首页
The Protective Effect of Olanzapine on the Hippocampal Neuron of Depression Model Rats via Inhibiting NLRP3 Inflammasome Activation
-
摘要:目的 探究奥氮平(OLA)对抑郁症模型大鼠海马神经元的影响及作用机制。方法 将大鼠分为对照组、慢性不可预见性应激(CUS)组、OAL (0.5、1、2 mg/kg)组、si-Atg5及OAL (2 mg/kg)+si-Atg5组，旷场实验及糖水偏好实验评估大鼠行为学表现，Tunnel检测细胞凋亡，ELISA检测白介素(IL)-1β、IL-18质量浓度，Western blot检测cleaved Caspase-3，cleaved Caspase-9，自噬相关蛋白LC3、Beclin1、P62，炎症小体NLRP3及cleaved Caspase-1表达水平。结果 0.5、1、2 mg/kg OAL均可增加CUS大鼠自发活动总路程、糖水消耗量及偏好率，降低大鼠IL-18血清质量浓度，海马CA3区凋亡细胞百分比，cleaved Caspase-9、cleaved Caspase-1、NLRP3表达；0.5 mg/kg OAL对cleaved Caspase-3表达及IL-1β血清质量浓度无影响，1、2 mg/kg OAL可降低cleaved Caspase-3表达及IL-1β血清质量浓度。si-Atg5可减小CUS大鼠自发活动总路程、糖水消耗量及偏好率，提高cleaved Caspase-3、cleaved Caspase-9、cleaved Caspase-1、NLRP3表达，并减弱2 mg/kg OAL产生的影响。同时，0.5、1、2 mg/kg OAL均可提高大鼠海马CA3区LC3Ⅱ/LC3Ⅰ比值及Beclin1表达；0.5 mg/kg OAL对P62表达无影响，1、2 mg/kg OAL可降低P62表达。si-Atg5可降低LC3Ⅱ/LC3Ⅰ比值及Beclin1的表达, 并减弱2 mg/kg OAL产生的作用。结论 OAL可通过抑制NLRP3炎症小体激活对CUS大鼠海马神经元产生保护作用。Abstract:Objective To determine the impact olanzapine (OLA) on the hippocampal neuron of model rats with depression.Methods Rats were divided into five groups: control, chronic unpredicted stress (CUS), OAL (0.5, 1, 2 mg/kg), si-Atg5, and OAL (2 mg/kg)+si-Atg5. Open field and sucrose preference tests were performed to evaluate rat behaviors. Cell apoptosis was detected with Tunnel. The concentrations of interleukin (IL)-1β and IL-18 were determined by ELISA. The expressions of cleaved Caspase-3, cleaved Caspase-9, LC3, Beclin1, P62, NLRP3 and cleaved Caspase-1 were measured by Western blot.Results OAL (0.5, 1, 2 mg/kg) increased the total moving distance, sucrose consumption and preference rate of CUS rats, and decreased serum IL-18, cell apoptosis and the expressions of cleaved Caspase-9, cleaved Caspase-1 and NLRP3 in the CA3 region of hippocampus. Although OAL (1, 2 mg/kg) decreased the expression of cleaved Caspase-3 and serum IL-1β, OAL (0.5 mg/kg) showed no detectable effects. Si-Atg5 decreased the total moving distance, sucrose consumption and preference rate of CUS rats, enhanced the expressions of cleaved Caspase-3, cleaved Caspase-9, cleaved Caspase-1 and NLRP3, and weakened the effect of OAL (2 mg/kg). OAL (0.5, 1, 2 mg/kg) also increased the LC3Ⅱ/LC3Ⅰ ratio and the expression of Beclin1 in the CA3 region of hippocampus. OAL (1, 2 mg/kg) reduced the expression of p62, but not when it was reduced to 0.5 mg/kg. Si-Atg5 reduced the LC3Ⅱ/LC3Ⅰ ratio and the expression of Beclin1, and weakened the function of OAL (2 mg/kg).Conclusion OAL can protect the hippocampal neuron of CUS rats via inhibiting NLRP3 inflammasome activation.
