去脂软肝方对非酒精性脂肪性肝炎大鼠FXR-FGF19通路的影响

夏恩蕊; 田格格; 张素妍; 张顺贞

doi:10.3969/j.issn.1001-5256.2022.05.018

去脂软肝方对非酒精性脂肪性肝炎大鼠FXR-FGF19通路的影响

DOI: 10.3969/j.issn.1001-5256.2022.05.018

云南中医药大学中药学院，昆明 650000

基金项目:

国家自然科学基金 (81760818);

云南省高校中医证候微观辨证重点实验室 (20190720);

云南省科技厅中医联合专项 (2018FF001(-042));

云南省科技厅中医联合专项-重点项目 (202101AZ070001-008)

伦理学声明：本研究方案于2020年6月1日经由云南中医药大学实验动物伦理委员会审批，批号: R—06202016，符合实验室动物管理与使用准则。

利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。

作者贡献声明：夏恩蕊负责课题设计，资料分析，撰写论文；夏恩蕊、张素妍、田格格参与动物造模及实验，修改论文；张顺贞负责实验设计拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
张顺贞, 694415176@qq.com

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出版历程
- 收稿日期: 2021-09-27
- 录用日期: 2021-11-01
- 出版日期: 2022-05-20

Effect of Quzhi Ruangan prescription on the farnesoid X receptor-fibroblast growth factor 19 pathway in rats with nonalcoholic steatohepatitis

School of Traditional Chinese Medicine, Yunnan University of Traditional Chinese Medicine, Kunming 650000, China

Research funding:

The National Natural Science Foundation of China (81760818);

Key Laboratory of Microcosmic Syndrome Differentiation of TCM Syndrome in Yunnan Province (20190720);

Joint Special Project of Traditional Chinese Medicine of Yunnan Provincial Science and Technology Department (2018FF001(-042));

Joint Special Project of Traditional Chinese Medicine of Yunnan Provincial Science and Technology Department-Key Project (202101AZ070001-008)

More Information

Corresponding author: ZHAGN Shunzhen, 694415176@qq.com(ORCID: 0000-0003-3908-6414)

