探讨电针“人迎”“三阴交”穴通过调控N⁶-甲基腺苷（m⁶A）甲基化修饰减轻盐敏感性高血压（SSHTN）大鼠早期肾损害的分子机制。

将8只SS-13BN大鼠作为对照组。Dahl盐敏感大鼠用含8%NaCl的高盐饲料喂养4周，建立SSHTN大鼠早期肾损害模型，将造模成功的16只Dahl盐敏感大鼠随机分为模型组与电针组（每组8只）。电针组大鼠给予“人迎”“三阴交”电针干预（2/15 Hz，2 mA），20 min/次，1次/d，5 d/周，连续4周。各组大鼠干预前后检测鼠尾收缩压（SBP）及尿微量白蛋白（U-mAlb）；苏木精-伊红（HE）染色法观察大鼠肾脏组织形态；采用RNA-seq与m⁶A-seq联合分析筛选差异m⁶A修饰基因；并运用R语言对交集基因进行基因本体论（GO）功能注解和京都基因与基因组百科全书（KEGG）通路富集分析；实时荧光定量PCR检测大鼠肾组织中甲基转移酶样蛋白3（METTL3）、细胞因子信号转导抑制因子1（SOCS1）、Janus激酶-2（JAK2）、信号转导与转录激活因子3（STAT3）mRNA表达。

干预前，模型组与电针组大鼠SBP及U-mAlb水平均明显高于对照组（模型组：t=19.17、21.96，电针组：t=20.59、21.93，P均<0.001）。经电针干预后，电针组大鼠U-mAlb且较本组干预前明显降低（t=7.33，P<0.001），模型组大鼠SBP、U-mAlb水平均较本组干预前升高（t=23.99、8.22，P均<0.001）；与模型组比较，电针组大鼠SBP、U-mAlb均较明显降低（t=21.53、15.58，P<0.001）。HE染色结果显示，对照组肾小球结构基本正常，模型组肾小球基底膜增厚，炎性浸润明显，电针组较模型组炎性浸润明显减少，肾小球的系膜增生程度减轻。RNA-m⁶A联合测序显示，模型vs对照组肾组织中核心基因SOCS1呈现“Hyper-down”特征（m⁶A修饰升高，mRNA表达降低）；电针组vs模型组肾组织SOCS1基因呈现“Hypo-up”特征（m⁶A修饰降低，mRNA表达升高）。KEGG通路富集分析显示，模型组vs对照组及电针vs模型的差异基因均在JAK/STAT、PI3K-Akt等通路显著富集；GO功能富集结果显示，模型组vs对照组及电针组vs模型组在生物过程中均表现出在蛋白质磷酸化、信号转导及基因表达正向调节条目的高度富集；在分子功能中则富集于蛋白质激酶活性及ATP结合。实时荧光定量PCR结果显示，与对照组比较，模型组大鼠肾组织METTL3、JAK2、STAT3 mRNA相对表达量明显升高（t=5.049、4.605、3.956，P均<0.01），SOCS1 mRNA相对表达量明显降低（t=3.354，P<0.01）。与模型组比较，电针组大鼠肾组织中METTL3、JAK2、STAT3相对表达量明显降低（t=4.199、4.182、4.136，P均<0.05），SOCS1相对表达量明显升高（t=3.335，P<0.05）。

电针“人迎”“三阴交”穴可能通过下调METTL3表达，降低SOCS1 mRNA的m⁶A甲基化水平并抑制JAK/STAT通路的激活，从而发挥对SSHTN大鼠早期肾损害的保护作用。

To investigate the molecular mechanism of electroacupuncture (EA) at "Renying" (ST9) and "Sanyinjiao" (SP6) in alleviating early renal damage in salt-sensitive hypertension (SSHTN) rats by regulating m⁶A methylation modification.

Eight SS-13BN rats were assigned to the control group. Dahl salt-sensitive rats were fed a high-salt diet containing 8% NaCl for 4 weeks to establish an early renal injury model of salt-sensitive hypertensive (SSHTN) rats. Sixteen successfully modeled Dahl salt-sensitive rats were randomly divided into the model group and the electroacupuncture (EA) group, with 8 rats in each group. Rats in the EA group received EA intervention at “Renying” (ST9) and “Sanyinjiao” (SP6) acupoints with parameters of 2/15 Hz, 2 mA, 20 minutes per session, once a day, 5 days a week for 4 consecutive weeks. Systolic blood pressure (SBP) of the rat tail and urinary microalbumin (U-mAlb) were detected in all groups before and after intervention; hematoxylin-eosin (HE) staining was used to observe the morphological changes of rat renal tissues; methylated RNA immunoprecipitation sequencing (m⁶A-seq) combined with RNA sequencing (RNA-seq) was adopted to screen for differentially m⁶A-modified genes; R language was applied to conduct Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the intersecting genes; real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expressions of methyltransferase-like 3 (METTL3), suppressor of cytokine signaling 1 (SOCS1), Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in rat renal tissues.

Before intervention, the levels of SBP and U-mAlb in the model group and EA group were significantly higher than those in the control group (model group: t=19.17, 21.96; EA group: t=20.59, 21.93, all P<0.001). After EA intervention, U-mAlb in the EA group was significantly lower than that before intervention in the same group (t=7.33, P<0.001), while SBP and U-mAlb in the model group were significantly higher than those before intervention in the same group (t=23.99, 8.22, all P<0.001). Compared with the model group, SBP and U-mAlb in the EA group were significantly decreased (t=21.53, 15.58, all P<0.001). HE staining results showed that the glomerular structure was basically normal in the control group; the glomerular basement membrane was thickened with obvious inflammatory infiltration in the model group; while in the EA group, inflammatory infiltration was significantly reduced and mesangial proliferation of glomeruli was alleviated compared with the model group. RNA-m⁶A revealed that the core gene SOCS1 in renal tissues presented a "Hyper-down" characteristic (elevated m⁶A modification and decreased mRNA expression) in the model group vs the control group, and a "Hypo-up" characteristic (decreased m⁶A modification and increased mRNA expression) in the EA group vs the model group. KEGG pathway enrichment analysis indicated that the differentially expressed genes in the model group vs the control group and the EA group vs the model group were significantly enriched in the JAK/STAT, PI3K-Akt and other signaling pathways. GO functional enrichment analysis showed that in the biological process category, the differentially expressed genes in the above two comparisons were highly enriched in items of protein phosphorylation, signal transduction and positive regulation of gene expression; in the molecular function category, they were enriched in protein kinase activity and ATP binding. The results of qRT-PCR showed that compared with the control group, the relative mRNA expressions of METTL3, JAK2 and STAT3 in renal tissues of the model group were significantly increased (t=5.049, 4.605, 3.956, all P<0.01), while the relative mRNA expression of SOCS1 was significantly decreased (t=3.354, P<0.01). Compared with the model group, the relative mRNA expressions of METTL3, JAK2 and STAT3 in renal tissues of the EA group were significantly reduced (t=4.199, 4.182, 4.136, all P<0.05), and the relative mRNA expression of SOCS1 was significantly increased (t=3.335, P<0.05).

EA at ST9 and SP6 acupoints may exert a protective effect on early renal injury in SSHTN rats by down-regulating the expression of METTL3, reducing the m⁶A methylation level of SOCS1 mRNA and inhibiting the activation of the JAK/STAT signaling pathway.