-
Keywords:
- Olanzapine /
- Depression /
- NLRP3 inflammasome
-
抑郁症是常见的精神疾病，以情绪低落、快感缺失、长期失眠及认知障碍为主要临床表现，重度抑郁症患者往往具有自杀倾向[1]。随着都市生活节奏的加快，人们承受的压力日益增大，抑郁症发病率逐年增高。全球目前约有3.5亿人受到抑郁症的困扰，探索抑郁症的治疗方法成为医学界研究热点[2]。奥氮平(OAL)是一种作用于多巴胺和5-羟色胺受体的非典型抗精神病药，可通过拮抗多巴胺受体减少精神病患者的阳性症状，如幻觉、妄想、言语、思维及行为紊乱，通过拮抗5-羟色胺受体减少精神病患者的阴性症状，如情绪低落、冷漠、快感缺失及注意力不足[3]。同时，OAL也是治疗双相抑郁症的主要药物之一[4]，其与氟西汀联合使用可显著地缓解双相抑郁症状[5]。目前，OAL在国内临床应用时间并不长，其对患者海马神经元产生的影响还有待深入研究。因此，本实验通过建立大鼠慢性不可预见性应激(CUS)模型，对OAL在海马神经元中的影响及作用机制进行深入探究。
1. 材料与方法
1.1 实验动物和试剂
42只3周龄SPF级SD大鼠购自河南远大生物制药有限公司，许可证号：SYXK(豫)2017-0014；OAL购自美国Eli Lilly; Tunnel试剂盒及BCA试剂盒购自上海碧云天；白介素(IL)-1β及IL-18 ELISA试剂盒购自美国Invirtrogen；实验所用蛋白抗体均购自美国Cell Signaling Technology；二抗均购自北京中杉金桥；si-Atg5购自上海生工生物。
1.2 大鼠分组及处理
将42只SD大鼠分为对照，CUS、OAL (0.5、1、2 mg/kg)、si-Atg5及OAL (2 mg/kg)+si-Atg5组，每组6只。除对照组以外，其余各组均建立大鼠CUS抑郁症模型，OAL用药浓度[6]及大鼠CUS抑郁症模型建立方法[7]参考既往文献。通过旷场实验、糖水偏好实验检测造模情况。造模成功后，OAL(0.5、1、2 mg/kg)组分别灌胃给予0.5、1、2 mg/kg OAL，si-Atg5组用si-Atg5腺病毒转染大鼠，OAL (2 mg/kg)+si-Atg5组用si-Atg5腺病毒转染大鼠并灌胃给予2 mg/kg OAL，CUS组则给予等量生理盐水，每日1次，持续给予3周后，收集大鼠外周血，处死大鼠后剥离大脑海马CA3区组织，石蜡包埋脑组织，用于后期检测。
1.3 旷场实验
造模前后将各组大鼠单独置于透明观察室，适应10 min后，记录其5 min内自发活动路程。观察后，每只大鼠用清洁剂清洗并干燥。
1.4 糖水偏好实验
将各组大鼠单独置于饲养笼中，进行预实验，第一日，笼中放置2瓶1%蔗糖溶液，第二日用自来水取代其中1瓶蔗糖溶液。适应后，断粮水供应24 h，于上午9点进行正式实验。笼中分别放置100 mL自来水及蔗糖溶液各1瓶，1 h后，记录蔗糖溶液及自来水消耗体积，糖水偏好率=糖水消耗量/(水消耗量+糖水消耗量)×100%。
1.5 HE染色
将脑海马区组织石蜡包埋切片，然后将切片脱蜡、水洗、苏木精液染色、水洗；再用1%盐酸酒精分化，水洗；0.6%氨水返蓝，水洗；0.5%伊红液染色，水洗；接着脱水，二甲苯透明；最后中性树胶封片，每组取6张切片，在400倍显微镜观察海马区组织损伤情况。
1.6 Tunnel检测
将大鼠大脑海马组织石蜡切片脱蜡修复，内源性过氧化物酶作用5 min后，PBS洗涤。滴加Tunnel检测液37 ℃避光孵育1 h后，PBS洗涤。利用DAB显色液避光显色，苏木精复染。暗室内观察切片，显微镜视野下，棕色为凋亡细胞，蓝色为正常细胞。每组取6张切片于400倍镜下观察。凋亡率(%)=凋亡细胞数/细胞总数×100%。
1.7 ELISA检测大鼠血清IL-1β和IL-18质量浓度
利用ELISA试剂盒测定IL-1β、IL-18血清质量浓度，步骤参考试剂盒说明书。避光显色，利用酶标仪读取吸光度(A)值，根据标准样品曲线公式计算各样品A值对应的蛋白质量浓度。
1.8 Western blot检测海马CA3区脑组织蛋白水平
RIPA裂解海马CA3区脑组织，提取总蛋白。BCA试剂盒将蛋白定量并调平。取蛋白30 g，10% SDS-PAGE将蛋白分离，湿转法将蛋白转移至PVDF膜，脱脂奶粉室温封闭2 h，各蛋白对应一抗4 ℃孵育过夜，二抗37 ℃ 1 h，最后曝光显色，以GAPDH为内参，薄膜由Bio-rad凝胶DocEZ成像仪拍摄，利用Image J软件分析检测Capase-3、-9、NLRP3、LC3、Beclin1和P62蛋白条带的灰度值与内参蛋白带的灰度值比值。
1.9 统计学方法
所有数据先进行正态分布和方差齐性分析，符合条件的选用单因素方差分析或t检验，不符合条件的选用秩和检验。P < 0.05为差异有统计学意义。
2. 结果
2.1 旷场实验及糖水偏好实验结果
造模前各组大鼠自发活动总路程长度差异无统计学意义。造模后，与对照组比较，CUS组、si-Atg5组和DAL+si-Atg5组大鼠自发活动总路程长度、糖水消耗量及偏好率降低(P < 0.01)，说明造模成功。干预3周后，与CUS组比较，OAL (0.5 mg/kg)、(1 mg/kg)及(2 mg/kg)组中大鼠自发活动总路程长度、糖水消耗量及偏好率升高(P < 0.05)；si-Atg5组大鼠自发活动总路程长度、糖水消耗量及偏好率降低(P < 0.05)。与OAL (2 mg/kg)组比较，OAL (2 mg/kg)+si-Atg5组大鼠自发活动总路程长度、糖水消耗量及偏好率降低(P < 0.05)。见表 1。
