Abstract：
BACKGROUND: Accurate diagnosis of patients with ulcerative colitis (UC) can reduce their risk of developing colorectal cancer. This study intended to explore whether moxifloxacin, an agent with fluorescence potential, could promote two-photon microscopy (TPM) diagnosis for mice with dextran sodium sulfate (DSS)-induced colitis, which could imitate human UC.
METHODS: 32 Balb/c mice were randomly divided into 4 groups: control, acute colitis, remission colitis and chronic colitis. Fluorescence parameters, imaging performance, and tissue features of different mouse models were compared under moxifloxacin-assisted TPM and label-free TPM.
RESULTS: Excitation wavelength of 720 nm and moxifloxacin labeling time of 2 min was optimal for moxifloxacin-assisted TPM. With moxifloxacin labeling for colonic tissues, excitation power was decreased to 1/10 of that without labeling while fluorescence intensity was increased to 10-fold of that without labeling. Photobleaching was negligible after moxifloxacin labeling and moxifloxacin fluorescence kept stable within 2 h. Compared with the control group, moxifloxacin fluorescence was reduced in the three colitis groups (P < 0.05). Meanwhile, the proportion of enhanced moxifloxacin fluorescence regions was (22.4 ± 1.6)%, (7.7 ± 1.0)%, (13.5 ± 1.7)% and (5.0 ± 1.3)% in the control, acute, remission and chronic groups respectively, with significant reduction in the three colitis groups (P < 0.05). Besides, variant tissue features of experimental colitis models were presented under moxifloxacin-assisted TPM, such as crypt opening, glandular structure, adjacent glandular space and moxifloxacin distribution.
CONCLUSIONS: With unique biological interaction between moxifloxacin and colonic mucosa, moxifloxacin-assisted TPM imaging is feasible and effective for accurate diagnosis of different stages of experimental colitis.
METHODS: 32 Balb/c mice were randomly divided into 4 groups: control, acute colitis, remission colitis and chronic colitis. Fluorescence parameters, imaging performance, and tissue features of different mouse models were compared under moxifloxacin-assisted TPM and label-free TPM.
RESULTS: Excitation wavelength of 720 nm and moxifloxacin labeling time of 2 min was optimal for moxifloxacin-assisted TPM. With moxifloxacin labeling for colonic tissues, excitation power was decreased to 1/10 of that without labeling while fluorescence intensity was increased to 10-fold of that without labeling. Photobleaching was negligible after moxifloxacin labeling and moxifloxacin fluorescence kept stable within 2 h. Compared with the control group, moxifloxacin fluorescence was reduced in the three colitis groups (P < 0.05). Meanwhile, the proportion of enhanced moxifloxacin fluorescence regions was (22.4 ± 1.6)%, (7.7 ± 1.0)%, (13.5 ± 1.7)% and (5.0 ± 1.3)% in the control, acute, remission and chronic groups respectively, with significant reduction in the three colitis groups (P < 0.05). Besides, variant tissue features of experimental colitis models were presented under moxifloxacin-assisted TPM, such as crypt opening, glandular structure, adjacent glandular space and moxifloxacin distribution.
CONCLUSIONS: With unique biological interaction between moxifloxacin and colonic mucosa, moxifloxacin-assisted TPM imaging is feasible and effective for accurate diagnosis of different stages of experimental colitis.
摘要：
背景：对溃疡性结肠炎（UC）患者进行准确诊断可以降低其患大肠癌的风险。本研究旨在探讨莫西沙星是否，具有荧光电位的试剂，可以促进双光子显微镜（TPM）对葡聚糖硫酸钠（DSS）诱导的小鼠结肠炎的诊断，可以模仿人类UC。
方法:32只Balb/c小鼠随机分为4组:对照组,急性结肠炎,缓解性结肠炎和慢性结肠炎。荧光参数,成像性能，在莫西沙星辅助TPM和无标记TPM下比较不同小鼠模型的组织特征。
结果：莫西沙星辅助TPM的激发波长为720nm，莫西沙星标记时间为2分钟是最佳的。用莫西沙星标记结肠组织，激发功率降低到没有标记的1/10，而荧光强度增加到没有标记的10倍。莫西沙星标记后，光漂白可忽略不计，莫西沙星荧光在2小时内保持稳定。与对照组相比,三个结肠炎组的莫西沙星荧光均降低（P<0.05）。同时,增强的莫西沙星荧光区域的比例为（22.4±1.6）％，(7.7±1.0)%,对照组(13.5±1.7)%和(5.0±1.3)%,急性,缓解组和慢性组,3个结肠炎组明显减少(P<0.05)。此外,在莫西沙星辅助TPM下呈现实验性结肠炎模型的变异组织特征，比如地下室开放，腺体结构,邻近腺隙和莫西沙星分布。
结论：莫西沙星与结肠粘膜之间独特的生物学相互作用，莫西沙星辅助TPM显像对实验性结肠炎不同分期的准确诊断具有可行性和有效性。
方法:32只Balb/c小鼠随机分为4组:对照组,急性结肠炎,缓解性结肠炎和慢性结肠炎。荧光参数,成像性能，在莫西沙星辅助TPM和无标记TPM下比较不同小鼠模型的组织特征。
结果：莫西沙星辅助TPM的激发波长为720nm，莫西沙星标记时间为2分钟是最佳的。用莫西沙星标记结肠组织，激发功率降低到没有标记的1/10，而荧光强度增加到没有标记的10倍。莫西沙星标记后，光漂白可忽略不计，莫西沙星荧光在2小时内保持稳定。与对照组相比,三个结肠炎组的莫西沙星荧光均降低（P<0.05）。同时,增强的莫西沙星荧光区域的比例为（22.4±1.6）％，(7.7±1.0)%,对照组(13.5±1.7)%和(5.0±1.3)%,急性,缓解组和慢性组,3个结肠炎组明显减少(P<0.05)。此外,在莫西沙星辅助TPM下呈现实验性结肠炎模型的变异组织特征，比如地下室开放，腺体结构,邻近腺隙和莫西沙星分布。
结论：莫西沙星与结肠粘膜之间独特的生物学相互作用，莫西沙星辅助TPM显像对实验性结肠炎不同分期的准确诊断具有可行性和有效性。