摘要

摘要: 目的观察去脂软肝方对非酒精性脂肪性肝炎(NASH)模型大鼠法尼醇X受体(FXR)-成纤维细胞因子19(FGF19)通路的影响。方法雄性SD大鼠随机分为正常组(Control组，n=8)、模型组(HFD组，n=12)、辛伐他汀组(Simvastatin组，n=8)、去脂软肝方高剂量组(QH组，n=8)、去脂软肝方低剂量组(QL组，n=8)，Control组给予普通饲料喂养，其余组高脂饲料喂养。10周末取材，观察大鼠肝脏病理切片变化，检测各组血清肝功能指标、肝脏与小肠FGF19、肝脏中胆汁酸(BA)。检测小肠FXR、肝脏胆固醇7α-羟化酶(CYP7A1)表达情况。多组间比较采用单因素方差分析，进一步两组间比较采用LSD-t检验。结果与Control组相比，HFD组病理表现出明显炎性病变和脂肪变性。与HFD组相比，各用药组的HDL-C显著升高，ALT、AST、TC、TG、LDL-C均显著下降(P值均＜0.05)。与Control组相比，HFD组大鼠小肠FGF19显著下降、肝脏BA显著升高(P值均＜0.05)。与HFD组相比，各用药组的小肠中FGF19显著升高，肝脏BA显著下降(P值均＜0.05)。与Control组相比，HFD组大鼠小肠FXR mRNA显著下降，肝脏CYP7A1 mRNA显著升高(P值均＜0.05)。与HFD组相比，QH组小肠FXR mRNA显著升高，QL组显著降低(P值均＜0.05)；QH组肝脏CYP7A1 mRNA显著下降(P＜0.05)。与Control组相比，HFD组大鼠小肠FXR阳性率显著下降，肝脏CYP7A1阳性率显著上升(P值均＜0.05)。与HFD组相比，Simvastatin组、QH组小肠FXR阳性率显著升高(P值均＜0.05)，Simvastatin组、QH组、QL组肝脏CYP7A1阳性率显著下降(P值均＜0.05)。结论去脂软肝方可激活NASH大鼠FXR-FGF19通路，并可能通过该途径对NASH产生防治作用。
- 非酒精性脂肪性肝炎 /
- 法尼醇X受体 /
- 细胞因子类
Abstract: Objective To investigate the effect of Quzhi Ruangan prescription on the farnesoid X receptor (FXR)-fibroblast growth factor 19 (FGF19) pathway in rats with nonalcoholic steatohepatitis (NASH). Methods Male Sprague-Dawley rats were randomly divided into normal group (Control group with 8 rats), model group (HFD group with 12 rats), simvastatin group with 8 rats, high-dose Quzhi Ruangan prescription group (QH group with 8 rats), and low-dose Quzhi Ruangan prescription group (QL group with 8 rats). The rats in the Control group were fed with a normal diet and those in the other groups were fed with a high-fat diet. Related samples were collected at the end of week 10 to observe liver pathological changes and measure the serum levels of liver function parameters, the level of FGF19 in the liver and the small intestine, and the level of bile acid (BA) in the liver. The expression levels of FXR in the small intestine and cholesterol 7α-hydroxylase (CYP7A1) in the liver were also measured. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison bewteen two groups. Results Compared with the Control group, the HFD group showed the pathological manifestations of marked inflammatory lesions and steatosis. Compared with the HFD group, all administration groups had a significant increase in high-density lipoprotein cholesterol and significant reductions in alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride, and low-density lipoprotein cholesterol (all P < 0.05). Compared with the Control group, the HFD group had a significant reduction in FGF19 in the small intestine and a significant increase in BA in the liver (both P < 0.05). Compared with the HFD group, all administration groups had a significant increase in FGF19 in the small intestine and a significant reduction in BA in the liver (all P < 0.05). Compared with the Control group, the HFD group had a significant reduction in the mRNA expression of FXR in the small intestine and a significant increase in the mRNA expression of CYP7A1 in the liver (both P < 0.05). Compared with the HFD group, the QH group had a significant increase in the mRNA expression of FXR in the small intestine, while the QL group had a significant reduction (both P < 0.05), and the QH group had a significant reduction in the mRNA expression of CYP7A1 in the liver (P < 0.05). Compared with the Control group, the HFD group had a significant reduction in the positive rate of FXR in the small intestine and a significant increase in the positive rate of CYP7A1 in the liver (both P < 0.05). Compared with the HFD group, the simvastatin group and the QH group had a significant increase in the positive rate of FXR in the small intestine (both P < 0.05), and the simvastatin group, the QH group, and the QL group had a significant reduction in the positive rate of CYP7A1 in the liver (all P < 0.05). Conclusion Quzhi Ruangan prescription can activate the FXR-FGF19 pathway in NASH rats and may exert a preventive and therapeutic effect on NASH through this pathway.
- Non-alcoholic Steatohepatitis /
- Farnesol X Receptor /
- Cytokines

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图 1 肝脏病理切片(×400)

Figure 1. Liver pathological section (×400)

下载: 全尺寸图片幻灯片

图 2 大鼠免疫组化切片(×400)

Figure 2. Immunohistochemical sections of rats(×400)

下载: 全尺寸图片幻灯片

表 1 大鼠肝脏油红O切片阳性率比较

Table 1. Comparison of positive rate of oil red O section of rat liver

组别	动物数(只)	油红O染色阳性率(%)
Control组	8	0.25±0.06
HFD组	8	20.50±2.18¹⁾
Simvastatin组	8	1.98±0.73²⁾
QH组	8	1.11±0.91²⁾
QL组	8	2.81±1.09¹⁾²⁾
F值		174.143
P值		＜0.05
注：与Control组相比，1)P＜0.05；与HFD组相比，2)P＜0.05。

