1.湖北中医药大学针灸骨伤学院,湖北省武汉市武昌区昙华林特1号,430061
2.江汉大学医学部
pxlzhenjiu@163.com
收稿:2023-07-28,
修回:2023-09-11,
纸质出版:2023-11-17
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金舒文,刘嘉宝,李丹等.电针“丰隆”穴对高脂血症模型大鼠肝脏脂质合成和胰岛素抵抗的影响[J].中医杂志,2023,64(22):2346-2353.
JIN Shuwen,LIU Jiabao,LI Dan,et al.Effect of Electroacupuncture at “Fenglong”(ST 40) on Liver Lipid Synthesis and Insulin Resistance in Hyperlipidemic Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(22):2346-2353.
金舒文,刘嘉宝,李丹等.电针“丰隆”穴对高脂血症模型大鼠肝脏脂质合成和胰岛素抵抗的影响[J].中医杂志,2023,64(22):2346-2353. DOI: 10.13288/j.11-2166/r.2023.22.014.
JIN Shuwen,LIU Jiabao,LI Dan,et al.Effect of Electroacupuncture at “Fenglong”(ST 40) on Liver Lipid Synthesis and Insulin Resistance in Hyperlipidemic Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(22):2346-2353. DOI: 10.13288/j.11-2166/r.2023.22.014.
目的
2
从肝脏脂质合成和胰岛素抵抗探讨电针丰隆穴治疗高脂血症的可能机制。
方法
2
将18只大鼠随机分为空白组、模型组、电针组,每组6只。模型组和电针组大鼠予以高脂饲料喂养8周制备高脂血症模型。造模后电针组大鼠进行双侧“丰隆”穴电针治疗,模型组大鼠每日捆绑处理,每天1次,每周连续5天,共4周。空白组不予以特殊处理。干预后采用ELISA法检测各组大鼠肝组织甘油三酯(TG)、游离脂肪酸(FFA)含量;检测空腹血糖(FBG)及血清TG、FFA、空腹胰岛素(FINS)、丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、肿瘤坏死因子-ɑ(TNF-ɑ)、白细胞介素6(IL-6)含量,计算胰岛素抵抗指数(HOMA-IR)。HE染色和油红O染色观察肝脏病理情况。免疫荧光法检测肝脏固醇调节元结合蛋白1c(SREBP1c)、脂肪酸合酶(FASN)和硬脂酰辅酶A去饱和酶1(SCD1)的表达情况。Western blot法检测肝脏胰岛素受体底物1(IRS1)和酪氨酸磷酸化胰岛素受体底物1(p-IRS1-Tyr)蛋白表达水平。
结果
2
与空白组比较,模型组大鼠血清和肝组织TG、FFA含量,血清AST、ALT和TNF-ɑ、IL-6含量,FBG、FINS和HOMA-IR升高(
P
<0.05);HE染色可见肝细胞排列紊乱,呈广泛脂肪样变,油红O染色可见肝细胞质内大量鲜红色脂滴,脂质蓄积严重;肝组织SREBP1c、FASN、SCD1平均荧光强度显著上升(
P
<0.05);p-IRS1-Tyr蛋白表达水平显著降低(
P
<0.05)。与模型组比较,电针组大鼠血清和肝组织TG、FFA含量显著降低,血清AST、ALT和TNF-ɑ、IL-6含量明显下降,FBG、FINS和HOMA-IR降低(
P
<0.05);HE 染色可见肝细胞排列较规则,油红O染色可见肝细胞脂滴显著减少,脂质蓄积程度减轻;肝组织SREBP1c、FASN、SCD1平均荧光强度显著下降(
P
<0.05);p-IRS1-Tyr蛋白表达水平均显著升高(
P
<0.05)。
结论
2
电针“丰隆”穴可显著降低高脂血症模型大鼠血脂水平,改善肝脏脂肪堆积,提高胰岛素敏感性,其机制可能与抑制相关肝脏脂质合成分子表达、降低血清炎症因子、升高胰岛素底物受体磷酸化水平有关。
Objective
2
To explore the effect and possible mechanism of electroacupuncture (EA) at Fenglong (ST40) on liver lipid synthesis and insulin resistance (IR) in hyperlipidemic (HLP) rats.
Method
2
Eighteen rats were randomly divided into three groups, blank group, model group, and EA group, each consisting of six rats. The blank group rats were with fed a basic diet, while those in the model group and EA group were fed high-fat diet for 8 weeks. After modeling, the rats in the EA group received bilateral EA treatment at “Fenglong” (ST 40). The rats in the model group underwent daily binding treatment, once a day, continuously 5 days a week, for a total of 4 weeks. Following the intervention, the levels of triglycerides (TG) and free fatty acids (FFA) in liver tissue was determined using ELISA. Serum TG, FFA, fasting insulin (FINS), alanine transaminase (ALT), aspartate Transaminase (AST), tumor necrosis factor-ɑ (TNF-ɑ)and interleukin 6 (IL-6) were also measured. The fasting plasma glucose (FBG) assessed using a glucose meter and the homeostatic model assessment of insulin resistance (HOMA-IR) was calculated. Liver pathology was examined through HE staining and oil red O staining. The expression of hepatic sterol regulator binding protein 1c (SREBP1c), recombinant fatty acid synthase (FASN) and stearoyl-CoA desaturase 1 (SCD1) were detected through immunofluorescence. The protein expression levels of liver insulin receptor substrate 1 (IRS1) and tyrosine-phosphorylated insulin receptor substrate 1 (p-IRS1-Tyr) were determined via Western blot.
Results
2
When compared to the blank group, the model group of rats exhibited elevated serum and liver tissue levels of TG and FFA, as well as increased serum levels of AST, ALT, TNF-α, IL-6, FBG, FINS, and HOMA-IR (
P
<0.05). HE staining revealed disordered arrangements of liver cells, indicating widespread fatty degeneration. Oil red O staining showed abundant bright red lipid droplets within liver cell cytoplasm, indicating severe lipid accumulation. The average fluorescence intensity of SREBP1c, FASN, and SCD1 in liver tissue significantly increased (
P
<0.05), while p-IRS1-Tyr protein expression levels significantly decreased (
P
<0.05). In comparison to the model group, the EA group of rats showed significantly reduced serum and liver tissue levels of TG and FFA, along with decreased serum levels of AST, ALT, TNF-α, IL-6, FBG, FINS, and HOMA-IR (
P
<0.05). HE staining indicated more regular arrangements of liver cells, and oil red O staining revealed a significant reduction in liver cell lipid droplets, indicating a less severe degree of lipid accumulation. The average fluorescence intensity of SREBP1c, FASN, and SCD1 in liver tissue significantly decreased (
P
<0.05), while p-IRS1-Tyr protein expression levels significantly increased (
P
<0.05), with no significant difference in IRS1 protein expression (
P
>0.05).
Conclusion
2
EA at “Fenglong” (ST 40) can significantly decrease serum lipid in HLP rats, improves liver fat accumulation, and also ameliorate insulin resistance. The mechanism may be related to the inhibition of hepatic lipid synthesis molecule expression, reduced serum inflammatory factors, and an increase in insulin substrate receptor phosphorylation levels.
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