目的：基于转录组学探讨和血明目片改善糖尿病视网膜病变(DR)的作用机制。

方法：将3日龄(3 dpf)斑马鱼幼鱼持续暴露于130 mmol/L葡萄糖溶液3 d构建DR模型，随机分为对照组(给予养殖水)、模型组(130 mmol/L葡萄糖)、低剂量治疗组(130 mmol/L葡萄糖+7.5 mg/L和血明目片)、高剂量治疗组(130 mmol/L葡萄糖+75 mg/L和血明目片)，持续干预至6 dpf。通过体视显微镜观察斑马鱼眼睛面积、眼睛长度和体长，采用苏木精-伊红(HE)染色法观察视网膜形态，荧光显微镜下观察其视网膜血管直径，通过RNA高通量测序(RNA-seq)技术筛选差异表达基因(DEGs)，进一步阐释和血明目片改善斑马鱼DR的分子机制，并用qRT-PCR检测基因的表达以验证测序结果的准确性。

结果：HE染色显示模型组节细胞层(GCL)细胞排列紊乱、间隙增宽、内核层(INL)增厚，和血明目片干预后GCL层细胞紊乱有所缓解，INL厚度降低； 视网膜血管直径定量分析显示，模型组斑马鱼视网膜血管直径较对照组显著增加，和血明目片干预后显著逆转，其中高剂量和血明目片治疗组效果更显著(P<0.05)； 转录组分析共鉴定出1 470个逆转DEGs，主要富集于AMPK信号通路、Foxo信号通路、视网膜发育和紧密连接等过程； 相关性分析表明qRT-PCR结果与转录组学结果高度一致(R²=0.8571，P<0.05)。

结论：和血明目片可能通过调控vsx1、pde6c、arr3a、plk1、fbp1b、foxo1a、pcna和cdk1等核心靶点，并通过对相机型眼的视网膜发育、视觉感知、细胞骨架组织、紧密连接、AMPK信号通路、Foxo信号通路等过程的协同调控改善DR视网膜血管微循环障碍。

METHODS: Zebrafish DR models were established by 3-day glucose induction(130 mmol/L)starting at 3 days post-fertilization(dpf). Larvae were randomized into four groups: control group(CG; aquaculture water), model group(MG; 130 mmol/L glucose), low-dose HXMMT treatment group(L-HX; 130 mmol/L glucose +7.5 mg/L HXMMT), and high-dose HXMMT treatment group(H-HX; 130 mmol/L glucose +75 mg/L HXMMT), with a 3-day intervention period until 6 dpf. The area and length of eyes, and body length of zebrafish were observed by stereomicroscopy, retinal morphology was observed by hematoxylin-eosin staining(HE), and retinal vessel diameter was observed under fluorescence microscope. Differentially expressed genes(DEGs)were identified by RNA-sequencing(RNA-seq)technology to further elucidate the molecular mechanism of HXMMT in improving DR in zebrafish, and the sequencing accuracy was validated through quantitative real-time polymerase chain reaction(qRT-PCR).

RESULTS: HE staining demonstrated that the intervention with HXMMT significantly improved the disordered cell arrangement, widened gaps, and thickened inner nuclear layer(INL)in ganglion cell layer GCL); retinal vascular diameter quantification revealed that the retinal vessel diameter of the MG significantly increased compared with the CG, and it was significantly changed after the intervention of HXMMT, with significant efficacy in the H-HX(P<0.05); transcriptomics profiling identified 1 470 reversed DEGs, predominantly enriched in the AMPK signaling pathway, FoxO signaling pathway, retinal developmental processes, and tight junction regulation. Technical validation confirmed strong correlation between qRT-PCR and RNA-seq data(R²=0.8571, P<0.05).

CONCLUSION: HXMMT may improve retinal vascular microcirculation disorders in DR by regulating core targets including vsx1, pde6c, arr3a, plk1, fbp1b, foxo1a, pcna, and cdk1, as well as synergistically modulating processes such as retinal development in camera-type eyes, visual perception, microtubule cytoskeletal organization, tight junctions, and the AMPK signaling pathway, Foxo signaling pathway.