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表 2 大鼠血脂、血清肝功能指标比较

Table 2. Comparison of blood lipids and serum liver function indexes in rats

组别	ALT (U/L)	AST (U/L)	TC (mmol/L)	TG (mmol/L)	HDL-C (mmol/L)	LDL-C (mmol/L)
Control组	34.62±3.11	125.37±25.23	1.32±0.11	0.41±0.06	0.87±0.06	0.22±0.04
HFD组	65.25±26.95¹⁾	229.50±80.13¹⁾	2.23±0.37¹⁾	0.81±0.14¹⁾	0.45±0.05¹⁾	1.29±0.35¹⁾
Simvastatin组	32.75±4.30²⁾	113.50±22.05²⁾	1.17±0.14¹⁾²⁾	0.32±0.07¹⁾²⁾	0.56±0.10¹⁾²⁾	0.48±0.07¹⁾²⁾
QH组	38.62±0.91²⁾	128.50±10.92²⁾	1.23±0.23²⁾	0.27±0.03¹⁾²⁾	0.62±0.17¹⁾²⁾	0.50±0.17¹⁾²⁾
QL组	35.62±2.77²⁾	148.00±13.68²⁾	1.22±0.18²⁾	0.28±0.06¹⁾²⁾	0.57±0.10¹⁾²⁾	0.61±0.11¹⁾²⁾
F值	9.57	11.11	30.98	56.01	17.45	37.17
P值	＜0.05	＜0.05	＜0.05	＜0.05	＜0.05	＜0.05
注：与Control组相比，1)P＜0.05；与HFD组相比，2)P＜0.05。

下载: 导出CSV

表 3 大鼠肝脏、小肠中FGF19和肝脏中BA水平比较

Table 3. Comparisons of FGF19 in liver, FGF19 in small intestine and BA in liver of rats

组别	小肠FGF19 (pg/mgpro)	肝脏FGF19 (pg/mgpro)	肝脏BA (nmol/mgpro)
Control组	597.49±38.61	92.33±32.25	3.43±0.51
HFD组	354.21±64.66¹⁾	77.52±14.99	7.54±0.30¹⁾
Simvastatin组	512.07±29.32¹⁾²⁾	116.33±18.06²⁾	4.31±0.30¹⁾²⁾
QH组	449.52±49.43¹⁾²⁾	180.06±217.92	5.26±0.60¹⁾²⁾
QL组	474.01±30.04¹⁾²⁾	86.16±12.05	5.73±0.45¹⁾²⁾
F值	31.98	1.40	94.35
P值	＜0.05	＜0.05	＜0.05
注：与Control组相比，1)P＜0.05；与HFD组相比，2)P＜0.05。

下载: 导出CSV

表 4 大鼠小肠FXR mRNA、肝脏CYP7A1 mRNA相对表达量比较

Table 4. FXR mRNA in the small intestine and CYP7A1 mRNA in the liver

组别	动物数 (只)	肝脏CYP7A1 mRNA	小肠FXR mRNA
Control组	6	0.30±0.09	0.90±0.10
HFD组	6	0.96±0.23¹⁾	0.33±0.04¹⁾
Simvastatin组	6	1.66±0.35¹⁾²⁾	0.34±0.07¹⁾
QH组	6	0.53±0.07¹⁾²⁾	1.08±0.25¹⁾²⁾
QL组	6	1.43±0.27¹⁾²⁾	0.01±0.00¹⁾²⁾
F值		71.23	36.94
P值		＜0.05	＜0.05
注：与Control组相比，1)P＜0.05；与HFD组相比，2)P＜0.05。

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表 5 小肠FXR、肝脏CYP7A1阳性率

Table 5. Comparisons of positive rates of FXR in small intestines and CYP7A1 in liver

组别	动物数 (只)	肝脏CYP7A1 阳性率(%)	小肠FXR 阳性率(%)
Control组	3	0.31±0.21	3.36±0.09
HFD组	3	1.56±1.18¹⁾	0.56±0.09¹⁾
Simvastatin组	3	0.09±0.14²⁾	3.34±0.09²⁾
QH组	3	0.42±0.44²⁾	2.84±0.09¹⁾²⁾
QL组	3	0.07±0.10²⁾	1.08±0.09¹⁾
F值		13.908	12.163
P值		＜0.05	＜0.05
注：与Control组相比，1)P＜0.05；与HFD组相比，2)P＜0.05。

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