大蒜素对糖尿病肾病大鼠肾组织细胞焦亡的影响

Effect of Diallyl Trisulfide on Pyroptosis in Renal Tissue of Diabetic Nephropathy Rats

【目的】探讨大蒜素对糖尿病肾病大鼠的干预机制。【方法】将50只大鼠随机分为假手术组,模型组,大蒜素低、中、高剂量组,每组10只。除假手术组外,其余各组大鼠采用高脂饲料喂养结合一次性腹腔注射链脲佐菌素法制备糖尿病肾病模型,假手术组给予普通饲料喂养结合一次性腹腔注射等体积枸橼酸缓冲液。造模成功后,大蒜素高、中、低剂量组分别给予10、20、40 mg·kg^(-1)·d^(-1)大蒜素灌胃,假手术组与模型组给予等体积生理盐水灌胃,每日1次,连续8周。采用苏木素-伊红(HE)、马松(Masson)染色法观察大鼠肾组织病理变化;二氢乙啶(DHE)荧光探针检测肾组织活性氧簇(ROS)水平;蛋白免疫印迹(Western Blot)法检测肾组织含pyrin结构域NOD样受体家族3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)蛋白表达;免疫组织化学法检测肾组织白细胞介素(IL)-1β、IL-18阳性表达。【结果】与假手术组比较,模型组大鼠24 h尿蛋白定量水平升高,肾组织ROS水平及NLRP3、ASC、Caspase-1、IL-1β、IL-18蛋白表达水平升高(均P<0.05),可见肾组织病理损伤。与模型组比较,大蒜素高、中、低剂量组大鼠24 h尿蛋白定量水平下降,肾组织ROS水平及NLRP3、ASC、Caspase-1、IL-1β、IL-18蛋白表达水平下降,且呈剂量依赖性(均P<0.05),可见肾组织病理损伤减轻。【结论】大蒜素可能通过抑制ROS/NLRP3/Caspase-1通路活化减轻糖尿病肾病大鼠肾组织细胞焦亡,从而有效改善糖尿病肾病。

To investigate the intervention mechanism of diallyl trisulfide in rats with diabetic nephropathy.Methods Fifty rats were randomly divided into the sham-operation group,the model group and the diallyl trisulfide low-,medium-and high-dose groups,10 rats in each group.The model of diabetic nephropathy was prepared by feeding the rats with high-fat feed combined with a single intraperitoneal injection of streptozotocin in other groups,except for the sham-operation group,which was fed with fundemental feed combined with a single intraperitoneal injection of equal volume citric acid buffer.After successful modelling,10,20 and 40 mg·kg^(-1)·d^(-1)of diallyl trisulfide were administered by gavage to the high-,medium-and low-dose groups respectively,and an equal volume of normal saline gavage was given to the sham-operation group and model group,once daily for 8 consecutive weeks.The pathological changes of renal tissues of rats were observed by Hematoxylin-eosin(HE)and Masson staining method;the level of reactive oxygen species(ROS)was detected by dihydroethidium(DHE)fluorescence probe;Western Blot was used to detect the protein expression of renal tissue containing pyrin structural domain NOD-like receptor family 3(NLRP3),apoptosis-associated spot-like protein(ASC),and Cysteine aspartate protease 1(Caspase-1);the positive expression of interleukin(IL)-1βand IL-18 in kidney tissues was measured by immunohistochemistry.Results Compared with sham-operation group,24-hour urinary protein amount,ROS level and protein expression levels of NLRP3,ASC,Caspase-1,IL-1βand IL-18 in renal tissues were increased in the model group(all P<0.05),and pathological injury of renal tissue was seen.Compared with model group,24-hour urinary protein amount,ROS level and protein expression levels of NLRP3,ASC,caspase-1,IL-1βand IL-18 in renal tissues of diallyl trisulfide high-,medium-and low-dose groups were decreased in a dose-dependent manner(all P<0.05),and pathological injury of renal tissue was alleviated.Conclusion Diallyl trisulfide may be effective in improving diabetic nephropathy by inhibiting ROS/NLRP3/Caspase-1 pathway activation to attenuate pyroptosis in renal tissue of diabetic nephropathy rats.