芪味铁屑胶囊通过调控 LXRα、PPARγ、NF-κB-iNOS-NO 信号通路对 3T3-L1 脂肪细胞及非酒精性脂肪性肝病大鼠的保护作用及机制研究。
Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic fatty liver disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.
机构信息
The Affliated Hospital of Tibet Tibetan Medical College, Tibet, Lhasa, 58000, China.
Department of General Surgey, The Orient Hospital of Beijing Traditional Medical University, Beijing, 100078, China.
出版信息
J Ethnopharmacol. 2019 May 23;236:316-325. doi: 10.1016/j.jep.2019.03.006. Epub 2019 Mar 6.
ETHNOPHARMACOLOGICAL RELEVANCE
Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD).
AIM OF THE STUDY
This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.
MATERIALS AND METHODS
The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat acid oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] palmitic acid, respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty acid (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.
RESULTS
QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.
CONCLUSIONS
QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.
民族药理学相关性
奇味铁屑胶囊（QWTX）是藏药的代表性处方，广泛用于慢性肝病和非酒精性脂肪肝（NAFLD）的长期治疗。
研究目的
本研究探讨了 QWTX 对 3T3-L1 脂肪细胞和 NAFLD 的作用和机制。
材料和方法
本研究使用了 3T3-L1 前脂肪细胞和 NAFLD 大鼠模型。在 3T3-L1 细胞中，通过 CKK-8 检测 QWTX 的细胞毒性，通过 2-脱氧-D-[H]葡萄糖和[1-C]棕榈酸分别评估葡萄糖摄取和脂肪酸氧化。通过 PCR 和 Western blot 测定肉毒碱棕榈酰转移酶-1（CPT-1）、肝 X 受体α（LXRα）、过氧化物酶体增殖物激活受体（PPAR）γ、诱导型一氧化氮合酶（iNOS）、Ikappa Bα（IκBα）和 AKT 的表达水平。通过给予脂肪乳液和蔗糖 9 周建立 NAFLD 模型。通过 HE 和透射电镜观察 QWTX 对 NAFLD 大鼠脂质代谢、肝功能和肝组织形态的影响。检测血清一氧化氮（NO）和游离脂肪酸（FFA）、超氧化物歧化酶（SOD）和丙二醛（MDA）含量以及细胞色素 P450 2E1（CYP2E1）、NF-κB、单核细胞趋化蛋白 1（MCP-1）、CPT-1、LXRα、PPARα、PPARβ/δ、PPARγ和 iNOS 的表达水平。
结果
QWTX 在 3T3-L1 前脂肪细胞中无细胞毒性，CO 生成率增加到 4.15，表明脂肪堆积减少。在 NAFLD 中，QWTX 减轻了肝脂肪变性、脂肪空泡和炎症，HE 染色和电镜检查结果均如此。对于氧化应激生物标志物，QWTX 治疗后血清 FFA 水平降低，血清 NO 水平升高。在肝组织中，NAFLD 时 SOD 降低，MDA 明显升高，QWTX 均可恢复。NF-κB 和 CYP2E1 在 NAFLD 中也上调，而 QWTX 则下调。QWTX 还观察到 3T3-L1 脂肪细胞和 NAFLD 模型中 LXRα、PPARγ 和 iNOS 的下调。
结论
QWTX 通过调节 LXRα、PPARγ 和 NF-κB-iNOS-NO 信号通路，保护分化的 3T3-L1 脂肪细胞和 NAFLD 中的肝损伤。
Zotero 插件