目的 探讨菖蒲郁金汤通过调节小胶质细胞(MG)极化对3，3'-亚氨基二丙腈(IDPN)诱导的抽动-秽语综合征(TS)模型大鼠细胞因子及神经递质的影响。方法 选取雄性SD大鼠40只，按照随机数字表法分为空白组、模型组、硫必利组和菖蒲郁金汤组，各10只。使用IDPN腹腔注射制备TS模型，采用运动行为评分表检测造模是否成功。硫必利组给予硫必利混悬液，菖蒲郁金汤组给予菖蒲郁金汤药液，按15 mL/(kg·d)灌胃，空白组和模型组给予等体积生理盐水。造模成功后各组分别取5只大鼠，处死并取材;各组剩余5只大鼠干预28 d后全部处死并取材。观察各组大鼠纹状体组织形态的病理改变、MG的极化分型及纹状体细胞因子[白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、肿瘤坏死因子α(TNF-α)和神经递质多巴胺(DA)、多巴胺受体(DR)、γ-氨基丁酸A型受体(GABAAR)]的水平变化。结果 干预前，各组间行为学评分比较，差异均无统计学意义(P＞0。05)，具有可比性;干预后，硫必利组和菖蒲郁金汤组的刻板行为评分和运动行为评分均较模型组显著降低，差异均有统计学意义(P＜0。05)，菖蒲郁金汤组的刻板行为评分与硫必利组比较，差异有统计学意义(P＜0。05)，而2组的运动行为评分比较，差异无统计学意义(P＞0。05)。TS大鼠纹状体组织病理切片显示，干预前空白组大鼠纹状体组织排列整齐且紧密、细胞大小适中，神经元核仁较大而圆，形态规则，细胞核与间质轮廓清晰;模型组纹状体组织排列稀疏且紊乱，神经元部分发生变性、坏死，细胞核固缩，胞质肿胀。干预后，硫必利组纹状体神经细胞数量、体积和形态变化与模型组比较无明显区别;菖蒲郁金汤组纹状体神经细胞数量有所增加，胞体变圆，神经元排列紊乱程度及胞质肿胀程度减轻，皱缩坏死减少。干预前，与空白组比较，TS模型组、硫必利组和菖蒲郁金汤组大鼠纹状体中促炎(M1)型小胶质细胞比抗炎(M2)型小胶质细胞比例(M1/M2)显著升高，差异有统计学意义(P＜0。05)，而精氨酸酶1(Arg1)标记的M2型小胶质细胞比例无明显变化(P＞0。05);此时M1型小胶质细胞多于M2型，占主导地位。干预后，与空白组比较，模型组大鼠纹状体M1型比例显著升高(P＜0。01);与模型组比较，硫必利组和菖蒲郁金汤组M1型小胶质细胞比例显著降低，M2型小胶质细胞比例显著升高，差异均有统计学意义(P＜0。01)，此时M2型小胶质细胞多于M1型，占主导地位。干预前，与空白组比较，模型组、硫必利组和菖蒲郁金汤组中IL-1β、IL-6、TNF-α水平均升高，差异均有统计学意义(P＜0。05)，IL-10、DA、DR、GAB A AR水平均降低，差异均有统计学意义(P＜0。05);干预后，与空白组比较，模型组IL-1β、IL-6、TNF-α水平显著升高，IL-10、DA、DR、GABAAR水平显著较低，差异均有统计学意义(P＜0。05);与模型组比较，硫必利组和菖蒲郁金汤组IL-1β、IL-6、TNF-α水平均显著降低，IL-10、DA、DR、GABAAR水平均显著升高，差异有统计学意义(P＜0。05)。结论 菖蒲郁金汤可能是通过调节MG极化并维持DR-GABAAR相互作用的稳态，进一步改善TS免疫炎症状态，调控神经递质平衡，从而发挥抗抽动作用。
Experimental study on the anti-twitching effect of Changpu Yujin Tang by regulating microglia and neurotransmitter balance in a rat model of Tourette syndrome
Objective To investigate the effects of calamus and turmeric root-tuber decoction on cytokines and neurotransmitters in 3,3'-iminodipropionitrile(IDPN)-induced TS rat model by regulating microglia(MG)polari-sation.Methods Forty male SD rats were selected and divided into a blank group,a model group,a thiabendazole group,and a calamus and turmeric root-tuber decoction group,10 each according to the random number table meth-od.The TS model was prepared by intraperitoneal injection using IDPN,and the success of modelling was detected using the motor behaviour scale.The thiabendazole group was given thiabendazole suspension,the Changpu Yujin Tang(CYT)group was given Changpu Yujin Tang medicinal solution by gavage at the amount of 15 mL/(kg·d),and the blank group and model group were given equal volumes of saline.After successful modelling,5 rats were taken from each group,executed and sampled;the remaining 5 rats in each group were all executed and sampled 28 days after the intervention.Pathological changes in the histomorphology of the striatum,polarised typing of MG and changes in the levels of striatal cytokines[interleukin-1β(IL-1 β),IL-6,IL-10,tumour necrosis factor α(TNF-α)and the neurotransmitters dopamine(DA),dopamine receptor(DR),γ-aminobutyric acid type A receptor(GABAAR)]were observed in the rats in each group.Results Before the intervention,the comparison of behav-ioural scores between the groups was comparable with no statistically significant difference(P＞0.05);after the in-tervention,the stereotyped behavioural scores and motor behavioural scores of the thiabendazole group and the CYT group were significantly reduced compared with those of the model group,and the differences were all statistically significant(P＜0.05),and the difference of the stereotyped behavioural scores of the CYT group was statistically significant when compared with that of the