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International Journal of Genomics Volume 2017 ,2017-12-31
Lingguizhugan Decoction Protects against High-Fat-Diet-Induced Nonalcoholic Fatty Liver Disease by Alleviating Oxidative Stress and Activating Cholesterol Secretion
Research Article
Lili Yang 1 Weili Lin 2 , 3 Colleen A. Nugent 4 Shijun Hao 1 Haiyan Song 1 Tao Liu 1 Peiyong Zheng 1
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DOI:10.1155/2017/2790864
Received 2017-06-02, accepted for publication 2017-11-07, Published 2017-11-07
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摘要

Background. Nonalcoholic fatty liver disease (NAFLD) has become a leading cause of liver transplantation. Lingguizhugan decoction (LGZG), a classical Chinese herbal formula, has beneficial effects on NAFLD animal models. Our study examined the impact of LGZG on hepatic global transcriptome of high-fat-diet-induced NAFLD rats. Methods. Three groups of Wistar rats were included: normal, NAFLD model, and LGZG-treated NAFLD groups. Four weeks for the treatment, liver tissues were harvested for RNA sequencing. Differentially expressed genes (DEGs) and enriched pathways were detected on hepatic global transcriptome profile. Real-time PCR validated the regulatory patterns of LGZG on NAFLD rats. Results. DEGs between the NAFLD model and normal groups indicated the elevated peroxisome proliferator-activated receptor (PPAR) and hedgehog signaling pathways in NAFLD rats. In bile secretion pathway, genes involved in cholesterol secretion were activated by LGZG treatment. Increased expression of antioxidant OSIGN1 and decreased expression of genes (AHR, IRF2BP2, and RASGEF1B) that induce oxidative stress and inflammation were observed in NAFLD rats treated with LGZG. The regulatory patterns of LGZG treatment on these oxidative stress-related genes were confirmed by real-time PCR. Conclusion. Our study revealed a “two-hits-targeting” mechanism of LGZG in the treatment for NAFLD: alleviating oxidative stress and activating cholesterol secretion.

授权许可

Copyright © 2017 Lili Yang et al. 2017
This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

图表

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Median gene expression (a) was similar among all study groups. Housekeeping genes GAPDH (b), ACTB (c), G6PD (d), ALDOA (e), TAF (f), and GPI (g) and nonliver genes ATP4A (h), ATP4B (i), PGC (j), and MSN (k) exhibited similar expression among all study groups. ∗∗Q value < 0.01 calculated from cuffdiff.

Differential expression of top 40 NAFLD-regulated genes. The samples are broadly divided into two groups, normal and NAFLD model groups. The color scale shown at the top right illustrates the relative expression level of the genes across all samples. FC, fold change between NAFLD model and normal groups.

Scatter plot of the top 30 enriched KEGG pathways for NAFLD-regulated genes. The x-axis represents the ratio of NAFLD-regulated gene and all gene numbers annotated in this KEGG pathway. The y-axis is enriched KEGG pathways. P value was calculated from hypergeometric test. A smaller P value indicates higher significance (P<0.05).

Differential gene expression of (a) PPAR and (b) hedgehog signaling pathways in a NAFLD rat model. ∗Q value < 0.05; ∗∗Q value < 0.01; ∗∗∗Q value < 0.001; ∗∗∗∗Q value < 0.0001 calculated from cuffdiff. #Rate-limiting enzyme.

Differential gene expression of (a) PPAR and (b) hedgehog signaling pathways in a NAFLD rat model. ∗Q value < 0.05; ∗∗Q value < 0.01; ∗∗∗Q value < 0.001; ∗∗∗∗Q value < 0.0001 calculated from cuffdiff. #Rate-limiting enzyme.

Differential expression of top 40 LGZG-regulated genes. The samples of the NAFLD model group formed a clade and were separated from the samples of the LGZG-treated NAFLD groups. The color scale shown at the top right illustrates the relative expression level of the genes across all samples. FC, fold change between LGZG-treated NAFLD and NAFLD model groups.

