Effects of Tanshinone IIA on the modulation of miR‑33a and the SREBP‑2/Pcsk9 signaling pathway in hyperlipidemic rats

Jia,Lianqun; Song,Nan; Yang,Guanlin; Ma,Yixin; Li,Xuetao; Lu,Ren; Cao,Huimin; Zhang,Ni; Zhu,Meilin; Wang,Junyan; Leng,Xue; Cao,Yuan; Du,Ying; Xu,Yue

doi:10.3892/mmr.2016.5133

Effects of Tanshinone IIA on the modulation of miR‑33a and the SREBP‑2/Pcsk9 signaling pathway in hyperlipidemic rats

Authors:
- Lianqun Jia
- Nan Song
- Guanlin Yang
- Yixin Ma
- Xuetao Li
- Ren Lu
- Huimin Cao
- Ni Zhang
- Meilin Zhu
- Junyan Wang
- Xue Leng
- Yuan Cao
- Ying Du
- Yue Xu
View Affiliations

Affiliations: Key Laboratory of Ministry of Education for Traditional Chinese Medicine Viscera‑State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China, Graduate School, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China, The First Clinical College, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China
Published online on: April 14, 2016 https://doi.org/10.3892/mmr.2016.5133
Pages: 4627-4635
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Tanshinone IIA is the active compound isolated from Salvia miltiorrhiza bunge, which is a traditional Chinese medicine known as Danshen. The aim of the present study was to assess the effect of Tanshinone IIA on the regulation of lipid metabolism in the livers of hyperlipidemic rats and the underlying molecular events. An in vivo model of hyperlipidemia was established in rats, with the animals receiving a daily dose of Tanshinone IIA. The serum lipid profiles were analyzed using an automatic biochemical analyzer, and the histopathological alterations and lipid deposition in liver tissue were assessed using hematoxylin and eosin staining, and oil red O staining, respectively. The mRNA expression levels of microRNA (miR)‑33a, ATP‑binding cassette transporter (ABC)A1, ABCG1, sterol regulatory element‑binding protein 2 (SREBP‑2), proprotein convertase subtilisin/kexin type 9 (Pcsk9) and low‑density lipoprotein receptor (LDL‑R) in liver tissues were measured using reverse transcription‑quantitative polymerase chain reaction, and the protein expression levels of ABCA1, ABCG1, SREBP‑2, Pcsk9, and LDL‑R were analyzed using western blotting. Tanshinone IIA reduced lipid deposition and improved histopathology in the rat liver tissue, however, did not alter the lipid profile in rat serum. In addition, Tanshinone IIA treatment suppressed the expression of miR‑33a, whereas the protein expression levels of ABCA1, SREBP‑2, Pcsk9 in addition to LDL‑R mRNA and protein were upregulated. In conclusion, the present study indicated that Tanshinone IIA attenuated lipid deposition in the livers of hyperlipidemic rats and modulated the expression of miR‑33a and SREBP‑2/Pcsk9 signaling pathway proteins.

