae), etc.]. The HCSM was applied to "Shenque" (CV8), "Guanyuan "(CV4), bilateral "Zusanli" (ST36) and "Qimen" (LR14), 5 moxa-cones for each acupoint, once daily for 10 days. For sham HCSM, the excipient was instead of herbal cake, and the same 5 moxa-cones was given as the HCSM group. Rats of the medication group received gavage of Xiaoyao Powder suspension (60 mg·kg-1), once daily for 10 days. The open field test and tail suspension test were conducted for determining the animals' locomotor activity. The blood sample was taken from the abdominal aorta under anesthesia for assaying the levels of serum BLA, chemokine ligand CXCL9 and β-endorphin (EP) by ELISA. Bilateral quadriceps femoris were sampled for observing histopathological changes after staining with conventional H.E. technique, and for detecting the expression levels of phosphorylated AMP-activated protein kinase (p-AMPK) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) by using immunohistochemistry.Compared with the blank control group, the number of rearing and horizontal grid-crossing times, struggling times of tail suspension test were significantly decreased (P<0.05), and the immobility time was obviously prolonged (P<0.05) in the model group. Compared with the model group, both HCSM and medication groups had a significant increase of rearing, horizontal grid-crossing times and struggling times (P<0.05), and the immobility time had a significant decrease (P<0.05). But there were no significant differences in the total movement distance among the 5 groups (P>0.05), and in the 5 indexes of behavioral measurements between the HCSM and medication groups (P>0.05). The sham HCSM could also evidently increase the struggling times and reduce the immobility time (P<0.05). The contents of serum BLA, CXCL9 and β-EP were obviously higher in the model group than in the blank control group (P<0.05), as well as remarkably lower in the HCSM and medication groups than in the model group (P<0.05). Whereas the expression levels of muscular p-AMPK and PGC-1α were considerably lower in the model group than in the blank control group (P<0.05), and significantly increased in both HCSM and medication groups relevant to the model group (P<0.05). Compared with the sham HCSM group, the contents of BLA, CXCL9 and β-EP in serum of the HCSM group and contents of CXCL9, β-EP in medication group were significantly decreased (P<0.05), and the protein expressions of p-AMPK and PGC-1α in quadriceps femoris in both HCSM and medication groups were significantly increased (P<0.05). H.E. staining showed smaller intercellular space, uneven cytoplasmic staining in some muscle fibers, nucleus pyknosis and condensation, and inflammatory cell infiltration in the model group, which was milder in both HCSM and medication groups.HCSM can mitigate the stress behavioral state in CFS rats, which may be related with its functions in lowering the levels of serum BLA, CXCL9 and β-EP, and activating AMPK/PGC-1α signaling pathway (balancing energy metabolism) in the quadriceps femoris.目的:观察隔药饼灸对慢性疲劳综合征(CFS)大鼠血乳酸(BLA)及腺苷酸活化蛋白激酶/过氧化物酶体增殖物激活受体-γ共激活因子-1α(AMPK/PGC-1α)信号通路的影响, 探讨隔药饼灸治疗CFS的部分作用机制。方法:SD大鼠随机分为空白组、模型组、隔药饼灸组、假药饼灸组和中药灌胃组, 每组10只。采用慢性束缚与力竭游泳交替进行21 d的复合应激造模方法复制CFS大鼠模型。隔药饼灸组以逍遥散组方的药物制成药饼, 于大鼠“神阙”“关元”“足三里”“期门”进行隔药饼灸, 每穴灸5壮;假药饼灸组用赋形剂制备的假药饼, 每穴灸5壮;中药灌胃组选择逍遥散悬浊液(60 mg·kg-1)灌服。以上干预均每日1次, 共10 d。旷场实验与悬尾实验评价各组大鼠行为学变化;ELISA法检测大鼠血清中BLA、趋化因子CXC配体9(CXCL9)及β-内啡肽(β-EP)含量;HE染色法观察大鼠股四头肌病理形态;免疫组织化学法检测大鼠股四头肌磷酸化(p)-AMPK与PGC-1α蛋白表达水平。结果:与空白组比较, 模型组大鼠旷场实验站立次数与水平跨格数均显著减少(P<0.05);与模型组、假药饼灸组比较, 隔药饼灸组与中药灌胃组大鼠站立次数与水平跨格数显著增加(P<0.05)。与空白组比较, 模型组大鼠悬尾实验挣扎次数显著减少(P<0.05), 不动时间明显延长(P<0.05);与模型组比较, 隔药饼灸组、假药饼灸组与中药灌胃组大鼠挣扎次数明显增加(P<0.05), 不动时间显著缩短(P<0.05)。与空白组比较, 模型组大鼠血清中BLA、CXCL9及β-EP含量显著升高(P<0.05), 股四头肌部分纤维结构破坏, 胞核固缩、凝聚, 股四头肌p-AMPK与PGC-1α蛋白表达显著降低(P<0.05);与模型组比较, 隔药饼灸组和中药灌胃组血清BLA、CXCL9及β-EP含量显著降低(P<0.05), 肌纤维排列恢复, 细胞间隙有所增大, 股四头肌p-AMPK与PGC-1α蛋白表达显著升高(P<0.05), 假药饼灸组肌纤维排列仍稀松杂乱;与假药饼灸组比较, 隔药饼灸组血清BLA、CXCL9及β-EP含量显著降低(P<0.05), 股四头肌p-AMPK与PGC-1α蛋白表达显著升高(P<0.05), 中药灌胃组血清CXCL9、β-EP含量降低(P<0.05), 股四头肌p-AMPK、PGC-1α蛋白表达升高(P<0.05)。结论:对CFS大鼠进行隔药饼灸治疗可调控血清BLA、CXCL9与β-EP含量, 同时激活AMPK/PGC-1α信号通路, 从而改善机体氧化应激状态, 维持能量代谢的相对平衡。.