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福建中医药大学,福州 350100
庄舒婷,博士,从事中西医结合防治糖尿病方向研究,E-mail:18344981481@163.com
* 施红,教授,从事中西医结合防治糖尿病方向研究,E-mail:shihong3327@sina.com
收稿日期:2020-05-13,
网络出版日期:2020-08-18,
纸质出版日期:2020-12-20
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庄舒婷,张家林,邹玉卿等.石斛合剂对2型糖尿病合并非酒精性脂肪肝大鼠AMPK/TFEB信号通路自噬蛋白的影响[J].中国实验方剂学杂志,2020,26(24):53-58.
ZHUANG Shu-ting,ZHANG Jia-lin,ZOU Yu-qing,et al.Effect of Shihu Mixture on Autophagy Protein of AMPK/TFEB Signaling Pathway in Rats with T2DM-NAFLD[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):53-58.
庄舒婷,张家林,邹玉卿等.石斛合剂对2型糖尿病合并非酒精性脂肪肝大鼠AMPK/TFEB信号通路自噬蛋白的影响[J].中国实验方剂学杂志,2020,26(24):53-58. DOI: 10.13422/j.cnki.syfjx.20202006.
ZHUANG Shu-ting,ZHANG Jia-lin,ZOU Yu-qing,et al.Effect of Shihu Mixture on Autophagy Protein of AMPK/TFEB Signaling Pathway in Rats with T2DM-NAFLD[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):53-58. DOI: 10.13422/j.cnki.syfjx.20202006.
目的
2
观察石斛合剂干预后,2型糖尿病(T2DM)合并非酒精性脂肪肝(NAFLD)大鼠腺苷酸活化蛋白激酶(AMPK)/转录因子EB(TFEB)自噬信号通路蛋白表达。
方法
2
40只雄性SD大鼠,根据体质量随机选择10只作为正常组。其余30只大鼠高脂高糖饮食喂养6周,然后腹腔注射链脲佐菌素建立2型糖尿病合并非酒精性脂肪肝模型。分为正常组(10 mL·kg
-1
·d
-1
),模型组(10 mL·kg
-1
·d
-1
),二甲双胍组(100 mg·kg
-1
·d
-1
),石斛合剂组(11.3 g·kg
-1
·d
-1
),每组大鼠灌胃4周。灌胃结束后,对大鼠实施安乐死。取腹主动脉血、肝组织,检测各组大鼠空腹血糖(FBG),血清中糖化血清蛋白(GSP),胰岛素(INS),甘油三酯(TG),总胆固醇(TC),高密度脂蛋白胆固醇(HDL-C),低密度脂蛋白胆固醇(LDL-C)含量;苏木素-伊红(HE)染色观察肝脏组织形态变化;蛋白免疫印迹法(Western blot)检测AMPK/TFEB信号通路相关蛋白的表达。
结果
2
与模型组比较,石斛合剂组与二甲双胍组大鼠的FBG,GSP下降(
P
<
0.05),INS升高(
P
<
0.05);与模型组比较,石斛合剂组与二甲双胍组大鼠HDL升高(
P
<
0.05),TC,TG,LDL含量明显降低(
P
<
0.05)。石斛合剂与二甲双胍均能一定程度上改善T2DM合并NAFLD大鼠肝脏形态。石斛合剂与二甲双胍p-AMPK/AMPK升高(
P
<
0.05),TFEB,LC3Ⅱ表达升高(
P
<
0.05)。
结论
2
石斛合剂可通过激活AMPK/TFEB自噬信号通路来改善糖脂代谢,改善肝脏病理形态。
Objective
2
To observe the expression of adenosine monophosphate activated protein kinase (AMPK)/transcription factor EB (TFEB) autophagy signaling pathway protein in type 2 diabetes (T2DM) complicated with nonalcoholic fatty liver disease (NAFLD) rats after intervention with Shihu mixture (SHM).
Method
2
Among 40 male SD rats, 10 rats were randomly selected as normal group according to body weight. The remaining 30 rats were fed with high-fat and high-sugar diet for 6 weeks, and then intraperitoneally injected with streptozotocin (STZ) to establish a T2DM NAFLD model. They were divided into normal control group (10 mL·kg
-1
·d
-1
), model control group (10 mL·kg
-1
·d
-1
), metformin group (100 mg·kg
-1
·d
-1
), SHM group (11.3 g·kg
-1
·d
-1
). The rats in each group were gavaged for 4 weeks. After gavage, the rats were euthanized. Abdominal aortic blood and liver tissue were collected to detect fasting blood glucose (FBS), glycated serum protein (GSP), insulin (INS), triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C) content. Htoxylin eosin (HE) staining was performed to observe changes in liver tissue morphology. Western blot was used to detect AMPK/TFEB signaling pathway-related protein expression.
Result
2
Compared with the model control group, FBS and GSP of the SHM group and the DMBG group decreased (
P
<
0.05), while INS increased (
P
<
0.05). Compared with the model group, HDL increased in the SHM group and the DMBG group (
P
<
0.05), whereas TC, TG and LDL contents decreased (
P
<
0.05). Liver HE staining results showed that both SHM and Metformin could improve the liver morphology of T2DM and NAFLD rats to some extent. Western blot results showed that p-AMPK/AMPK of SHM and metformin increased (
P
<
0.05), while the expressions of TFEB and LC3Ⅱ increased (
P
<
0.05).
Conclusion
2
SHM can improve glucose and lipid metabolism by activating AMPK/TFEB autophagy signaling pathway, so as to improve liver pathological morphology.
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