张绍杰　　汤水福　　陈洪宇　　胡芳玉

[摘要] 目的 分析脾腎阳虚型狼疮性肾炎（LN）患者血浆外泌体miRNA表达谱的变化及探讨其可能存在的意义。方法 选取2015年8～11月于广州中医药大学第一附属医院住院的3例脾肾阳虚型LN患者，并选取健康志愿者3名作为对照组，应用高通量测序技术分析两组患者血浆外泌体miRNA表达谱的差异，应用RT-PCR技术对结果进行验证。 结果 两组的表达谱中，高显著差异的分子包括hsa-miR-127-3p及hsa-miR-4497（P<0.01）;一般显著差异的分子共6个（P<0.05）。PCR验证结果与高通量测序一致。 结论 高显著差异的血浆外泌体miRNA分子可能是脾肾阳虚型LN潜在的生物标志物;差异表达的miRNA分子可能通过促进肾纤维化、介导雌激素作用等参与LN的发生发展。

[关键词] 狼疮性肾炎;脾肾阳虚;miRNA表达谱;高通量测序

[中图分类号] R593.4          [文献标识码] A          [文章编号] 1673-9701（2021）10-0027-04

Analysis and significance of plasma exosome miRNA expression profile in lupus nephritis of spleen-kidney yang deficiency type

ZHANG Shaojie1   TANG Shuifu2   CHEN Hongyu3   HU Fangyu4

1.Department of Nephrology， Dingqiao Hospital of Hangzhou， Hangzhou   310021， China; 2.Department of Nephrology， the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine， Guangzhou   510405， China; 3.Department of Nephrology， Hangzhou Hospital of Traditional Chinese Medicine， Hangzhou   310007， China; 4.Emergency Department， Hangzhou Hospital of Traditional Chinese Medicine， Hangzhou   310007， China

[Abstract] Objective To analyze the changes of plasma exosome miRNA expression profile in patients with lupus nephritis （LN） of spleen-kidney yang deficiency type and explore its possible significance. Methods Three LN patients with spleen-kidney yang deficiency hospitalized in The First affiliated hospital of Guangzhou University of Traditional Chinese Medicine from August to November 2015 were selected. Three healthy volunteers were selected as the control group. High-throughput sequencing technique was used to analyze the difference of plasma exosome miRNA expression profile between the two groups. The results were verified by RT-PCR. Results In the expression profiles of the two groups， the molecules with high significant difference included hsa-miR-127-3p and HSA-MIR-4497 （P<0.01）. Generally， there were 6 molecules with significant differences（P<0.05）. The result of PCR was consistent with that of high-throughput sequencing. Conclusion Plasma exosome miRNA molecules with high significant difference may be potential biomarkers of LN of spleen-kidney yang deficiency type. Differentially expressed miRNA molecules may participate in the occurrence and development of LN via promoting renal fibrosis and mediating estrogenic effects.

[Key words] Lupus nephritis; Spleen-kidney yang deficiency; miRNA expression profile; High throughput sequencing

狼疮性肾炎（Lupus nephritis，LN）是继发性肾脏病变，其发生提示系统性红斑狼疮病情严重。系统性红斑狼疮的病因尚不完全明确，目前认为与遗传因素及性激素等密切相关。其主要发病机制是免疫介导产生的损伤，与多种类型的免疫细胞及细胞因子等相关。

外泌体最早是由Pan等[1]通过电镜发现的直径约40～100 nm、大小均一的脂质双分子层结构囊泡。可由淋巴细胞、树突状细胞等多种细胞分泌，广泛存在于血液、尿液、唾液、脑脊液和胆汁等体液中[2-3]。已证实其在肾脏疾病的发生发展及诊断、治疗方面均有重要的意义[4-5]。外泌体中携带mRNA、miRNA等物质，能传递细胞间的信息，并调控各种生理活动。外泌体中的miRNA分子能调控基因的表达，必然参与疾病的发生发展过程，本研究通过分析脾肾阳虚型LN患者的血浆外泌体miRNA表达谱，拟从分子生物学水平探寻此型LN潜在的发生发展机制，为其诊断及治疗提供帮助。

1 资料与方法