【摘要】 【目的】探究大蒜素对2型糖尿病肾病（DN）的治疗作用及机制。【方法】将27只雄性SD大鼠随机分为假手术组、模型组与大蒜素组，每组9只。模型组与大蒜素组大鼠构建DN模型。建模成功后，分组灌胃干预。检测肾功能指标、氧化应激指标，采用荧光探针法检测大鼠肾组织中活性氧（ROS）水平，通过碘酸-希夫（PAS）染色、苏木精-伊红（HE）染色观察肾组织病理形态，采用透射电镜观察肾组织细胞超微结构，Western Blot法观察肾组织焦亡相关蛋白[气体衍生蛋白E(GSDME)、NLR家族pyrin结构域蛋白1(NLRP1)、NLR家族pyrin结构域蛋白3(NLRP3)、半胱氨酸天冬氨酸特异性蛋白酶1(Caspase1)、热休克蛋白家族A成员1B(HSPA1B)]水平，实时定量逆转录聚合酶链式反应（RT-qPCR）法检测肾组织细胞色素P450家族1亚家族B成员1反义RNA1(CYP1B1-AS1) mRNA表达水平，RNA免疫共沉淀（RIP）法验证HSPA1B与CYP1B1-AS1互作情况。【结果】与假手术组比较，模型组肾功能指标、氧化应激水平明显异常，肾组织炎症浸润、糖原沉积及细胞出现焦亡特征，肾组织ROS水平升高，焦亡相关蛋白（GSDME、NLRP1、NLRP3、Caspase1、HSPA1B）及CYP1B1-AS1 mRNA表达水平升高（P<0.05）；与模型组比较，治疗组大鼠肾功能指标、氧化应激水平显著改善，肾组织炎症浸润、糖原沉积及细胞焦亡特征减轻，肾组织ROS水平降低，焦亡相关蛋白（GSDME、NLRP1、NLRP3、HSPA1B、Caspase1）及CYP1B1-AS1 mRNA表达水平下调（P<0.05）。RIP实验证实HSPA1B与CYP1B1-AS1存在互作。【结论】大蒜素可通过调控CYP1B1-AS1/HSPA1B/ROS通路，减轻肾组织氧化应激损伤与细胞焦亡，进而缓解DN进展。更多还原

【Abstract】 Objective To investigate the therapeutic effects and underlying mechanism of allicin on type 2 diabetic nephropathy(DN). Methods Twenty-seven male SD rats were randomly divided into a sham-operated group,a model group, and an allicin group, with 9 rats in each group. The DN model was established in rats from the model and allicin groups. After successful modeling,the groups received respective interventions. Renal function indicators and oxidative stress markers were assessed. Reactive oxygen species(ROS) level in rat kidney tissues was detected using a fluorescent probe method. Renal histopathology was observed by periodic acid-Schiff(PAS) staining and hematoxylin-eosin(HE) staining, and the ultrastructure of renal tissue cells was examined by transmission electron microscopy. Western Blot analysis was employed to detect pyroptosis-related proteins [gasdermin E(GSDME),NLR family pyrin domain containing 1(NLRP1),NLR family pyrin domain containing 3(NLRP3), caspase-1, and heat shock protein family A member 1B(HSPA1B)] in renal tissues. Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) was used to measure the mRNA expression level of cytochrome P450 family 1 subfamily B member 1 antisense RNA 1(CYP1B1-AS1). RNA immunoprecipitation(RIP) assay was performed to validate the interaction between HSPA1B and CYP1B1-AS1. Results Compared with the sham-operated group,the model group exhibited significantly abnormal renal function indicators and oxidative stress levels,marked inflammatory infiltration,glycogen deposition,and characteristic features of pyroptosis in renal tissues. ROS level in renal tissues was significantly elevated,accompanied by increased protein expression levels of pyroptosis-related markers(GSDME, NLRP1, NLRP3, Caspase1, HSPA1B) and upregulation of CYP1B1-AS1 mRNA expression(P<0.05). In contrast, compared with the model group, the allicin-treated group demonstrated significantly improved renal function and oxidative stress parameters, alleviated renal inflammatory infiltration, glycogen deposition, and pyroptotic features. ROS level in renal tissues was notably reduced, and the expression levels of pyroptosis-related proteins(GSDME, NLRP1, NLRP3, HSPA1B,Caspase1) and CYP1B1-AS1 mRNA were significantly downregulated(P<0.05). RIP assay confirmed the interaction between HSPA1B and CYP1B1-AS1. Conclusion Allicin can mitigate the progression of DN by regulating the CYP1B1-AS1/HSPA1B/ROS pathway, and alleviating oxidative stress damage and pyroptosis in renal tissues.更多还原