Zhang Ziyuan, Hu Yaling, Liu Wenyuan, Zhang Xiaodong, Wang Ruihua, Li Hui, Sun Dalin, Fang Jingai
Shanxi Medical University, Taiyuan, People's Republic of China.
Department of Nephrology, First Hospital of Shanxi Medical University, Taiyuan, People's Republic of China.
Diabetes Metab Syndr Obes. 2022 Jul 26;15:2183-2195. doi: 10.2147/DMSO.S368867. eCollection 2022.
To explore the mechanism of Yishen capsule against diabetic nephropathy (DN) based on the analysis of transcriptomics.
SD rats (Male, SPF grade) were randomly divided into four groups, the normal group, the DN group, the Yishen capsule group and the resveratrol group. Urine and renal tissue samples were collected after feeding with physiological saline and above drugs for 8 weeks. 24-hour urine microalbumin protein was detected by ELISA. HE staining and PAS staining were performed on renal tissues. Differential gene expression in renal tissues was analyzed by transcriptome sequencing. The differentially expressed genes were analyzed by GO enrichment and KEGG enrichment, and verified by RT-PCR and immunohistochemistry staining.
The level of 24-hour urinary microalbumin in DN group was increased, while Yishen capsule treatment reversed the increasement of urinary microalbumin. Mesangial cell proliferation, matrix accumulation, edema and vacuolar degeneration of renal tubular epithelial cells and glycogen accumulation were observed in DN group. However, pathological phenotypes mentioned above were alleviated after Yisen capsule administration. This result indicates that Yishen capsule reversed pathological phenotypes of DN in rats. The expression of 261 genes were changed in Yishen capsule group compared with DN group. GO enrichment analysis and KEGG pathway analysis showed that these genes were implicated in pathways, including mineral absorption, adipocytokine signaling pathway, fatty acid biosynthesis, thyroid hormone synthesis, renin-angiotensin system, and NOD-like receptor signaling pathway. Based on previous reported study, the expression of key factors in NOD-like receptor signaling pathway was verified. RT-PCR and immunohistochemistry staining showed that the expression of NLRP3, Caspase-1 and IL-1β in renal tissues of DN group were increased (P < 0.05), which were decreased in Yishen capsule group (P < 0.05).
Yishen capsule reduced microalbuminuria and alleviated pathological changes in DN rats, which may be achieved by regulating NOD-like receptor signaling pathway.
基于转录组学分析探讨益肾胶囊抗糖尿病肾病（DN）的作用机制。
将雄性SPF级SD大鼠随机分为四组，即正常组、DN组、益肾胶囊组和白藜芦醇组。用生理盐水及上述药物喂养8周后收集尿液和肾组织样本。采用酶联免疫吸附测定法检测24小时尿微量白蛋白。对肾组织进行苏木精-伊红（HE）染色和过碘酸雪夫（PAS）染色。通过转录组测序分析肾组织中的差异基因表达。对差异表达基因进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）富集分析，并通过逆转录-聚合酶链反应（RT-PCR）和免疫组织化学染色进行验证。
DN组24小时尿微量白蛋白水平升高，而益肾胶囊治疗可逆转尿微量白蛋白的升高。DN组可见系膜细胞增殖、基质积聚、肾小管上皮细胞水肿和空泡变性以及糖原积聚。然而，给予益肾胶囊后上述病理表型得到缓解。这一结果表明益肾胶囊可逆转大鼠DN的病理表型。与DN组相比，益肾胶囊组有261个基因的表达发生了变化。GO富集分析和KEGG通路分析表明，这些基因涉及矿物质吸收、脂肪细胞因子信号通路、脂肪酸生物合成、甲状腺激素合成、肾素-血管紧张素系统和NOD样受体信号通路等途径。基于先前报道的研究，对NOD样受体信号通路中的关键因子表达进行了验证。RT-PCR和免疫组织化学染色显示，DN组肾组织中NLRP3、半胱天冬酶-1（Caspase-1）和白细胞介素-1β（IL-1β）的表达增加（P<0.05），而在益肾胶囊组中则降低（P<0.05）。
益肾胶囊可降低DN大鼠的微量白蛋白尿并减轻其病理变化，这可能是通过调节NOD样受体信号通路实现的。
