A protocol of histone modification-based mechanistic study of acupuncture in patients with stable angina pectoris.
BACKGROUND
Angina pectoris (Angina) is a medical condition related to myocardial ischemia. Although acupuncture has been widely accepted as a clinical approach for angina, there is no sufficient evidence of its effectiveness against this syndrome, and its mechanisms have not yet been well elucidated. We develop this protocol to confirm the clinical efficacy of electro-acupuncture on stable angina pectoris by needling on acupoint Neiguan (PC6). Furthermore, we employ high-throughput sequencing technology to investigate the gene expression profiling and determine involvement of histone modifications in the regulation of genes after electro-acupuncture treatment.


METHODS
A randomized, controlled, double-blinded (assessor and patients) trial will be carried out. Sixty participants will be randomly assigned to two acupuncture treatment groups and one control group in a 1:1:1 ratio. Participants in acupuncture groups will receive 12 sessions of electro-acupuncture treatment across 4 weeks, followed by a 12-week randomization period. The acupuncture groups are divided into Neiguan (PC6) on Pericardium Meridian of Hand-jueyin or a non-acupoint. The primary clinical measure of effect is the frequency of angina attacks between these groups for four weeks after randomization. RNAs are extracted from peripheral neutrophils collected from all participants on day 0, day 30, and week 16, and are processed to RNA-Seq. We then investigate profiles of histone modifications by ChIP-Seq, for H3 Lysine 4 (H3K4me) and acetylation of H3 Lysine 27 (H3K27ac), in the presence or absence of acupuncture treatment.


CONCLUSIONS
This study determines the efficacy and mechanisms of electro-acupuncture on stable angina pectoris. We focus on effectiveness of acupuncture on alleviating symptoms of myocardial ischemia and the gene regulation and the chromatin remodeling marks, including H3K4me1, H3K4me2, and H3K27ac, which could be key factors for regulating gene expressions caused by electro-acupuncture treatment at Neiguan. This is the first genome-wide study of electro-acupuncture treatment in angina patients, and will provide valuable information for future studies in the fields of acupuncture and its underlying mechanisms. Fourteen patients have been recruited since recruitment opened in November of 2012. This study is scheduled to end in November of 2014.


BACKGROUND
ChiCTR-TRC-12002668.
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. 39725409@qq.com.
Ning
N
Wang
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. crazywind@vip.qq.com.
Sheng-Feng
SF
Lu
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. 1078137043@qq.com.
Hui
H
Chen
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. 353980182@qq.com.
Jian-Fei
JF
Wang
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. 53894289@qq.com.
Shu-Ping
SP
Fu
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. hucjhyl@126.com.
Chen-Jun
CJ
Hu
School of Acupuncture and Tuina, Chengdu University of Traditional Chinese Medicine, 610075, Chengdu, Sichuan, China. 657172145@qq.com.
Yi
Y
Yang
School of Acupuncture and Tuina, Chengdu University of Traditional Chinese Medicine, 610075, Chengdu, Sichuan, China. acuresearch@126.com.
Fan-Rong
FR
Liang
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, 210023, Nanjing, China. zhubm64@hotmail.com.
Bing-Mei
BM
Zhu
D015669:Acupuncture Points; D000328:Adult; D000368:Aged; D000787:Angina Pectoris; D060050:Angina, Stable / Q000235:genetics / Q000628:therapy*; D042002:Chromatin Assembly and Disassembly; D002985:Clinical Protocols; D015671:Electroacupuncture; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D055106:Genome-Wide Association Study; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D012107:Research Design
D006657:Histones; D008239:Lysine