表 1 造模前后各组大鼠旷场实验和糖水偏好实验结果(n=6)Table 1. Open field tests and post-intervention sucrose preference tests on rats (n=6)Group The total moving distance/cm Sucrose preference rate/% Before model After model Before model After model After drug Control 336.4±45.6 489.7±78.2 74.9±2.3 75.0±4.3 75.2±6.9 CUS 328.6±58.3 125.3±54.4** 74.8±3.2 45.7±6.5** 45.8±9.6** OAL (0.5 mg/kg) 331.3±47.6 378.3±96.5# 75.1±2.2 45.8±5.3** 55.2±9.6*，# OAL (1 mg/kg) 352.2±68.2 421.6±87.3# 74.9±2.4 45.6±6.8** 64.3±10.6# OAL (2 mg/kg) 345.7±52.1 463.2±89.7## 75.2±2.7 45.9±5.9** 74.9±9.6## si-Atg5 324.1±66.5 88.6±18.9**，# 74.8±3.4 45.8±4.9** 27.3±5.2**，# OAL+si-Atg5 336.3±72.3 217.6±36.5**，&& 75.0±4.5 45.7±5.2** 55.8±6.9*，& P < 0.05, ** P < 0.01, vs. control group; # P < 0.05, ## P < 0.01, vs. CUS group; & P < 0.05, && P < 0.01, vs. OAL (2 mg/kg) group 2.2 OLA对CUS大鼠海马神经细胞病理的影响
见图 1。与对照组比较，CUS组大鼠海马CA3区发生明显的病理损伤，细胞核浓缩，细胞质减少，细胞核染色加深；与CUS组比较，大鼠经过不同质量浓度的OAL治疗后，细胞核逐渐舒展，细胞形态恢复正常，神经突出逐渐长出；并且随着药物剂量的增加，组织细胞形态越接近对照组；si-Atg5组大鼠海马CA3区病理损伤加重，细胞核浓缩程度、细胞质减少程度、细胞核染色均加重；与OAL (2 mg/kg)组比较，OAL (2 mg/kg)+si-Atg5组大鼠海马区细胞核浓缩程度、细胞质减少程度、细胞核染色均加重。
![]()
图 1 各组大鼠海马CA3区细胞损伤。HE ×400Figure 1. The injury of tissue cells in the CA3 region of hippocampus by group. HE ×400A: Control group; B: CUS group; C: OAL (0.5 mg/kg) group; D: OAL (1.0 mg/kg) group; E: OAL (2.0 mg/kg) group; F: si-Atg5 group; G: OAL (2 mg/kg)+si-Atg5 group2.3 OLA对CUS大鼠海马神经细胞凋亡的影响
见图 2~图 4。与对照组比较，CUS组、OAL(0.5、1 mg/kg)组、si-Atg5组及OAL (2 mg/kg)+si-Atg5组大鼠海马CA3区凋亡细胞百分比(P < 0.01)、cleaved Caspase-3及cleaved Caspase-9表达(P < 0.01)升高。与CUS组比较，OAL (0.5 mg/kg)、(1 mg/kg)及(2 mg/kg)组中凋亡细胞百分比降低(P < 0.01)；OAL (0.5 mg/kg)组cleaved Caspase-3表达无明显变化，cleaved Caspase-9表达降低(P < 0.05)；OAL (1 mg/kg)及(2 mg/kg)组中cleaved Caspase-3、cleaved Caspase-9表达均降低(P < 0.05，P < 0.01)；si-Atg5组凋亡细胞百分比升高(P < 0.01)，cleaved Caspase-3、cleaved Caspase-9表达均升高(P < 0.01)。与OAL (2 mg/kg)组比较，OAL (2 mg/kg)+si-Atg5组凋亡细胞百分比升高(P < 0.01)，cleaved Caspase-3、cleaved Caspase-9表达均升高(P < 0.01)。
![]()
图 2 各组大鼠海马CA3区凋亡细胞。Tunnel染色 ×400Figure 2. Cell apoptosis in the CA3 region of hippocampus by group. Tunnel staining ×400A-G denote the same as fig1![]()
图 3 各组大鼠海马CA3区凋亡细胞百分比Figure 3. The apoptosis rate of cells in the CA3 region of hippocampus by groupA-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group![]()
图 4 各组中cleaved Caspase-3、cleaved Caspase-9的表达Figure 4. The expressions of cleaved Caspase-3 and cleaved Caspase-9 in the CA3 region of hippocampus by groupA-G denote the same as fig 1.*P < 0.05,**P < 0.01,vs. control group; #P < 0.05, ##P < 0.01 ,vs. CUS group; &P < 0.05, &&P < 0.01,vs. OAL (2 mg/kg) group2.4 OLA对CUS大鼠神经细胞炎症反应的影响
