左归丸对睡眠剥夺大鼠认知功能的影响

Effect of Zuogui pill on cognitive function in sleep-deprived rats

目的评价左归丸对睡眠剥夺大鼠认知功能的影响。方法SPF级健康雄性SD大鼠30只,8周龄,体重200 g左右,采用随机数字表法分成5组(n=6):对照组(C组)、睡眠剥夺组(SD组)、左归丸低剂量组(L组)、左归丸中剂量组(M组)及左归丸高剂量组(H组)。采用多平台水环境法构建大鼠睡眠剥夺模型。L组、M组和H组左归丸剂量分别为2.42、4.84、9.68 g/kg,每天上午10点灌胃给药,灌胃液体量每次4 ml,C组和SD组大鼠采用纯水灌胃,L组、M组和H组按照左归丸相应剂量,配制成药液灌胃,1次/d,持续7 d。采用Morris水迷宫实验评估大鼠认知功能,第2次行为学实验结束后,采集眼球血样,随后断头处死大鼠取海马,采用ELISA法检测血清IL-1β、IL-6和TNF-α浓度及海马组织MDA、谷胱甘肽过氧化物酶(GSH-Px)和SOD含量;采用Western blot法检测海马组织IL-1β、IL-6和TNF-α的表达。结果与C组比较,其余各组游泳总路程和逃避潜伏期延长,原象限停留时间缩短,穿越原平台位置次数减少,血清和海马组织IL-1β、IL-6、TNF-α水平升高,MDA含量升高,GSH-Px和SOD含量降低(P<0.05);与SD组比较,L组、M组和H组大鼠游泳总路程和逃避潜伏期缩短,原象限停留时间延长,穿越原平台位置次数增加,血清和海马组织IL-1β、IL-6、TNF-α水平降低,MDA含量降低,GSH-Px和SOD含量升高(P<0.05)。结论左归丸可改善睡眠剥夺大鼠的认知功能,其机制与抑制炎性反应和提高抗氧化能力有关。

Objective To evaluate the effect of Zuogui pill on cognitive function in sleep-deprived rats.Methods Thirty SPF Sprague-Dawley male rats,aged 8 weeks,weighing about 200 g,were divided into 5 groups(n=6 each)using a random number table method:control group(group C),sleep deprivation group(group SD),low-dose Zuogui pill group(group L),medium-dose Zuogui pill group(group M)and high-dose Zuogui pill group(group H).The model of sleep deprivation was developed by the multi-platform water environment method.In L,M and H groups,the doses of Zuogui pill were 2.42,4.84 and 9.68 g/kg,respectively,Zuogui pill was administered by intragastric gavage at 10:00 am every day,and the volume of fluid for intragastric gavage was 4 ml each time.In C and SD groups,pure water was used for intragastric gavage.In L,M and H groups,the liquid was prepared according to the corresponding doses of Zuogui pill for intragastric gavage,once a day for 7 consecutive days.The cognitive function was evaluated by the Morris water maze test.After the end of the second behavioral testing,blood samples from the eyeballs were collected,and then the rats were sacrificed by decapitation,and brains were removed and the hippocampus was obtained for determination of the concentrations of serum interleukin-1beta(IL-1β),IL-6 and tumor necrosis factor-alpha(TNF-α)and contents of malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)in hippocampal tissues(by enzyme-linked immunosorbent assay)and the expression of IL-1β,IL-6 and TNF-αin hippocampal tissues(by Western blot).Results Compared with group C,the total swimming distance and escape latency were significantly prolonged,the time of staying at the original quadrant and the number of crossing the platform were decreased,the levels of serum IL-1β,IL-6 and TNF-αin serum and hippocampal tissues were increased,the content of MDA was increased,and the contents of GSH-Px and SOD were decreased in the other groups(P<0.05).Compared with group SD,the total swimming distance and escape latency were significantly shortened,and the time of staying at the original quadrant was prolonged,and the number of crossing the platform was increased,the levels of serum IL-1β,IL-6 and TNF-αin serum and hippocampal tissues were decreased,the content of MDA was decreased,and the contents of GSH-Px and SOD were increased in L,M and H groups(P<0.05).Conclusions Zuogui pill can improve the cognitive function in sleep-deprived in rats,and the mechanism is related to inhibition of inflammatory responses and improvement in the antioxidant capacity.