| 廖加抱,宋云,王斯,等.健肝消脂方调控PINK1/Parkin通路介导的线粒体自噬治疗非酒精性脂肪肝[J].浙江中医药大学学报,2024,48(8):905-914. |
| 健肝消脂方调控PINK1/Parkin通路介导的线粒体自噬治疗非酒精性脂肪肝 |
| Jian’gan Xiaozhi Decoction Regulates PINK1/Parkin Pathway Mediated Mitochondrial Autophagy to Treat Non-alcoholic Fatty Liver Disease |
| DOI：10.16466/j.issn1005-5509.2024.08.002 |
| 中文关键词: 健肝消脂方 非酒精性脂肪肝 PINK1/Parkin信号通路 线粒体自噬 氧化应激 炎症 |
| 英文关键词: Jiangan Xiaozhi Decoction non-alcoholic fatty liver disease PINK1/Parkin signaling pathway mitochondrial autophagy oxidative stress inflammation |
| 基金项目:国家自然科学基金青年项目(82104820)；浙江省自然科学基金项目(LTGY23H270009)；嘉兴市科技局应用性基础研究项目(2023AY11035) |
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| 中文摘要: |
| [目的] 探究健肝消脂方(Jian’gan Xiaozhi Decoction，JGXZ)对非酒精性脂肪肝(non-alcoholic fatty liver disease，NAFLD)模型小鼠的药效作用，并从人第10号染色体缺失的磷酸酶及张力蛋白同源基因诱导的激酶1(PTEN induced putative kinase 1，PINK1)/E3 泛素-蛋白连接酶(Parkin)信号通路介导的线粒体自噬角度探究其作用机制。[方法] 60只C57BL/6J小鼠适应性喂养1周后，随机分为6组，包括正常组(Control)，模型组(NAFLD)，多烯磷脂酰胆碱胶囊组(polyene phosphatidyl choline，PPC)和JGXZ低、中、高剂量组。除正常组外，另外5组均给予高脂饮食。治疗过程持续8周，每周统计小鼠体质量，8周后采集小鼠血液，分离血清，测定丙氨酸氨基转移酶(alanine aminotransferase，ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase，AST)水平；小鼠肝脏称重后固定，取同一部位进行苏木精-伊红(hematoxylin-eosin，HE)与油红O染色，观察肝组织病理学变化；采用试剂盒检测肝组织中的甘油三酯(triglyceride，TG)、总胆固醇(total cholesterol，TC)、白细胞介素-1β(interleukin-1β，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)、丙二醛(malondialdehyde，MDA)水平，以及谷胱甘肽过氧化物酶(glutathione peroxidase，GSH-Px)、超氧化物歧化酶(superoxide dismutase，SOD)活性，以评估JGXZ对NAFLD小鼠炎症反应及氧化应激的影响；免疫印迹法检测电压依赖性阴离子通道蛋白1(voltage dependent anion channel 1，VDAC1)、线粒体外膜转位酶20(translocase of outer mitochondrial membrane 20，TOM20)、细胞色素C氧化酶Ⅳ亚型(cytochrome c oxidase subunit Ⅳ，COXⅣ)、PINK1、Parkin、苄氯素1(Beclin1)、微管相关蛋白轻链3(light chain 3，LC3)以及选择性自噬接头蛋白 P62(prostacyclin 62，P62)蛋白表达，以评估JGXZ对NAFLD模型小鼠线粒体自噬的影响。[结果] 与模型组比较，JGXZ干预明显提升NAFLD模型小鼠体质量，降低肝指数，缓解NAFLD模型小鼠肝组织病变，降低TC、TG、ALT、AST水平，降低IL-1β、IL-6、TNF-α及MDA的水平，提高了GSH-Px、SOD活性，下调VDAC1、TOM20、COXⅣ以及P62蛋白表达，上调PINK1、Parkin、Beclin1、LC3蛋白表达。[结论] JGXZ可以通过促进PINK1/Parkin信号通路，介导线粒体自噬激活，缓解NAFLD小鼠肝损伤。 |
| 英文摘要: |
| [Objective] To investigate the therapeutic effect of Jian’gan Xiaozhi Decoction(JGXZ) on non-alcoholic fatty liver disease(NAFLD) model mice and explore its mechanism from the perspective of mitochondrial autophagy mediated by the PTEN induced putative kinase 1(PINK1)/E3 uniquitin-protein ligase(Parkin) signaling pathway. [Methods] Sixty C57BL/6J mice were randomly and equally divided into 6 ?groups: control， model(NAFLD)， polyene phosphatidyl choline(PPC) group 180 mg/kg intragastric administration， and JGXZ low， medium and high dose groups(8，16，32 ?g/kg) intragastric administration. Except for control group， the other 5 ?groups were given a high fat diet. The treatment lasted for 8 weeks， and the body weight of the mice was recorded weekly. After 8 weeks， blood samples were collected， the levels of serum alanine aminotransferase(ALT) and aspartate aminotransferase (AST) were measured， liver tissue was weighted and fixed， and histological changes in liver tissue were observed by hematoxylin-eosin(HE) and oil red O staining. The levels of total cholesterol(TC)， triglyceride(TG)， interleukin-1β(IL-1β)， IL-6， tumor necrosis factor-α(TNF-α)， malondialdehyde(MDA) and activities of glutathione peroxidase(GSH-Px)， superoxide dismutase(SOD) in liver tissue were measured to evaluate the effects of JGXZ on inflammation and oxidative stress in NAFLD mice. Western blot was used to detect the protein expression levels of voltage-dependent anion channel 1(VDAC1)， translocase of outer mitochondrial membrane 20(TOM20)， cytochrome c oxidase subunit Ⅳ(COXⅣ)， phosphatase and PINK1， Parkin， Beclin1， microtubule-associated protein light chain 3(LC3)， and P62 to evaluate the effects of JGXZ on mitochondrial autophagy in NAFLD mice. [Result] Compared with model group， JGXZ intervention significantly improved the body weight of NAFLD model mice， reduced liver index， alleviated liver tissue lesions in NAFLD model mice， reduced TC， TG， ALT， AST levels， decreased IL-1β， IL-6， TNF-α and MDA levels， increased GSH-Px and SOD activity， down-regulated VDAC1， TOM20， COXⅣ and P62 protein expression， and up-regulated PINK1， Parkin， Beclin1， LC3 protein expression. [Conclusion] JGXZ can alleviate liver injury in NAFLD mice by promoting the PINK1/Parkin signaling pathway mediated activation of mitochondrial autophagy. |
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