见图 5、图 6。与对照组比较，CUS组、OAL(0.5、1 mg/kg)组、si-Atg5组及OAL (2 mg/kg)+si-Atg5组大鼠IL-1β、IL-18血清质量浓度、海马CA3区cleaved Caspase-1、NLRP3表达升高(P < 0.01)。与CUS组比较，OAL (0.5 mg/kg)组IL-1β质量浓度差异无统计学意义，OAL (1 mg/kg)及OAL(2 mg/kg)组中IL-1β质量浓度降低(P < 0.01)；同时，OAL (0.5 mg/kg)、(1 mg/kg)及(2 mg/kg)组中IL-18质量浓度(P < 0.05，P < 0.01)、cleaved Caspase-1、NLRP3表达(P < 0.05，P < 0.01)均降低；si-Atg5组大鼠IL-1β、IL-18血清质量浓度(P < 0.01)、海马CA3区cleaved Caspase-1、NLRP3表达(P < 0.01)升高。与OAL (2 mg/kg)组比较，OAL (2 mg/kg)+si-Atg5组大鼠IL-1β、IL-18血清质量浓度(P < 0.01)、海马CA3区cleaved Caspase-1、NLRP3表达(P < 0.01)升高。
![]()
图 5 各组大鼠血清IL-1β、IL-18质量浓度Figure 5. Serum concentrations of IL-1β and IL-18 by groupA-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group![]()
图 6 各组大鼠海马CA3区组织cleaved Caspase-1、NLRP3的表达Figure 6. The expressions of cleaved Caspase-1 and NLRP3 in the CA3 region of hippocampus by groupA-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group2.5 OLA对CUS大鼠神经细胞自噬的影响
见图 7。与对照组比较，CUS组大鼠海马CA3区LC3Ⅱ/LC3Ⅰ比值、Beclin1表达降低，P62表达升高(P < 0.01)。与CUS组比较，OAL (0.5 mg/kg)、(1 mg/kg)及(2 mg/kg)组中LC3Ⅱ/LC3Ⅰ比值、Beclin1表达升高(P < 0.01)；OAL (0.5 mg/kg)组P62表达差异无统计学意义，(1 mg/kg)及(2 mg/kg)组中P62表达均降低(P < 0.01)；si-Atg5组LC3Ⅱ/LC3Ⅰ比值、Beclin1表达降低(P < 0.05，P < 0.01)，P62表达升高(P < 0.01)。与OAL (2 mg/kg)组比较，OAL (2 mg/kg)+si-Atg5组LC3Ⅱ/LC3Ⅰ比值、Beclin1表达降低(P < 0.01)，P62表达升高(P < 0.01)。
![]()
图 7 各组中LC3、Beclin1、P62的表达Figure 7. The expressions of LC3, Beclin1 and P62 in the CA3 region of hippocampus by groupA-G denote the same as fig1. *P < 0.01, vs. control group; ##P < 0.01, vs. CUS group; &P < 0.01, vs. OAL (2 mg/kg) group3. 讨论
抑郁症可引起患者精神沉郁、快感缺失，改善患者精神状态是抑郁症治疗药物的首要目的[8]。研究发现，脑组织中5-羟色胺水平降低是引起抑郁症发生的重要原因[9]，抗抑郁药OAL是多巴胺/5-羟色胺受体双摄取抑制剂，主要用于治疗双相抑郁症。有报道表明，OAL联合5-羟色胺再摄取抑制剂可显著提高神经元细胞培养上清中5-羟色胺的浓度[10]。本研究发现，0.5、1、2 mg/kg OAL均可显著提高CUS大鼠自发活动总路程、糖水消耗量及偏好率，且随OAL浓度的增加，其对CUS大鼠的作用愈明显。通过对大鼠行为学表现评估表明，OAL可显著改善CUS大鼠的精神状态。
有研究发现，在抑郁症患者脑组织中，海马区神经元细胞发生过度凋亡[11]。海马CA3区是介导应激反应的主要脑区，也是调控人类情绪、记忆及内分泌的重要部位，抑制海马神经元细胞凋亡已成为有效治疗抑郁症的目标[12]。研究表明，OAL对神经元细胞具有显著保护作用，可显著减弱血清戒断诱导的神经元细胞凋亡[13]，降低氧化应激条件下神经元细胞的凋亡水平[14]。在本研究中也发现1、2 mg/kg OAL均可显著降低大鼠海马CA3区凋亡细胞百分比，cleaved Caspase-3、cleaved Caspase-9及cleaved Caspase-1的表达。cleaved Caspase-3、cleaved Caspase-9及cleaved Caspase-1均属于Caspase蛋白水解酶家族，凋亡信号可通过触发Caspase级联反应完成细胞凋亡程序[15]。