View Figures

View References

1	Chait A and Brunzell JD: Acquired hyperlipidemia (secondary dyslipoproteinemias). Endocrinol Metab Clin North Am. 19:259–278. 1990.PubMed/NCBI
2	Williams AD: Hyperlipidaemia and atherogenesis. Med Hypotheses. 33:213–217. 1990. View Article : Google Scholar : PubMed/NCBI
3	Xie Y, He YB, Zhang SX, Pan AQ, Zhang J, Guan XH, Wang JX and Guo WS: Treatment of combined hyperlipidemia patients by jiangzhi tongluo soft capsule combined atorvastatin calcium tablet: A clinical study. Chin J Int Trad Western Med. 34:1059–1063. 2014.In Chinese.
4	Fruebis J, Bird DA, Pattison J and Palinski W: Extent of antioxidant protection of plasma LDL is not a predictor of the antiatherogenic effect of antioxidants. J Lipid Res. 38:2455–2464. 1997.
5	Tall AR, Yvan-Charvet L, Terasaaka N, Pagler T and Wang N: HDL, ABC transporters and cholesterol efflux: Implications for the treatment of atherosclerosis. Cell Metab. 7:365–375. 2008. View Article : Google Scholar : PubMed/NCBI
6	Tang F, Wu X, Wang T, Wang P, Li R, Zhang H, Gao J, Chen S, Bao L, Huang H and Liu P: Tanshinone IIA attenuates atherosclerotic calcification in rat model by inhibition of oxidative stress. Vascul Pharmacol. 46:427–438. 2007. View Article : Google Scholar : PubMed/NCBI
7	Gao S, Liu Z, Li H, Little PJ, Liu P and Xu S: Cardiovascular actions and therapeutic potential of Tanshinone IIA. Atherosclerosis. 220:3–10. 2012. View Article : Google Scholar
8	Xu W, Yang J and Wu LM: Cardioprotective effects of anshinone IIA on myocardial ischemia injury in rats. Pharmazie. 64:332–336. 2009.PubMed/NCBI
9	Gong Z, Huang C, Sheng X, Zhang Y, Li Q, Wang MW, Peng L and Zang YQ: The role of Tanshinone IIA in the treatment of obesity through peroxisome proliferator-activated receptor gamma antag-onism. Endocrinology. 150:104–113. 2009. View Article : Google Scholar
10	Tang FT, Cao Y, Wang TQ, Wang LJ, Guo J, Zhou XS, Xu SW, Liu WH, Liu PQ and Huang HQ: Tanshinone IIA attenuates atherosclerosis in ApoE (−/−) mice through down-regulation of scavenger receptor expression. Eur J Pharmacol. 650:275–884. 2011. View Article : Google Scholar
11	Jia LQ, Feng JY, Yang GL, Chen WN and Chen Y: Effect of Tanshinone IIA on TLR4 and TNF-α of endothelial cells induced by LPS. Chin J Cell Mol Immunol. 27:733–735. 2011.In Chinese.
12	Jia LQ, Yang GL, Ren L, Chen WN, Feng JY, Gao Y, Zhang L, Li XT and Lei P: Tanshinone IIA reduces apoptosis induced by hydrogen peroxide in the human endothelium-derived EA.hy926 cells. J Ethnopharmacol. 143:100–108. 2012. View Article : Google Scholar : PubMed/NCBI
13	Horie T, Ono K, Horiguchi M, Nishi H, Nakamura T, Nagao K, Kinoshita M, Kuwabara Y, Marusawa H, Iwanaga Y, et al: MicroRNA-33 encoded by an intron of sterol regulatory element-binding protein 2 (SREBP-2l) regulates HDL in vivo. Proc Natl Acad Sci USA. 107:17321–17326. 2010. View Article : Google Scholar
14	Dávalos A, Goedeke L, Smibert P, Ramirez CM, Warrier NP, Andreo U, Cirera-Salinas D, Rayner K, Suresh U, Pastor-Pareja JC, et al: miR-33a/b contribute to the regulation of fatty acid metabolism and insulin signaling. Proc Natl Acad Sci USA. 108:9232–9237. 2011. View Article : Google Scholar : PubMed/NCBI
15	Wu CY, Tang ZH, Liu LS and Jiang ZS: Selecting pharmacological targets of Pcsk9. Chin J Biochem Mol Biol. 25:991–996. 2009.In Chinese.
