Effects of miR-9 and Tetramethylpyrazine on Activation of Hepatic Stellate Cells

• Published in: Biological & pharmaceutical bulletin Vol. 38; no. 3; pp. 396 - 401
• Main Authors: Li, Shan-Gao, Zhou, Jun, Zhong, Ji-Hong, Cao, Hai-Jun, Zhu, Ling, Liu, Jun, Hu, Hua-Jun, Lv, Bin
• Format: Journal Article
• Language: English; Japanese
• Published: Japan The Pharmaceutical Society of Japan 01.03.2015; Pharmaceutical Society of Japan; Japan Science and Technology Agency
• Subjects: Actins - metabolism; Collagen Type I - metabolism; Gene Expression Regulation; hepatic stellate cell (HSC); Hepatic Stellate Cells - drug effects; Hepatic Stellate Cells - metabolism; hsa-miR-9; Humans; Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3); Janus Kinase 1 - metabolism; leptin; Leptin - metabolism; Ligusticum - chemistry; Liver Cirrhosis - metabolism; Liver Cirrhosis - prevention & control; MicroRNAs - genetics; MicroRNAs - metabolism; Phosphorylation; Phytotherapy; Plant Extracts - pharmacology; Plant Extracts - therapeutic use; Pyrazines - pharmacology; Pyrazines - therapeutic use; Real-Time Polymerase Chain Reaction; RNA, Messenger - metabolism; Signal Transduction; STAT3 Transcription Factor - metabolism; tetramethylpyrazine (TMP); Transfection; Up-Regulation
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.b14-00611

Micro-RNAs (miRNAs) are involved in regulation of the incidence and development of several hepatic diseases. Thus manipulating miRNAs may be a promising therapeutic strategy against these entities. In this study hepatic stellate cells (HSCs) were transfected with hsa-miR-9 or anti-hsa-miR-9, treated with tetramethylpyrazine (TMP), or subjected to treatment with TMP and hsa-miR-9 transfection (combined treatment group). Then, real-time polymerase chain reaction (PCR) was performed to measure mRNA levels of hsa-miR-9. Expression of hsa-miR-9 was highest in the combination treatment group compared with other groups, and significantly higher than TMP-treated and hsa-miR-9-transfected groups (both p<0.05). The anti-hsa-miR-9-transfected group expressed the lowest mRNA level of hsa-miR-9 with marked decrease versus control (p<0.05). Downstream factors that may be affected by miR-9 such as leptin, α-smooth muscle actin (SMA), and collagen I, as well as phosphorylation levels of Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) were investigated at the protein level. All these factors were regulated contrariwise to expression trends of hsa-miR-9, showing the lowest level in the combination treatment group and highest level in anti-hsa-miR-9-transfected group. These results suggest that both transfection of hsa-miR-9 and TMP can lead to upregulated endogenous expression of hsa-miR-9, inhibit activation of JAK1/STAT3 signal pathway induced by leptin, and lead to reduction of α-SMA and collagen I—thus impeding activation of HSC.