Abstract：
BACKGROUND: Altered gut metabolites, especially short-chain fatty acids (SCFAs), in feces and plasma are observed in patients with Parkinson\'s disease (PD).
OBJECTIVE: We aimed to investigate the colonic expression of two SCFA receptors, free fatty acid receptor (FFAR)2 and FFAR3, and gut barrier integrity in patients with PD and correlations with clinical severity.
METHODS: In this retrospective study, colonic biopsy specimens were collected from 37 PD patients and 34 unaffected controls. Of this cohort, 31 participants (14 PD, 17 controls) underwent a series of colon biopsies. Colonic expression of FFAR2, FFAR3, and the tight junction marker ZO-1 were assayed by immunofluorescence staining. The You Only Look Once (version 8, YOLOv8) algorithm was used for automated detection and segmentation of immunostaining signal. PD motor function was assessed with the Movement Disorder Society (MDS)-Unified Parkinson\'s Disease Rating Scale (UPDRS), and constipation was assessed using Rome-IV criteria.
RESULTS: Compared with controls, PD patients had significantly lower colonic expression of ZO-1 (p < 0.01) and FFAR2 (p = 0.01). On serial biopsy, colonic expression of FFAR2 and FFAR3 was reduced in the pre-motor stage before PD diagnosis (both p < 0.01). MDS-UPDRS motor scores did not correlate with colonic marker levels. Constipation severity negatively correlated with colonic ZO-1 levels (r = -0.49, p = 0.02).
CONCLUSIONS: Colonic expression of ZO-1 and FFAR2 is lower in PD patients compared with unaffected controls, and FFAR2 and FFAR3 levels decline in the pre-motor stage of PD. Our findings implicate a leaky gut phenomenon in PD and reinforce that gut metabolites may contribute to the process of PD.
OBJECTIVE: We aimed to investigate the colonic expression of two SCFA receptors, free fatty acid receptor (FFAR)2 and FFAR3, and gut barrier integrity in patients with PD and correlations with clinical severity.
METHODS: In this retrospective study, colonic biopsy specimens were collected from 37 PD patients and 34 unaffected controls. Of this cohort, 31 participants (14 PD, 17 controls) underwent a series of colon biopsies. Colonic expression of FFAR2, FFAR3, and the tight junction marker ZO-1 were assayed by immunofluorescence staining. The You Only Look Once (version 8, YOLOv8) algorithm was used for automated detection and segmentation of immunostaining signal. PD motor function was assessed with the Movement Disorder Society (MDS)-Unified Parkinson\'s Disease Rating Scale (UPDRS), and constipation was assessed using Rome-IV criteria.
RESULTS: Compared with controls, PD patients had significantly lower colonic expression of ZO-1 (p < 0.01) and FFAR2 (p = 0.01). On serial biopsy, colonic expression of FFAR2 and FFAR3 was reduced in the pre-motor stage before PD diagnosis (both p < 0.01). MDS-UPDRS motor scores did not correlate with colonic marker levels. Constipation severity negatively correlated with colonic ZO-1 levels (r = -0.49, p = 0.02).
CONCLUSIONS: Colonic expression of ZO-1 and FFAR2 is lower in PD patients compared with unaffected controls, and FFAR2 and FFAR3 levels decline in the pre-motor stage of PD. Our findings implicate a leaky gut phenomenon in PD and reinforce that gut metabolites may contribute to the process of PD.
摘要：
背景：改变肠道代谢产物，特别是短链脂肪酸（SCFA），在帕金森病（PD）患者的粪便和血浆中观察到。
目的：我们旨在研究两种SCFA受体的结肠表达，游离脂肪酸受体（FFAR）2和FFAR3，与PD患者的肠屏障完整性以及与临床严重程度的相关性。
方法：在这项回顾性研究中，收集了37例PD患者和34例未受影响的对照组的结肠活检标本.在这个队列中，31名参与者(14名PD，17名对照）接受了一系列结肠活检。通过免疫荧光染色检测FFAR2、FFAR3和紧密连接标记物ZO-1的结肠表达。YouOnlyLookOnce（版本8，YOLOv8）算法用于免疫染色信号的自动检测和分割。使用运动障碍协会（MDS）-统一帕金森病评定量表（UPDRS）评估PD运动功能，便秘采用Rome-IV标准进行评估.
结果：与对照组相比，PD患者的结肠ZO-1（p<0.01）和FFAR2（p=0.01）表达显着降低。在连续活检中，在PD诊断前的运动前阶段，FFAR2和FFAR3的结肠表达降低（均p<0.01）。MDS-UPDRS运动评分与结肠标志物水平无关。便秘严重程度与结肠ZO-1水平呈负相关(r=-0.49,p=0.02)。
结论：与未受影响的对照组相比，PD患者中ZO-1和FFAR2的结肠表达较低，在PD运动前阶段，FFAR2和FFAR3水平下降。我们的发现暗示PD中存在漏肠现象，并加强了肠道代谢产物可能有助于PD的过程。
目的：我们旨在研究两种SCFA受体的结肠表达，游离脂肪酸受体（FFAR）2和FFAR3，与PD患者的肠屏障完整性以及与临床严重程度的相关性。
方法：在这项回顾性研究中，收集了37例PD患者和34例未受影响的对照组的结肠活检标本.在这个队列中，31名参与者(14名PD，17名对照）接受了一系列结肠活检。通过免疫荧光染色检测FFAR2、FFAR3和紧密连接标记物ZO-1的结肠表达。YouOnlyLookOnce（版本8，YOLOv8）算法用于免疫染色信号的自动检测和分割。使用运动障碍协会（MDS）-统一帕金森病评定量表（UPDRS）评估PD运动功能，便秘采用Rome-IV标准进行评估.
结果：与对照组相比，PD患者的结肠ZO-1（p<0.01）和FFAR2（p=0.01）表达显着降低。在连续活检中，在PD诊断前的运动前阶段，FFAR2和FFAR3的结肠表达降低（均p<0.01）。MDS-UPDRS运动评分与结肠标志物水平无关。便秘严重程度与结肠ZO-1水平呈负相关(r=-0.49,p=0.02)。
结论：与未受影响的对照组相比，PD患者中ZO-1和FFAR2的结肠表达较低，在PD运动前阶段，FFAR2和FFAR3水平下降。我们的发现暗示PD中存在漏肠现象，并加强了肠道代谢产物可能有助于PD的过程。