炎症反应可显著诱导抑郁症的发生，同时，抑郁症可显著引起脑组织炎性浸润[16]。因此，抑制机体炎症反应是阻碍抑郁症发生发展的有效手段。研究发现，在神经退行性疾病多系统萎缩中，OAL可显著抑制模型小鼠中核因子-κB (NF-κB)的核易位，降低炎症因子IL-1β的表达水平[17]；在应激模型小鼠中，OAL可显著降低小鼠IL-1β和IL-6血清浓度[18]。在本研究中，1、2 mg/kg OAL可显著降低CUS大鼠IL-18及IL-1β血清浓度，同时，0.5、1、2 mg/kg OAL可显著抑制海马CA3区NLRP3炎症小体的表达。NLRP3炎症小体是炎症反应的关键核心，能招募并活化Caspase-1，促进炎症因子IL-1β和IL-18成熟，可作为各种炎症疾病的治疗靶点[19]。NLRP3炎症小体过度激活还可诱导神经元细胞凋亡的发生，抑制NLRP3炎症小体活化可显著减轻CUS模型小鼠抑郁样行为[20]。本实验结果提示，OAL可通过阻碍NLRP3炎症小体激活减轻CUS大鼠的炎症反应。
研究表明，自噬对抑郁症有正向调节作用，如抗抑郁药阿米替林及西酞普兰可提高神经元自噬水平，显著改善模型大鼠抑郁症状[21]；锂可通过诱导自噬显著减少病理朊蛋白的表达水平[22]。本研究中，0.5、1、2 mg/kg OAL均可提高大鼠海马CA3区LC3Ⅱ/LC3Ⅰ比值及Beclin1表达，1、2 mg/kg OAL可降低P62表达。Beclin1是促自噬蛋白，可诱导自噬相关蛋白定位于自噬体膜上，P62是自噬特异性底物，在自噬发生过程中被不断降解，LC3Ⅱ和LC3Ⅰ是LC3的不同表现形式，LC3Ⅰ转变成LC3Ⅱ标志着自噬的形成[23]。本实验结果表明，OAL可显著提高CUS大鼠海马CA3区神经元细胞自噬水平。
据报道，自噬与NLRP3炎症小体密切相关，自噬水平的升高可显著抑制NLRP3炎症小体的激活[24]，NLRP3炎症小体过度活化可引起自噬水平显著降低[25]。Atg5是重要的自噬相关介质，沉默Atg5可诱发NLRP3炎症小体过度活化[21]。本研究利用si-Atg5诱导NLRP3炎症小体过度激活发现，CUS大鼠海马CA3区LC3Ⅱ/LC3Ⅰ比值及Beclin1表达降低，cleaved Caspase-1、cleaved Caspase-3、cleaved Caspase-9表达升高，CUS大鼠自发活动总路程、糖水消耗量及偏好率显著减小。同时，2 mg/kg OAL对CUS大鼠产生的影响被si-Atg5显著减弱。实验结果表明，NLRP3炎症小体过度激活可显著降低CUS大鼠海马CA3区神经元自噬水平，提高炎症反应及细胞凋亡水平，加剧CUS大鼠抑郁症状，并减弱OAL产生的作用。虽然自噬与凋亡在代谢途径和形态学方面有着显著区别，但是他们发挥作用的信号通路却有着交互作用。此外也有研究报道，喜树碱可以通过促进NSCLC自噬而阻止细胞发生程序性死亡[26]。
综上所述，OAL可阻碍NLRP3炎症小体激活，显著减轻CUS大鼠炎症反应，抑制大鼠海马CA3区神经元细胞凋亡，提高神经元细胞自噬水平，改善大鼠的精神状态。同时，NLRP3炎症小体过度激活可显著降低CUS大鼠海马CA3区神经元自噬水平，提高炎症反应及细胞凋亡水平，加剧CUS大鼠抑郁症状，并减弱OAL产生的作用。提示OAL可通过抑制NLRP3炎症小体激活对抑郁症模型大鼠海马神经元产生保护作用，为OAL临床应用提供理论依据。
-
![]()
图 1 各组大鼠海马CA3区细胞损伤。HE ×400
Figure 1. The injury of tissue cells in the CA3 region of hippocampus by group. HE ×400
A: Control group; B: CUS group; C: OAL (0.5 mg/kg) group; D: OAL (1.0 mg/kg) group; E: OAL (2.0 mg/kg) group; F: si-Atg5 group; G: OAL (2 mg/kg)+si-Atg5 group
![]()
图 2 各组大鼠海马CA3区凋亡细胞。Tunnel染色 ×400
Figure 2. Cell apoptosis in the CA3 region of hippocampus by group. Tunnel staining ×400
A-G denote the same as fig1
![]()
图 3 各组大鼠海马CA3区凋亡细胞百分比
Figure 3. The apoptosis rate of cells in the CA3 region of hippocampus by group
A-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group
![]()
图 4 各组中cleaved Caspase-3、cleaved Caspase-9的表达
Figure 4. The expressions of cleaved Caspase-3 and cleaved Caspase-9 in the CA3 region of hippocampus by group
A-G denote the same as fig 1.*P < 0.05,**P < 0.01,vs. control group; #P < 0.05, ##P < 0.01 ,vs. CUS group; &P < 0.05, &&P < 0.01,vs. OAL (2 mg/kg) group
![]()
图 5 各组大鼠血清IL-1β、IL-18质量浓度
Figure 5. Serum concentrations of IL-1β and IL-18 by group
A-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group
![]()
图 6 各组大鼠海马CA3区组织cleaved Caspase-1、NLRP3的表达
Figure 6. The expressions of cleaved Caspase-1 and NLRP3 in the CA3 region of hippocampus by group
