目的 探讨吡咯喹啉醌（pyrroloquinoline quinone,PQQ）是否对糖尿病导致的肝损伤具有抑制作用。方法 C57BL/6小鼠随机分为5组（n = 10）,包括对照组、DM组、DM+ PQQ低剂量组、DM + PQQ中剂量组和DM + PQQ高剂量组,除对照组以外的小鼠通过腹腔注射链脲佐菌素（STZ）诱导1型糖尿病,模型成功后每日灌胃给予低剂量（10 mg/kg)、中剂量（20 mg/kg）、高剂量（40 mg/kg）PQQ进行干预,持续12 w。检测小鼠体重和空腹血糖变化,HE染色检测肝脏组织病理学改变,试剂盒检测肝损伤相关血生化指标及氧化应激指标,Western blot检测核苷酸脱脂寡聚结构域样吡啶结构域蛋白3（nucleotide binding oligomer-rization domain-like pyrin domain containing protein 3,NLRP3）炎症小体相关蛋白表达变化。结果 40 mg/kg PQQ干预显著降低空腹血糖水平,降低血清碱性磷酸酶（alkaline phosphatase,ALP）和丙氨酸氨基转移酶（alanine aminotransferase,ALT）水平,减少血清中炎症相关因子白细胞介素1β（interleukin-1 beta,IL-1β）、白细胞介素6（interleukin-6,IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）含量,显著提高肝脏组织SOD活性和GSH含量。组织病理学分析也显示PQQ能够减轻肝脏损伤,Western blot分析显示PQQ显著减少NLRP3炎症小体信号通路NLRP3、硫氧还蛋白互作蛋白（thioredoxin interacting protein,TXNIP）、凋亡相关斑点样蛋白（apoptosis - associated speck - like protein containing a CARD,ASC）、半胱氨酸天冬氨酸蛋白酶1（cysteinyl aspartate specific proteinase-1,Caspase-1）、Caspase-1 cut、IL-1β以及白细胞介素18（interleukin-18,IL-18）蛋白表达。结论 PQQ对糖尿病导致的肝脏损伤具有明显的抑制作用,其机制可能是增强抗氧化能力及减少NLRP3炎症小体信号通路蛋白表达。
Abstract
Objective To elucidate the potential protective effects of PQQ against diabetic liver damages and its underlying mechanisms. Methods C57BL/6 mice were randomly divided into 5 groups (n=10), including control group, DM group, DM+ PQQ low dose group, DM+ PQQ medium dose group and DM+ PQQ high dose group. Streptozotocin (STZ) was used by intraperitoneal injection to develop type 1 diabetes. Following successful modeling, mice were treated with daily oral administration of low dose (10 mg/kg), medium dose (20 mg/kg), and high dose (40 mg/kg) of PPQ for a duration of 12 weeks. The changes of body weight and fasting blood glucose were recorded. Histopathological alterations of liver tissue were examined using HE staining. Additionally, liver damage-associated blood biochemical markers and antioxidant enzyme activities were assessed utilizing commercial reagent kits. The expressions of (nucleotide binding oligomer-rization domain-like pyrin domain containing protein 3 (NLRP3) inflammasome-related proteins were detected by Western blot analysis. Results Treatment with a dosage of 40 mg/(kg·d ) PQQ significantly decreased fasting blood glucose levels and liver injury-related enzymes alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activites, reduced serum levels of inflammatory factors interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and enhanced the activity of antioxidant enzyme SOD and GSH content in the liver. Histopathological analysis also demonstrated that PQQ exhibited hepatoprotective effects. Western blot analysis revealed that PQQ substantially downregulated the expressions of NLRP3 inflammasome signaling pathway proteins including NLRP3, thioredoxin interacting protein (TXNIP), apoptosis- associated speck - like protein containing a CARD (ASC), cysteinyl aspartate specific proteinase-1 (caspase-1), caspase-1 cut as well as IL-1β and interleukin-18 (IL-18). Conclusion PQQ significantly protects against diabetic liver damages in mice by enhancing antioxidant function and inhibiting NLRPS inflammasome signaling pathway.
关键词
吡咯喹啉醌 /
糖尿病 /
肝损伤 /
NLRP3炎症小体 /
小鼠
Key words
pyrroloquinoline quinone /
diabetes /
liver damage /
NLRP3 inflammasome /
mice
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] American Diabetes Association.Lifestyle management: standards of medical care in diabetes-2019[J]. Diabetes Care,2019,42:S46-S60.
