基于高通量测序探讨芪藤消浊颗粒对慢性肾炎miRNA表达谱的影响

doi:10.3976/j.issn.1002-4026.20240117

摘要/Abstract

摘要：

基于微小RNA（microRNA，miRNA）高通量测序技术，探讨芪藤消浊颗粒对慢性肾小球肾炎（chronic glomerulonephritis，CGN）大鼠肾小球组织miRNA表达的影响。SD大鼠随机分为正常组、模型组和芪藤消浊颗粒组（n=3），尾静脉注射阿霉素制备CGN模型，连续灌胃芪藤消浊颗粒30 d，采用HE染色观察大鼠肾小球组织病理学影响，miRNA测序技术鉴定差异表达的miRNA，预测其相关mRNA、GO和KEGG富集分析。HE染色结果显示，与正常组相比，模型组大鼠肾小球基底膜增厚，炎性细胞增多，芪藤消浊颗粒干预后，肾小球基底膜和炎性细胞浸润均明显改善。miRNA测序结果显示，与正常组相比，模型组筛选出28个差异表达miRNA；与模型组相比，芪藤消浊颗粒组筛选出28个差异表达miRNA。3组交集共鉴定出12个差异表达miRNA，预测其mRNA，取交集后鉴定出8个核心mRNA。GO主要富集树突棘维持的负电压、谷氨酸门控钙离子通道活性等；KEGG主要参与Rap1、cAMP和RAS信号通路等。qRT-PCR结果显示，3组大鼠肾小球组织差异miRNA的表达水平与测序结果趋势相一致。芪藤消浊颗粒可能通过调控miRNA的表达，进而调控Rap1、cAMP、RAS等信号通路发挥防治CGN的作用。

关键词: 芪藤消浊颗粒, 慢性肾小球肾炎, miRNA, 高通量测序

Abstract:

Using microRNA （miRNA） high-throughput sequencing technology，this study investigatedthe effects of Qiteng Xiaozhuo granules （QTXZG） on miRNA expression in the glomerular tissue of rats with chronic glomerulonephritis （CGN）. Sprague Dawley （SD） rats were randomly divided into three groups： the control group，the model group，and the group receiving QTXZG（n=3）. A CGN model was preparedvia intravenous injection of adriamycin into the tail，followed by 30 days of QTXZG administration via oral gavage.The histopathological effects on rat glomerular tissue were assessed using hematoxylin and eosin staining（HE），and miRNA sequencing was used to identify differentially expressed miRNAs. Subsequent Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were used to predict related mRNAs. HE staining showed that the model group had a thicker glomerular basement membrane and increased inflammatory cell infiltration compared with those in the control group，both of which were significantly improved after undergoing QTXZG treatment. miRNA sequencing revealed 28 differentially expressed miRNAs in the model group compared with the control group，whereas the QTXZG group had 28 differentially expressed miRNAs compared with the model group. A total of 12 differentially expressed miRNAs were identified across the three groups，and after predicting their mRNAs，eight core mRNAs were identified. GO analysis highlighted the negative regulation of dendritic spine maintenance and enrichment in glutamate-gated calcium ion channel activity，whereas KEGG analysis indicated the involvement of the Rap1，cAMP，and RAS signaling pathways. qRT-PCR results confirmed that miRNA expression levels in rat glomerular tissue across the three groups were consistent with the trends observed in the sequencing results. QTXZG regulates the expression of miRNAs，which may，in turn，affect the Rap1，cAMP，and RAS signaling pathways，thus playing a potential role in CGN prevention and treatment.

Key words: Qiteng Xiaozhuo granules, chronic glomerulonephritis, miRNA, high-throughput sequencing technology

中图分类号:

R285

于明哲, 许振, 秦秀娟. 基于高通量测序探讨芪藤消浊颗粒对慢性肾炎miRNA表达谱的影响[J]. 山东科学, 2025, 38(5): 31-39.

YU Mingzhe, XU Zhen, QIN Xiujuan. A high-throughput sequencing-based exploration of the effects of Qiteng Xiaozhuo granules on miRNA expression profiles in chronic glomerulonephritis[J]. Shandong Science, 2025, 38(5): 31-39.

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miRNA名称	扩增子大小/bp	前向引物（5'→3'）	反向引物（5'→3'）
U6	94	CTCGCTTCGGCAGCACA	AACGCTTCACGAATTTGCGT
miR-124-3p		GACTTATAAGGCACGCGGTG	AGTGCAGGGTCCGAGGTATT
miR-124-3p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGGCATT
miR-155-3p		CGCGTATCTCCTACCTGTTAGC	AGTGCAGGGTCCGAGGTATT
miR-155-3p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGTTAAT
miR-216a-5p		GCGAATCTCAGCTGGCAA	AGTGCAGGGTCCGAGGTATT
miR-216a-5p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTCACAG
miR-224-5p		CGCGCAAGTCACTAGTGGTT	AGTGCAGGGTCCGAGGTATT
miR-224-5p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAACGG
miR-1247-3p		CATTACGGGAACGTCGAGAC	AGTGCAGGGTCCGAGGTATT
miR-1247-3p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCTCCA