| 李丽,廖广辉,楼招欢,等.竹节参醇提物对小鼠酒精性脂肪肝的影响[J].浙江中医药大学学报,2022,46(12):1310-1319. |
| 竹节参醇提物对小鼠酒精性脂肪肝的影响 |
| Effect of Ethanol Extract of Rhizoma Panax Japonicus on Alcoholic Fatty Liver in Mice |
| DOI：10.16466/j.issn1005-5509.2022.12.002 |
| 中文关键词: 浙派本草 本草纲目拾遗 酒精性脂肪肝 竹节参 胆固醇 氧化应激 |
| 英文关键词: Zhejiang school materia medica Collection of Compendium of Materia Medica alcoholic fatty liver Rhizoma Panax japonicas cholesterol oxidative stress |
| 基金项目:浙江省中医药科技计划(2019ZA041)；杭州市科技计划引导项目(20191231Y201) |
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| 中文摘要: |
| [目的] 观察竹节参醇提物对急性酒精性脂肪肝的保护作用并初步探讨其作用机制。[方法] 将48只ICR小鼠随机分为正常对照组、模型组、阳性对照组(多烯磷脂酰胆碱胶囊 0.27 g·kg-1)、竹节参低剂量组(0.1 g·kg-1)、竹节参中剂量组(0.2 g·kg-1)、竹节参高剂量组(0.4 g·kg-1)。小鼠高脂饲料喂养，以红星二锅头酒梯度稀释液为造模试剂，连续灌胃8周，复制酒精性脂肪肝模型，同时各给药组以相应药物进行干预。以全自动生化仪检测血清总胆固醇(total cholesterol，TC)、甘油三酯(triglyceride，TG)、谷丙转氨酶(alanine transaminase，ALT)、谷草转氨酶(aspertate aminotransferase，AST)水平；苏木精-伊红(hematoxylin-eosin，HE)染色观察肝组织损伤程度；油红O染色法观察肝脏脂肪沉积情况；以试剂盒测定肝组织匀浆超氧化物歧化酶(superoxide dismutase，SOD)活性及丙二醛(malondialdehyde，MDA)水平；免疫印迹法测定小鼠肝脏胰岛素诱导基因1编码蛋白(insulin induced gene 1，Insig1)、低密度脂蛋白受体(low-density lipoprotein receptor，LDLR)、过氧化物酶体增殖物激活受体α(peroxisome proliferator-activated receptor α， PPARα)、胆固醇调节元件结合蛋白1(sterol-regulatory element binding proteins 1，SREBP1)、甾醇调节因子结合蛋白裂解激活蛋白(sterol cleavage-activating protein，SCAP)、磷酸化氨基末端蛋白激酶(phosphorylated amino terminal protein kinase，p-JNK)蛋白表达；实时荧光定量聚合酶链式反应(Real-time fluorescence quantitative polymerase chain reaction，Real-time qPCR)检测肝脏Insig1、LDLR、PPARα、SREBP1、SCAP基因表达。[结果] 竹节参醇提物能下调小鼠体质量及肝脏指数，明显改善小鼠血清ALT、AST水平(P＜0.05，P＜0.01)，各剂量组间TC、TG水平差异无统计学意义(P＞0.05)。HE染色显示，竹节参中、高剂量组肝细胞饱满，无炎性浸润细胞；油红O染色显示，竹节参中、高剂量组能明显改善小鼠肝脏脂肪累积。竹节参中剂量组明显提升肝脏SOD活性(P＜0.05)，低、高剂量组能明显降低肝脏MDA水平(P＜0.05)。竹节参中、高剂量组能够显著降低Insig1、SCAP蛋白表达(P＜0.01)，各剂量组均能显著降低SREBP1、P-JNK/JNK蛋白相对表达水平(P＜0.01)，显著提升LDLR、PPARα蛋白相对表达水平(P＜0.05，P＜0.01)。此外，竹节参中、高剂量组还能显著降低Insig1、SCAP、SREBP1的mRNA 基因相对表达水平(P＜0.05，P＜0.01)，显著提升LDLR、PPARα mRNA基因相对表达水平(P＜0.05，P＜0.01)。 [结论] 竹节参醇提物对酒精性脂肪肝具有保护作用，该作用可能与改善肝脏胆固醇转运及合成，抑制脂质过氧化有关。 |
| 英文摘要: |
| [Objective] To study the protective effect and the mechanism of action of ethanol extract of Rhizoma Panax japonicus on acute alcoholic fatty liver(AFL). [Methods] Forty-eight mice were randomly divided into normal control group， model group， positive control group(Polyene phosphatidylcholine capsule 0.27 g·kg-1)， ethanol extract of Rhizoma Panax japonicus low dose group(0.1 g·kg-1)， ethanol extract of Rhizoma Panax japonicus middle dose group(0.2 g·kg-1) and ethanol extract of Rhizoma Panax japonicus high dose group(0.4 g·kg-1). AFL mice model of was established by intragastric administration of Hongxing Chinese Vodka for 8 weeks. After final experiment， serum total cholesterol(TC)， triglyceride(TG)， alanine transaminase(ALT)， aspertate aminotransferase(AST) levels were detected by fully automatic biochemical instrument； hematoxylin-eosin(HE) staining was used to observe pathological changes； oil red O staining was used to examine hepatic lipidosis； the activity of superoxide dismutase(SOD) and level of malondialdehyde(MDA) in liver tissue homogenate were determined with kit； the expressions of insulin induced gene 1(Insig1)， low-density lipoprotein receptor(LDLR)， peroxisome proliferator-activated receptor α(PPARα)， sterol-regulatory element binding proteins 1(SREBP1)， sterol cleavage-activating protein(SCAP) and phosphorylated amino terminal protein kinase(p-JNK) proteins were determined by Western blot； the expression levels of Insig1， LDLR， SCAP， SREBP1 and PPARα were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time qPCR). [Results] Ethanol extract of Rhizoma Panax japonicus high dose group could reduce the body weight and liver index of AFL mice， obviously improve the mice serum ALT， AST levels(P＜0.05， P＜0.01)， but there was no statistical difference of serum TC， TG levels among the ethanol extract of Rhizoma Panax japonicus groups(P＞0.05). HE staining showed that the liver cells in ethanol extract of Rhizoma Panax japonicus middle and high dose groups were full， without inflammatory infiltrating cells； oil red O staining showed that ethanol extract of Rhizoma Panax japonicus middle and high dose groups significantly reduced the fat accumulation in the liver cell. The SOD activity was significantly elevated in ethanol extract of Rhizoma Panax japonicus middle dose group(P＜0.05)， and the MDA level was dropped significantly in ethanol extract of Rhizoma Panax japonicus low and high dose groups(P＜0.05). Meanwhile， middle and high dose ethanol extract of Rhizoma Panax japonicus obviously decreased the protein expression levels of Insig1 and SCAP(P＜0.01)， the ethanol extract of Rhizoma Panax japonicus groups could significantly decrease SREBP1 and p-JNK/JNK protein expression(P＜0.01)， and obviously increased the protein expression of LDLR and PPARα(P＜0.05， P＜0.01). In addition， ethanol extract of Rhizoma Panax japonicus middle and high dose groups could effectively reduce the relative mRNA expression of Insig1， SCAP and SREBP1(P＜0.05， P＜0.01)， and increase clearly the relative mRNA expression of LDLR and PPARα(P＜0.05， P＜0.01). [Conclusion] Ethanol extract of Rhizoma Panax japonicus can significantly improve the transport and synthesis of cholesterol， and the effect is associated with inhibition of lipid peroxidation. |
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