基于修复肠屏障改善抑郁症小鼠炎症水平的五味子及玫瑰花作用机理研究

Study on the function mechanism of schisandra chinensis and rose based on repairing intestinal barrier and improving inflammation level in mice with depression

研究了五味子及玫瑰花对抑郁模型小鼠行为学和炎症水平的影响,以及其抗抑郁作用和相关机制.在实验中,将36只C57BL/6J小鼠随机分成对照组(Control)、模型组(Model)和五味子及玫瑰花治疗组(S+R),每组12只.除对照组外,模型组和五味子及玫瑰花治疗组均采用慢性不可预知性温和刺激(Chronic unpredictable stress, CUMS)联合孤养制备抑郁模型,连续给予7周刺激,在造模第5周结束后进行行为学检测以判断是否造模成功.在第6周开始,五味子及玫瑰花治疗组给予五味子及玫瑰花灌胃治疗,对照组和模型组给予等量生理盐水.药物干预2周后取材,通过行为学判断小鼠抑郁样行为;Elisa检测小鼠海马5-羟色胺(5-hydroxytryptamine, 5-HT)和脑源性神经营养因子(Brain-derived neurotrophic factor, BDNF)及血清中白细胞介素-6(Interleukin-6,IL-6)、白细胞介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子-α(Tumor Necrosis Factor-α,TNF-α)和内毒素(Lipopolysaccharide, LPS)的水平;Q-PCR检测结肠组织中IL-6、IL-1β、TNF-α和LPS的mRNA相对表达量;HE染色观察结肠组织炎症浸润情况;Western blot检测结肠组织Occludin、claudin1、ZO-1的蛋白表达水平.结果显示,与模型组相比,五味子及玫瑰花治疗组的体重无明显变化(p>0.05);糖水偏好实验治疗组小鼠糖水偏好率显著上升(p<0.01);旷场实验中总路程呈上升趋势(p>0.05),穿越格子数显著上升(p<0.05);强迫游泳中漂浮时间显著缩短(p<0.01);海马中的5-HT、BDNF的含量显著上升(p<0.01);结肠组织炎症浸润程度减轻,同时结肠黏膜屏障相关蛋白Occludin、claudin1、ZO-1表达显著上升(p<0.05),结肠组织中IL-6、IL-1β、TNF-α和LPS的mRNA相对表达量显著下降(p<0.05);血清中的IL-6、IL-1β、TNF-α和LPS水平显著下降(p<0.05).研究表明,五味子及玫瑰花这一药对可改善抑郁模型模型小鼠的抑郁样行为,其机制可能与修复肠黏膜屏障,减少肠源性炎症介质的释放有关.

To observe the effect of Schisandra chinensis and Rose on the behavior and inflammation level of depression model mice,and their antidepressant effects and related mechanisms,36 C57BL/6 mice were randomly divided into control group(Control),model group(Model)and Schisandra chinensis and Rose treatment group(S+R)with 12 mice in each group.In addition to the control group,both model group and the Schisandra chinensis and Rose treatment group used chronic unpredictable mild stimulation(CUMS)combined with solitary cultivation to replicate the depression model.Continuous stimulation for 7 weeks.Behavioral tests were performed at the end of the fifth week to determine whether the model was successful.From the sixth week of stimulation,the S+R group was given rose and schisandra by gavage,and the control group and model group were given the same amount of normal saline.After 2 weeks of drug intervention,the samples were taken.And the depression like behavior of mice was judged to be improved by behavior.Elisa detected 5-hydroxytryptamine(5-HT)and brain-derived neurotrophic factor(BDNF)in hippocampus and Interleukin-6(IL-6),Interleukin-1β(IL-1β),Tumor Necrosis Factor-α(TNF-α)and Lipopolysaccharide(LPS)in serum of mice;Q-PCR detect the relative expression of mRNA of IL-6,IL-1β,TNF-αand LPS in colon tissue;HE staining was used to observe the inflammatory infiltration of colon tissue;Western blot was used to detect the protein expression of Occludin,claudin1,and ZO-1 in colon tissue.The results show that compared with the model group,the body weight of the S+R group had no trend(p>0.05);The sugar preference rate of mice in the treatment group increased significantly(p<0.01);The total distance increased significantly(p>0.05),and the number of crossing grids increased significantly(p<0.05);The floating time was significantly shortened(p<0.01);The contents of 5-HT and BDNF in hippocampus increased significantly(p<0.01);The degree of inflammation infiltration in colon tissue decreased,and the expression of colon mucosal barrier related proteins Occludin,claudin1,ZO-1 increased significantly(p<0.05);The relative expression of mRNA of IL-6,IL-1β,TNF-αand LPS decreased significantly(p<0.05);The levels of IL-6,IL-1β,TNF-αand LPS in serum decreased significantly(p<0.05);Research shows that Schisandra chinensis and Rose can improve the depressive behavior of depression model mice,and its mechanism may be related to repairing intestinal mucosal barrier and reducing the release of intestinal inflammatory mediators.