氨磺必利通过TLR/NLR通路对慢性应激抑郁大鼠的改善作用研究

Improvement of amisulpride on chronic stress-depression in rats by TLR/NLR pathway

目的观察氨磺必利对抑郁症大鼠行为的影响,并探究其对大鼠海马组织Toll样受体(TLR)/NOD样受体(NLR)通路相关蛋白表达的影响。方法将SD大鼠随机分为对照组、模型组及低、中、高剂量(0.3、1.2、4.8 g/kg)氨磺必利组,每组10只,除对照组外,其他各组大鼠给予8周慢性温和不可预知应激(CUMS)刺激制备抑郁症大鼠模型,于造模第7~8周,低、中、高剂量氨磺必利组分别ig给予0.3、1.2、4.8 g/kg氨磺必利溶液,对照组、模型组ig等量生理盐水,给药体积10 mL/kg,分别于造模后(CUMS 6周)、给药后(CUMS 8周)采用糖水偏好、强迫游泳、悬尾及旷场实验评估大鼠行为;实验结束后,取各组大鼠海马组织,Werstern blotting法检测TLR2、NLRP1、NLRP3、ASC、TLR4及Caspase-1蛋白表达情况,ELISA法检测白细胞介素(IL)-6、IL-1β、IL-18水平。结果造模后,相较于对照组,模型组及低、中、高剂量氨磺必利组大鼠体质量、糖水偏好指数、旷场中活动距离及站立次数降低,强迫游泳及悬尾静止时间延长(P<0.05)。相较于造模后,给药后模型组大鼠体质量、糖分偏好指数、旷场中活动距离及站立次数降低,强迫游泳及悬尾静止时间延长(P<0.05);低、中、高剂量氨磺必利组大鼠体质量、糖分偏好指数、旷场中活动距离及站立次数增加,强迫游泳及悬尾静止时间缩短(P<0.05)。给药后,相较于对照组,模型组及低、中、高剂量氨磺必利组大鼠体质量、糖分偏好指数、旷场中活动距离及站立次数降低(P<0.05),强迫游泳及悬尾静止时间延长(P<0.05),海马中TLR2、TLR4、NLRP1、NLRP3、ASC、Caspase-1蛋白表达及IL-6、IL-1β、IL-18水平均增高(P<0.05);相较于模型组,低、中、高剂量氨磺必利组大鼠体质量、糖分偏好指数、旷场中活动距离及站立次数增加,且低、中剂量组依次增加,均高于高剂量氨磺必利组(P<0.05),强迫游泳及悬尾静止时间缩短,且低、中剂量组依次缩短,均短于高剂量氨磺必利组(P<0.05),海马中TLR2、TLR4、NLRP1、NLRP3、ASC、Caspase-1蛋白表达及IL-6、IL-1β、IL-18水平均降低,且低、中剂量组依次降低,均低于高剂量氨磺必利组(P<0.05)。结论低剂量氨磺必利可降低抑郁模型大鼠海马中TLR/NLR通路相关蛋白表达量,抑制下游炎性因子释放,从而改善CUMS诱导的大鼠抑郁样行为。

Objective To observe the effect of amisulpride on the behavior of depression rats, and to explore the effect of amisulpride on the expression of Toll like receptor(TLR)/NOD like receptor(NLR) pathway related proteins in the hippocampus of rats. Methods SD rats were randomly divided into control group, model group, and low, medium and high dose(0.3, 1.2, 4.8 g/kg) amisulpride groups, except the control group, rats in the other group were given chronic unpredictable mild stress(CUMS) for 8 weeks to prepare depression rats model. At 7 th—8 th weeks, rats in the low, medium and high dose amisulpride groups were administrated with 0.3, 1.2, 4.8 g/kg amisulpride solutions by ig respectively, rats in the control group and model group were administrated with normal saline by ig, with volume of 10 mL/d. Sugar preference, forced swimming, tail suspension and open field experiments were used to evaluate the behavior of rats after modeling(6 weeks after CUMS) and after administration(8 weeks after CUMS);at the end of the above experiments, the hippocampus of rats were taken, and the expressions of TLR2, NLRP1, NLRP3, ASC, TLR4 and Caspase-1 proteins were detected by Werstern blotting, and the levels of IL-6, IL-1 β and IL-18 were detected by ELISA. Results After molding, compared with those in the control group, the body weight, sugar preference index, open field activity distance and standing times of rats in model group, the low, medium and high dose amisulpride groups were lower, forced swimming time and tail suspension time were longer(P < 0.05). Compared with those after molding, body weight, sugar preference index, activity distance and standing times in open field of rats in model group after administration were lower, forced swimming and tail suspension time were longer(P < 0.05);body weight, sugar preference index, activity distance and standing times in open field of rats in the low, middle and high dose amisulpride groups after administration were higher, forced swimming and tail suspension time were shorter(P < 0.05). After administration, compared with those in the control group, body weight, sugar preference index, open field activity distance and standing times of rats in model group and low, medium and high dose amisulpride groups were lower(P < 0.05), forced swimming and tail suspension time were longer(P < 0.05), the expressions of TLR2, TLR4, NLRP1, NLRP3, ASC, Caspase-1 proteins and the levels of IL-6, IL-1 β, IL-18 in hippocampus were all higher(P < 0.05);compared with those in the model group, the body weight, sugar preference index, open field activity distance and standing times of rats in the low, middle and high dose group were higher, and those in the low and middle dose groups increased in turn, which were higher than those in the high dose group(P < 0.05), the time of forced swimming and tail suspension were shortened, and those in the low and middle dose groups were shorter than those in the high dose group(P < 0.05), the expressions of TLR2, TLR4, NLRP1, NLRP3, ASC, Caspase-1 and the levels of IL-6, IL-1 β, IL-18 in hippocampus were all lower, and those in the low and middle dose groups were lower than those in the high dose group(P < 0.05). Conclusion Low dose of sulfabilide can reduce the expression of TLR/NLR pathway related protein in hippocampus, inhibit the release of downstream inflammatory factors, and improve the depression like behavior of rats induced by CUMS.