[2] Regnell SE, Lernmark Å.Hepatic steatosis in type 1 diabetes[J]. Rev Diabet Stud, 2011,8:454–467.
[3] Thompson AJ, Patel K.Antifibrotic therapies: will we ever get there?[J]. Curr Gastroenterol Rep, 2010,12:23–29.
[4] Song JX, An JR, Chen Q, et al. Liraglutide attenuates hepatic iron levels and ferroptosis in db/db mice[J]. Bioengineered, 2022,13:8334–8348.
[5] Man SM, Karki R, Kanneganti TD.Molecular mechanisms and functions of pyroptosis, inflammatory caspases and inflammasomes in infectious diseases[J]. Immunol Rev, 2017,277:61–75.
[6] Wang S, Yuan YH, Chen NH, et al. The mechanisms of NLRP3 inflammasome/pyroptosis activation and their role in Parkinson's disease[J].Int Immunopharmaco, 2019,67:458-464.
[7] Martinon F, Burns K, Tschopp J.The inflammasome: a molecular platform triggering activation of inflammatory
caspases and processing of proIL-beta[J]. Mol Cell, 2002,10:417–426.
[8] Mridha AR, Wree A, Robertson AAB, et al. NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice[J]. J Hepatol, 2017,66:1037–1046.
[9] Kumar N, Kar A.Pyrroloquinoline quinone (PQQ) has potential to ameliorate streptozotocin-induced diabetes mellitus and oxidative stress in mice: a histopatho-logical and biochemical study[J]. Chem Biol Interact, 2015,240:278–90.
[10] Qu XF, Zhai BZ, Hu WL, et al. Pyrroloquinoline quinone ameliorates diabetic cardiomyopathy by inhibiting the pyroptosis signaling pathway in C57BL/6 mice and AC16 cells[J]. Eur J Nutr, 2022,61:1823–1836.
[11] Kumazawa T, Sato K, Seno H, et al. Levels of pyrrolo-quinoline quinone in various foods[J]. Biochem J, 1995,307:331–333.
[12] Kumazawa T, Seno H, Urakami T, et al. Trace levels of pyrroloquinoline quinone in human and rat samples detected by gas chromatography/mass spectrometry[J]. Biochim Biophys Acta, 1992,1156:62–66.
[13] Harris CB, Chowanadisai W, Mishchuk DO, et al. Dietary pyrroloquinoline quinone (PQQ) alters indicators of inflammation and mitochondrial-related metabolism in human subjects[J]. J Nutr Biochem, 2013,24:2076–2084.
[14] Wang Z, Li Y, Wang Y, et al. Pyrroloquinoline quinine protects HK-2 cells against high glucose-induced oxidative stress and apoptosis through Sirt3 and PI3K/Akt/FoxO3a signaling pathway[J]. Biochem Biophys Res Commun, 2019,508:398–404.
[15] Liu L, Zhang Y, Liu T, et al. Pyrroloquinoline quinone protects against exercise-induced fatigue and oxidative damage via improving mitochondrial function in mice[J]. FASEB J, 2021,35:e21394.
[16] Wen J, Shen J, Zhou Y, et al. Pyrroloquinoline quinone attenuates isoproterenol hydrochloride˗induced cardiac hypertrophy in AC16 cells by inhibiting the NF˗κB signaling pathway[J]. Int J Mol Med, 2020,45:873–885.
[17] Mridha AR, Wree A, Robertson AAB, et al. NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice[J]. J Hepatol, 2017,66:1037-1046.
[18] Bao X, Li J, Ren C, et al. Aucubin ameliorates liver fibrosis and hepatic stellate cells activation in diabetic mice via inhibiting ER stress-mediated IRE1α/TXNIP/NLRP3 inflammasome through NOX4/ROS pathway
[J]. Chem Biol Interact, 2022,365:110074.
[19] Wang A, Gong Y, Pei Z, et al Paeoniflorin ameliorates diabetic liver injury by targeting the TXNIP-mediated NLRP3 inflammasome in db/db mice[J]. Int Immunopharmacol, 2022,109:108792.
基金
大学生创新创业训练计划项目(No.202313023008); 浙江省医药卫生科技计划项目（No.2024KY924）
PDF(3858 KB)
