摘要
目的:研究复方高滋斑片(Compound Gaoziban Tablets,CGZBT)对抑郁大鼠的缓解作用和海马NLRP3炎症小体的潜在影响.方法:适应性喂养1周后根据0 weeks糖水偏好率将大鼠随机分为Control组、CUMS组、CGZBT-L组(0.4 g·kg-1)、CGZBT-M组(0.8g·kg-1)、CGZBT-H组(1.6g·kg-1)、Fluoxetine组,每组 12只.SD大鼠造模6周建立慢性不可预知轻度应激(CUMS)抑郁模型,从第5周开始在CUMS造模同时给予相应药物干预2周.最后1天对大鼠进行糖水偏好实验、旷场实验、强迫游泳实验.麻醉后对大鼠海马体、血清进行取材,苏木精-伊红染色法观察海马体CA1区神经元细胞的变化.用ELISA法检测血清中肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)表达水平.Western blot法检测海马体中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)、白细胞介素-1β前体(pro-IL-1β)、IL-1β、白介素-18(IL-18)的表达水平.结果:与CUMS组相比,CGZBT各剂量组旷场移动总距离显著增加(P＜0.01),强迫游泳不动时间减少(P＜0.01或P＜0.05);CGZBT-M组和CGZBT-H组大鼠的糖水偏好率显著增加(P＜0.01).CGZBT可缓解CUMS大鼠海马体CA1区神经元细胞的损伤和降低血清中TNF-α、IL-1β的含量(P＜0.01 或P＜0.05).与CUMS组相比,CGZBT各剂量组大鼠海马中ASC、Caspase-1、IL-1β、的蛋白表达显著下调(P＜0.01或P＜0.05);CGZBT-M组和CGZBT-H组大鼠NLRP3、IL-18蛋白表达显著下调(P＜0.01或P＜0.05).结论:CGZBT可缓解抑郁大鼠的抑郁症状并抑制海马体中NLRP3炎性小体的表达.
Abstract
OBJECTIVE To explore the alleviating effect of Compound Gaoziban Tablets(CGZBT)on depressive rats and examine the potential effects of NLRP3 inflammasome in hippocampus.METHODS After 1-week adaptive feeding,72 Sprague-Dawley(SD)rats were randomized into 6 groups of control,CUMS,CGZBT-L(0.4 g·kg-1),CGZBT-M(0.8 g·kg-1),CGZBT-H(1.6 g·kg-1)and fluoxetine based upon 0 weeks sugar-water preference(n=12 each).A depression model of chronic unpredictable mild stress(CUMS)was established after 6-week modeling.Appropriate pharmacological interventions were administered for 2 weeks starting from Week 5.Sugar preference,open field and forced swimming tests were conducted on the last day.Hippocampal and serum samples were collected after anesthesia.Hematoxylin-easin staining detected the changes of neurons in the CA1 of hippocampal tissue.The expression level of tumor necrosis factor-α(TNF-α)and interleukin-β(IL-1 β)in serum were detected by enzyme-linked inmunosorbent assacy.The expression levels of nod-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate specific proteinase-1(caspase-1),inter leukin-1β precursor(pro-IL-1β),IL-1 β and interleukin-18(IL-18)in hippocampus were detected by Western blot.RESULTS As compared with CUMS group,total distance in open field spiked markedly(P＜0.01)and immobile time of forced swimming declined(P＜0.01 or P＜0.05)in each dose group of CGZBT;sugar preference rate rose obviously in CGZBT-M and CGZBT-H groups(P＜0.01).CGZBT alleviated the injury of neuronal cells in CA1 region of hippocampus and lowered the serum levels of TNF-α and IL-1β(P＜0.01 or P＜0.05).As compared with CUMS group,protein expressions of ASC,caspase-1 and IL-1β were markedly down-regulated in hippocampus of rats in each dose group of CGZBT(P＜0.01 or P＜0.05);and protein expressions of NLRP3 and IL-18 were significantly down-regulated in hippocampus of rats in CGZBT-M and CGZBT-H groups(P＜0.01 or P＜0.05).CONCLUSION CGZBT alleviates depressive symptoms and down-regulates the expression of NLRP3 inflammasome in hippocampus of depressive rats.
NETL
NSTL
万方数据