A-G denote the same as fig 1.**P < 0.01, vs. control group; #P < 0.05, ##P < 0.01, vs. CUS group; &&P < 0.01, vs. OAL (2 mg/kg) group
![]()
图 7 各组中LC3、Beclin1、P62的表达
Figure 7. The expressions of LC3, Beclin1 and P62 in the CA3 region of hippocampus by group
A-G denote the same as fig1. *P < 0.01, vs. control group; ##P < 0.01, vs. CUS group; &P < 0.01, vs. OAL (2 mg/kg) group
表 1 造模前后各组大鼠旷场实验和糖水偏好实验结果(n=6)
Table 1 Open field tests and post-intervention sucrose preference tests on rats (n=6)
Group The total moving distance/cm Sucrose preference rate/% Before model After model Before model After model After drug Control 336.4±45.6 489.7±78.2 74.9±2.3 75.0±4.3 75.2±6.9 CUS 328.6±58.3 125.3±54.4** 74.8±3.2 45.7±6.5** 45.8±9.6** OAL (0.5 mg/kg) 331.3±47.6 378.3±96.5# 75.1±2.2 45.8±5.3** 55.2±9.6*，# OAL (1 mg/kg) 352.2±68.2 421.6±87.3# 74.9±2.4 45.6±6.8** 64.3±10.6# OAL (2 mg/kg) 345.7±52.1 463.2±89.7## 75.2±2.7 45.9±5.9** 74.9±9.6## si-Atg5 324.1±66.5 88.6±18.9**，# 74.8±3.4 45.8±4.9** 27.3±5.2**，# OAL+si-Atg5 336.3±72.3 217.6±36.5**，&& 75.0±4.5 45.7±5.2** 55.8±6.9*，& P < 0.05, ** P < 0.01, vs. control group; # P < 0.05, ## P < 0.01, vs. CUS group; & P < 0.05, && P < 0.01, vs. OAL (2 mg/kg) group 
下载: 导出CSV
-
[1] SONG W, LI H, GUO T, et al. Effect of affective reward on cognitive event-related potentials and its relationship with psychological pain and suicide risk among patients with major depressive disorder. Suicide Life Threat Behav, 2018 Nov 2[2018-12-19]. https://doi.org/10.1111/sltb.12524.
[2] SOLEM S, HAGEN R, WANG C E, et al. Metacognitions and mindful attention awareness in depression:a comparison of currently depressed, previously depressed and never depressed individuals. Clin Psychol Psychother, 2017, 24(1):94-102. DOI: 10.1002/cpp.1983
[3] DAVIES S J C, MULSANT B H, FLINT A J, et al. SSRI-antipsychotic combination in psychotic depression:Sertraline pharmacokinetics in the presence of olanzapine, a brief report from the STOP-PD study. Hum Psychopharmacol, 2016, 31(3):252-255. https://www.ncbi.nlm.nih.gov/pubmed/27060853
[4] CITROME L. Treatment of bipolar depression:making sensible decisions. CNS Spectr, 2014, 19 Suppl 1:4-11. http://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_PM25407667
[5] TOHEN M, VIETA E, CALABRESE J, et al. Efficacy of olanzapine and olanzapine-fluoxetine combination in the treatment of bipolarⅠ depression. Arch Gen Psychiatry, 2003, 60(11):1079-1088. DOI: 10.1001/archpsyc.60.11.1079