16	Costet P, Cariou B, Lambert G, Lalanne F, Lardeux B, Jarnoux AL, Grefhorst A, Staels B and Krempf M: Hepatic Pcsk9 expression is regulated by nutritional status via insulin and sterol regulatory element-binding protein 1c. J Biol Chem. 281:6211–6218. 2006. View Article : Google Scholar : PubMed/NCBI
17	Attie AD: The mystery of Pcsk9. Arterioscler Thromb Vasc Biol. 24:1337–1339. 2004. View Article : Google Scholar : PubMed/NCBI
18	Steinberg D and Witztum JL: Inhibition of Pcsk9: A powerful weapon for achieving ideal LDL cholesterol levels. Proc Natl Acad Sci USA. 106:9546–9547. 2009. View Article : Google Scholar : PubMed/NCBI
19	Dong B, Wu MH, Li H, Kraemer FB, Adeli K, Seidah NG, Park SW and Liu J: Strong iduction of Pcsk9 gene expression through HNF1alpha and SREBP-2: Mechanism for the resistance to LDL-cholesterol lowering effect of statins in dyslipidemic hamsters. J Lipid Res. 51:1486–1495. 2010. View Article : Google Scholar : PubMed/NCBI
20	Najafi-Shoushtari SH, Kristo F, Li Y, Shioda T, Cohen DE, Gerszten RE and Näär AM: MicroRNA-33 and the SREBP host genes cooperate to control cholesterol homeostasi. Science. 328:1566–1569. 2010. View Article : Google Scholar : PubMed/NCBI
21	Attie AD: ABCA1: At the nexus of cholesterol, HDL and atherosclerosis. Trends Biochem Sci. 32:172–179. 2007. View Article : Google Scholar : PubMed/NCBI
22	Fernández-Hernando C, Ramírez CM, Goedeke L and Suárez Y: MicroRNAs in metabolic disease. Arterioscler Thromb Vasc Bio. 33:178–185. 2013. View Article : Google Scholar
23	Gerin I, Clerbaux LA, Haumont O, Lanthier N, Das AK, Burant CF, Leclercq IA, MacDougald OA and Bommer GT: Expression of miR-33 froman SREBP-2 intron inhibits cholesterol export and fatty acid oxidation. J Biol Chem. 285:33652–33661. 2010. View Article : Google Scholar : PubMed/NCBI
24	Fernández-Hernando C, Suárez Y, Rayner KJ and Moore KJ: MicroRNAs in lipid metabolism. Curr Opin Lipidol. 22:86–92. 2011. View Article : Google Scholar :
25	Lewis GF and Rader DJ: New insights into the regulation of HDL metabolism and reverse cholesterol transport. Circ Res. 96:1221–1232. 2005. View Article : Google Scholar : PubMed/NCBI
26	Rayner KJ, Sheedy FJ, Esau CC, Hussain FN, Temel RE, Parathath S, van Gils JM, Rayner AJ, Chang AN, Suarez Y, et al: Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. J Clin Invest. 121:2921–2931. 2011. View Article : Google Scholar : PubMed/NCBI
27	Rayner KJ, Esau CC, Hussain FN, McDaniel AL, Marshall SM, van Gils JM, Ray TD, Sheedy FJ, Goedeke L, Liu X, et al: Inhibition of miR-33a/b in non-human primates raises plasma HDL and lowera VLDL triglycerides. Nature. 478:404–447. 2011. View Article : Google Scholar : PubMed/NCBI
28	Liu W, Zhai C, Zhang X, Zhang H, Jiang M, Zhou S, Liu Y, Zhao N and Zhao J: Research of the whole grain-soybean compound package to regulate the cholesterol metabolism by SREBP-2, LDLR and visfatin. J Hyg Res. 42:196–202. 2013.In Chinese.
29	Xu W, Liu L and Homby D: c-IAP1 binds and processes Pcsk9 protein: Linking the c-IAP1 in a TNF-α pathway to Pcsk9-mediated LDLR degradation pathway. Molecules. 17:12086–12101. 2012. View Article : Google Scholar : PubMed/NCBI
30	Mani DN, Bawankule D and Saroj BK: Hyperlipidemic model: Studying lipid profile in small experimental animal. Int J Pharmacy Pharm Sci. 4:337–340. 2012.
31	Munshi RP, Joshi SG and Rane BN: Development of an experimental diet model in rats to study hyperlipidemia and insulin resistance, markers for coronary heart disease. Indian J Pharmacol. 46:270–246. 2014. View Article : Google Scholar : PubMed/NCBI
