Effect of total saponins from Panax japonicus on non-alcoholic steatohepatitis by regulating autophagy.
10.19540/j.cnki.cjcmm.20210201.401
- Author:
Yue LUO
1
;
Chao-Qi LIU
1
;
Hai-Bo HE
2
;
Ting WANG
3
;
Yu-Min HE
3
;
Chang-Cheng ZHANG
3
;
Ding YUAN
3
;
Cheng-Fu YUAN
3
Author Information
1. Hubei Key Laboratory of Tumor Microenvironment and Immunotheraphy, China Three Gorges University Yichang 443002, China.
2. College of Biology and Pharmaceuticals, China Three Gorges University Yichang 443002, China.
3. Medical College, China Three Gorges University Yichang 443002, China.
- Publication Type:Journal Article
- Keywords:
autophagy;
miR-199a-5p/ATG5;
non-alcoholic steatohepatitis;
total saponins of Panax japonicus
- MeSH:
Animals;
Autophagy;
Autophagy-Related Protein 5;
Mice;
MicroRNAs/genetics*;
Non-alcoholic Fatty Liver Disease/genetics*;
Panax;
Saponins/pharmacology*
- From:
China Journal of Chinese Materia Medica
2021;46(9):2260-2266
- CountryChina
- Language:Chinese
-
Abstract:
Non-alcoholic steatohepatitis(NASH) was induced by high-sugar and high-fat diet in mice to investigate the intervention effect of total saponins from Panax japonicus(TSPJ) and explore its possible mechanism. Mice were fed with high-sugar and high-fat diet to establish NASH model, and intervened with different doses of TSPJ(15, 45 mg·kg~(-1)). The animals were fed for 26 weeks. The histomorphology and pathological changes of liver tissues were observed by HE staining. The transcriptional expression levels of miR-199 a-5 p, autophagy related gene 5(ATG5) and inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) in mouse liver were measured by quantitative Real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the expression of autophagy-related proteins ATG5, P62/SQSTM1(P62), and microtubule-associated protein light chain 3(LC3)-I/Ⅱ proteins in mouse liver. The expression of P62 protein was detected by immunofluorescence staining. In order to verify the targeting regulation relationship between miR-199 a-5 p and ATG5, miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor were transfected into Hepa 1-6 cells, and the expression of ATG5 mRNA and protein was detected. pMIR-reportor ATG5-3'UTR luciferase reporter gene plasmid was constructed and co-transfected with miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor into Hepa 1-6 cells to detect luciferase activity. In vivo, HE staining in the model group showed typical fatty degeneration and inflammatory infiltration, with increased expression of miR-199 a-5 p and decreased expression of ATG5 mRNA and protein. The expression of autophagy-associated protein P62 increased significantly, the ratio of LC3Ⅱ/Ⅰ decreased, and the transcriptional expression of inflammatory factors increased significantly. After the intervention by TSPJ, the pathological performance of liver tissue was significantly improved, the expression of miR-199 a-5 p decreased and the expression of ATG5 mRNA and protein increased, the expression of autophagy-associated protein P62 decreased significantly, the ratio of LC3Ⅱ/Ⅰ increased, and the transcriptional expression of inflammatory cytokines IL-6, IL-1β and TNF-α decreased significantly. In vitro, it was found that the expression of ATG5 mRNA and protein and luciferase activity decreased significantly in miR-199 a-5 p overexpression cells, while after inhibition of miR-199 a-5 p expression, the expression level of ATG5 mRNA and protein and luciferase activity increased. The results showed that TSPJ can improve NASH in mice fed with high-sugar and high-fat diet, and its mechanism may be related to the regulation of miR-199 a-5 p/ATG5 signal pathway, the regulation of autophagy activity and the improvement of inflammatory response of NASH.