[6] NINAN I, KULKARNI S K. Preferential inhibition of dizocilpine-induced hyperlocomotion by olanzapine. Eur J Pharmacol, 1999, 368(1):1-7. http://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_PM10096763
[7] BANERJEE R, HAZRA S, GHOSH A K, et al. Chronic administration of bacopa monniera increases BDNF protein and mRNA expressions:a study in chronic unpredictable stress induced animal model of depression. Psychiatry Investig, 2014, 11(3):297-306. DOI: 10.4306/pi.2014.11.3.297
[8] WILLIAMS L J, PASCO J A, JACKSON H, et al. Depression as a risk factor for fracture in women: a 10 year longitudinal study. J Affect Disord, 2016, 192:34-40. DOI: 10.1016/j.jad.2015.11.048
[9] CHHIBBER A, WOODY S K, KARIM RUMI M A, et al. Estrogen receptor beta deficiency impairs BDNF-5-HT2A signaling in the hippocampus of female brain: a possible mechanism for menopausal depression. Psychoneuroendo-crinology, 2017, 82:107-116. DOI: 10.1016/j.psyneuen.2017.05.016
[10] ASAOKA N, NAGAYASU K, NISHITANI N, et al. Olanzapine augments the effect of selective serotonin reuptake inhibitors by suppressing GABAergic inhibition via antagonism of 5-HT(6) receptors in the dorsal raphe nucleus. Neuropharmacology, 2015, 95:261-268. DOI: 10.1016/j.neuropharm.2015.03.032
[11] QIN T, FANG F, SONG M, et al. Umbelliferone reverses depression-like behavior in chronic unpredictable mild stress-induced rats by attenuating neuronal apoptosis via regulating ROCK/Akt pathway. Behav Brain Res, 2017, 317:147-156. DOI: 10.1016/j.bbr.2016.09.039
[12] LIU W X, WANG J, XIE Z M, et al. Regulation of glutamate transporter 1 via BDNF-TrkB signaling plays a role in the anti-apoptotic and antidepressant effects of ketamine in chronic unpredictable stress model of depression. Psychopharmacology, 2016, 233(3):405-415. DOI: 10.1007/s00213-015-4128-2
[13] KIM N R, PARK S W, LEE J G, et al. Protective effects of olanzapine and haloperidol on serum withdrawal-induced apoptosis in SH-SY5Y cells. Prog Neuropsychopharmacol Biol Psychiatry, 2008, 32(3):633-642. DOI: 10.1016/j.pnpbp.2007.11.002
[14] WANG H, XU H, DYCK L E, et al. Olanzapine and quetiapine protect PC12 cells from beta-amyloid peptide(25-35)-induced oxidative stress and the ensuing apoptosis. J Neurosci Res, 2005, 81(4):572-580. DOI: 10.1002/jnr.20570
[15] BELMOKHTAR C A, HILLION J, SEGALBENDI-RDJIAN E. Staurosporine induces apoptosis through both caspase-dependent and caspase-independent mechanisms. Oncogene, 2001, 20(26):3354-3362. DOI: 10.1038/sj.onc.1204436