thiabendazole group(P＜0.05),while the difference between the motor behaviour scores of the two groups was not statistically significant(P＞0.05).Histopathological sections of the stria-tum of rats in the TS group showed that the striatum of rats in the blank group was neatly arranged and tightly packed,with moderate cell sizes,large and rounded neuronal nuclei,regular morphology,and clear outlines of the nuclei and the interstitial matrix before the intervention,while that of the model group was sparsely arranged and disorganized,with neurons partly degenerating,necrotic,with nuclei and interstitial matrix.In the model group,the striatal tissue was sparsely arranged and disorganised,and neurons were partially degenerated,necrotic,with nuclei consolidated and cytoplasm swollen.After the intervention,there was no significant difference in the number,volume and morphology of striatal nerve cells in the thiabendazole group compared with the model group;the number of stri-atal nerve cells in the CYT group increased,the cytosol became rounded,the degree of neuronal arrangement disor-der and cytoplasmic swelling was reduced,and the degree of wrinkled and necrotic cells was reduced.Before inter-vention,compared with the blank group,the ratio of pro-inflammatory(Ml)microglia to anti-inflammatory(M2)microglia(M1/M2)in the striatum of rats in the TS model group,the thiabendazole group,and the CYT group was significantly higher,and the difference was statistically significant(P＜0.05),whereas the ratio of Arg1-labelled M2-type microglia did not change significantly(P＞0.05);at this time,there were more M1-type microglia than M2-type microglia,cells were more predominant than M2 type.Intervention,compared with the blank group,the proportion of M1 in the striatum of rats in the model group was significantly increased(P＜0.01);compared with the model group,the proportion of M1-type microglia in the thiabendazole group and the CYT group was significantly de-creased,and the proportion of M2-type microglia was significantly increased,and the differences were statistically significant(P＜0.01),at this time,there were more M2-type microglia than M1-type,which were predominant.Be-fore intervention,compared with the blank group,the levels of IL-1β,IL-6,and TNF-α in the model group,the thia-bendazole group,and the CYT group were all elevated,and the differences were all statistically significant(P＜0.05),and the levels of IL-10,DA,DR,and GAB AAR were all reduced,and the differences were all statistically significant(P＜0.05);after intervention,compared with the blank group,in the model group,the levels of IL-1β,IL-6,TNF-α levels were significantly higher,and the difference was statistically significant(P＜0.05),and IL-10,DA,DR,and GABA AR levels were significantly lower,and the difference was statistically significant(P＜0.05);com-pared with the model group,both the thiabendazole group and the CYT group had significantly lower IL-1 β,IL-6,and TNF-α levels,and IL-10,DA,and DR,GAB AAR levels were significantly increased,and the difference was sta-tistically significant(P＜0.05).Conclusion Changpu Yujin Tang may exert its anti-twitching effect by regulating MG polarisation and maintaining the homeostasis of DR-GABAAR interaction,further improving the TS immuno-in-flammatory state,and regulating the neurotransmitter balance.
Tourette syndromeChangpu Yujin Tangmicroglia polarisationcytokinesneurotransmittersexper-imental study
万方数据
维普