32	Omagari K, Kadokawa Y, Masuda J, Egawa I, Sawa T, Hazama H, Ohba K, Isomoto H, Mizuta Y, Hayashida K, et al: Fatty liver in non-alcoholic non overweight Japanese adults: Incidence and clinical characteristics. J Gastroenterol Hepatol. 17:1098–1105. 2002. View Article : Google Scholar : PubMed/NCBI
33	Hilden M, Christoffersen P, Juhl E and Dalgaard JB: Liver histology in a 'normal' population-examinations of 503 consecutive fatal traffic casualties. Scand J Gastroenterol. 12:593–597. 1997. View Article : Google Scholar
34	Shen L, Fan JG, Shao Y, Zeng MD, Wang JR, Luo GH, Li JQ and Chen SY: Prevalence of nonalcoholic fatty liver among administrative officers in Shanghai: An epidemiological survey. World J Gastroenterol. 9:1106–1110. 2003. View Article : Google Scholar : PubMed/NCBI
35	Ikonen E: Cellular cholesterol trafficking and compartmentalization. Nat Rev Mol Cell Biol. 9:125–138. 2008. View Article : Google Scholar : PubMed/NCBI
36	Sone H, Tanaka S, Tanaka S, Iimuro S, Oida K, Yamasaki Y, Oikawa S, Ishibashi S, Katayama S, Ohashi Y, et al: Serum level of triglycerides is a potent risk factor comparable to LDL cholesterol for coronary heart disease in Japanese patients with type 2 diabetes: Subanalysis of the Japan Diabetes Complications Study (JDCS). J Clin Endocrinol Metab. 96:3448–3456. 2011. View Article : Google Scholar : PubMed/NCBI
37	Brown MS and Goldstein JL: A receptor-mediated pathway for cholesterol homeostasis. Science. 232:34–47. 1986. View Article : Google Scholar : PubMed/NCBI
38	Sharpe LJ and Brown AJ: Rapamycin down-regulates LDL-receptor expression independently of SREBP-2. Biochem Biophys Res Commun. 373:670–674. 2008. View Article : Google Scholar : PubMed/NCBI
39	Rayner KJ, Suárez Y, Dávalos A, Parathath S, Fitzgerald ML, Tamehiro N, Fisher EA, Moore KJ and Fernández-Hernando C: MiR-33 contributes to the regulation of cholesterol homeostasis. Science. 328:1570–1573. 2010. View Article : Google Scholar : PubMed/NCBI
40	Maxwell KN and Breslow JL: Adenoviral-mediated expression of Pcsk9 in mice results in a low-density lipoprotein receptor knockout phenotype. Proc Natl Acad Sci USA. 101:7100–7105. 2004. View Article : Google Scholar : PubMed/NCBI
41	Abifadel M, Varret M, Rabès JP, Allard D, Ouguerram K, Devillers M, Cruaud C, Benjannet S, Wickham L, Erlich D, et al: Mutations in Pcsk9 cause autosomal dominant hypercholesterolemia. Nat Genet. 34:154–156. 2003. View Article : Google Scholar : PubMed/NCBI
42	Krol J, Loedige I and Filipowicz W: The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 11:597–610. 2010.PubMed/NCBI
43	Kato M, de Lencastre A, Pincus Z and Slack FJ: Dynamic expression of small non-coding RNAs, including novel microRNAs and piRNAs/21U-RNAs, during Caenorhabditis elegans development. Genome Biol. 10:R542009. View Article : Google Scholar : PubMed/NCBI
44	Roglans N, Peris C, Verd JC, Alegret M, Vázquez M, Sánchez RM and Laguna JC: Increase in hepatic expression of SREBP-2 by gemfibrozil administration to rats. Biochem Pharmacol. 62:803–809. 2011. View Article : Google Scholar
45	Horton JD and Shimomura I: Sterol regulatory element-binding proteins: Activators of cholesterol and fatty acid biosynthesis. Curr Opin Lipidol. 10:143–150. 1999. View Article : Google Scholar : PubMed/NCBI
46	Horton JD: Sterol regulatory element-binding proteins: Transcriptional activators of lipid synthesis. Biochem Soc Trans. 30:1091–1095. 2002. View Article : Google Scholar : PubMed/NCBI