[16] KIECOLT-GLASER J K, DERRY H M, FAGUNDES C P. Inflammation:depression fans the flames and feasts on the heat. Am J Psychiatry, 2015, 172(11):1075-1091. DOI: 10.1176/appi.ajp.2015.15020152
[17] VALERA E, UBHI K, MANTE M, et al. Antidepressants reduce neuroinflammatory responses and astroglial alpha-synuclein accumulation in a transgenic mouse model of multiple system atrophy. Glia, 2014, 62(2):317-337. DOI: 10.1002/glia.22610
[18] THOMAS J, KHANAM R, VOHORA D. Activation of indoleamine 2, 3-dioxygenase pathway by olanzapine augments antidepressant effects of venlafaxine in mice. Psychiatry Res, 2017, 258:444-448. DOI: 10.1016/j.psychres.2017.08.083
[19] ZHONG Z, SANCHEZ-LOPEZ E, KARIN M. Autophagy, NLRP3 inflammasome and auto-inflammatory/immune diseases. Clin Exp Rheumatol, 2016, 34 (4 Suppl 98):12-16. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=3fd5a2cf7068d1e8f956900b74ac6c87
[20] DU R H, TAN J, SUN X Y, et al. Fluoxetine inhibits NLRP3 inflammasome activation: implication in depression. Int J Neuropsychopharmacol, 2016, 19(9).pii: pyw037[2018-10-13].doi: 10.1093/ijnp/pyw037.print2016sep.
[21] ALCOCER-GOMEZ E, CASAS-BARQUERO N, WILLIAMS M R, et al. Antidepressants induce autophagy dependent-NLRP3-inflammasome inhibition in major depressive disorder. Pharmacol Res, 2017, 121:114-121. DOI: 10.1016/j.phrs.2017.04.028
[22] HEISEKE A, AGUIB Y, RIEMER C, et al. Lithium induces clearance of protease resistant prion protein in prion-infected cells by induction of autophagy. J Neurochem, 2009, 109(1):25-34. DOI: 10.1111/j.1471-4159.2009.05906.x
[23] KWON D H, KIM L. pH-dependent regulation of SQSTM1/p62 during autophagy. Autophagy, 2019, 15(1):180-181. DOI: 10.1080/15548627.2018.1532264
[24] SHEN L, YANG Y, OU T, et al. Dietary PUFAs attenuate NLRP3 inflammasome activation via enhancing macrophage autophagy. J Lipid Res, 2017, 58(9):1808-1821. DOI: 10.1194/jlr.M075879
[25] DAI J, ZHANG X, LI L, et al. Autophagy inhibition contributes to ROS-producing NLRP3-dependent inflammasome activation and cytokine secretion in high glucose-induced macrophages. Cell Physiol Biochem, 2017, 43(1):247-256. DOI: 10.1159/000480367
[26] CHIU Y H, HSU S H, HSU H W, et al. Human non-small cell lung cancer cells can be sensitized to camptothecin by modulating autophagy. Int J Oncol, 2018, 53(5):1967-1979.
计量
- 文章访问数: 2498
- HTML全文浏览量: 1068
- PDF下载量: 42