Scatter plot of the enriched KEGG pathways for LGZG-regulated genes. The x-axis represents the ratio of LGZG-regulated gene numbers annotated in this KEGG pathway to all gene numbers annotated in this KEGG pathway. The y-axis is enriched KEGG pathways. P value was calculated from hypergeometric test. A smaller P value indicates higher significance (P<0.05).

Differential gene expression of LGZG-regulated genes. ∗Q value < 0.05; ∗∗Q value < 0.01 calculated from cuffdiff.

Differential gene expression of LGZG-regulated genes. ∗Q value < 0.05; ∗∗Q value < 0.01 calculated from cuffdiff.

Differential gene expression of LGZG-regulated genes. ∗Q value < 0.05; ∗∗Q value < 0.01 calculated from cuffdiff.

Differential gene expression of LGZG-regulated genes. ∗Q value < 0.05; ∗∗Q value < 0.01 calculated from cuffdiff.

Differential gene expression of LGZG-regulated genes was examined by real-time PCR. Plotted values are the mean ± SD of mRNA expression levels in the livers of normal rats (normal), NAFLD model rats, and NAFLD rats treated with LGZG decoction (LGZG). N=6 per group. ∗P<0.05; ∗∗P<0.01 Mann–Whitney test.

Differential gene expression of LGZG-regulated genes was examined by real-time PCR. Plotted values are the mean ± SD of mRNA expression levels in the livers of normal rats (normal), NAFLD model rats, and NAFLD rats treated with LGZG decoction (LGZG). N=6 per group. ∗P<0.05; ∗∗P<0.01 Mann–Whitney test.

Differential gene expression of LGZG-regulated genes was examined by real-time PCR. Plotted values are the mean ± SD of mRNA expression levels in the livers of normal rats (normal), NAFLD model rats, and NAFLD rats treated with LGZG decoction (LGZG). N=6 per group. ∗P<0.05; ∗∗P<0.01 Mann–Whitney test.

Differential gene expression of LGZG-regulated genes was examined by real-time PCR. Plotted values are the mean ± SD of mRNA expression levels in the livers of normal rats (normal), NAFLD model rats, and NAFLD rats treated with LGZG decoction (LGZG). N=6 per group. ∗P<0.05; ∗∗P<0.01 Mann–Whitney test.

Summary of the “two-hits-targeting” therapeutic mechanisms of LGZG for NAFLD. Data presented in this study supports two independent mechanisms: (i) LGZG activates antioxidant (OSGIN1) and suppresses prooxidant and proinflammatory genes (AHR, IRF2BP2, and RASGEF1B), causing alleviated oxidative stress, and (ii) LGZG induces cholesterol secretion genes in the liver, leading to reduced accumulation of liver lipid. These mechanisms reflect that LGZG treatment prominately impeded classic “two hits” of NASH physiopathology. Red genes indicate increased expression; green genes indicate decreased expression. Normal arrows indicate activation. Arrows with a vertical line at the end indicate suppression.

通讯作者

1. Tao Liu.Institute of Digestive Diseases, Longhua Hospital, China-Canada Center of Research for Digestive Diseases (ccCRDD), Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China, shutcm.edu.cn.lh2488@126.com
2. Peiyong Zheng.Institute of Digestive Diseases, Longhua Hospital, China-Canada Center of Research for Digestive Diseases (ccCRDD), Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China, shutcm.edu.cn.zpychina@sina.com

推荐引用方式

Lili Yang,Weili Lin,Colleen A. Nugent,Shijun Hao,Haiyan Song,Tao Liu,Peiyong Zheng. Lingguizhugan Decoction Protects against High-Fat-Diet-Induced Nonalcoholic Fatty Liver Disease by Alleviating Oxidative Stress and Activating Cholesterol Secretion. International Journal of Genomics ,Vol.2017(2017)

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