Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 734-744.DOI: 10.19852/j.cnki.jtcm.20240610.004
• Original articles • Previous Articles Next Articles
CHEN Yonglin, OUYANG Ling, MENG Lingling, WU Bufan, PENG Rou, LIU Sitong, HOU Dan, WANG Yaling, JING Xinyue, LU Shengfeng(), FU Shuping()
Received:
2023-02-22
Accepted:
2023-07-20
Online:
2024-08-15
Published:
2024-06-10
Contact:
Prof. FU Shuping, Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing, 210023, China. Supported by:
CHEN Yonglin, OUYANG Ling, MENG Lingling, WU Bufan, PENG Rou, LIU Sitong, HOU Dan, WANG Yaling, JING Xinyue, LU Shengfeng, FU Shuping. Electroacupuncture ameliorates blood-brain barrier disruption after ischemic stroke through histone acetylation regulation at the matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 2 genes[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 734-744.
Figure 1 EA treatment reduced cerebral damage after I/R injury in rats A: the experimental timeline; B: Longa neurologic score 1 d after ischemia, n = 11; C: Longa neurologic score 3 d after ischemia, n = 7; D: representative photograph of Evans blue extravasation in the brains of rats 24 h after I/R injury. The blue area shows penetrating Evans blue, signifying BBB destruction; E: quantitative analysis of Evans blue penetration in each group 1 d after ischemic stroke, n = 7; F: Evans blue exudation 3 d after ischemia, n = 4. Sham group: sham operated; MCAO group: MCAO operated; MEA group: MCAO rats received 3 d of EA treatment; HATi group: MCAO rats received 3 d of EA treatment and a single 4 mg/kg intraperitoneal injection of HATi. I/R: ischemia/reperfusion; MCAO: middle cerebral artery occlusion; EA: electroacupuncture; HATi: histone acetyltransferase inhibitor; EB: Evans blue; RT-qPCR: real-time quantitative polymerase chain reaction; ChIP: chromatin immunoprecipitation; BBB: blood-brain barrier. Data were analyzed by one-way analysis of variance and are represented as the mean ± standard deviation. aP < 0.05 vs the sham group, bP < 0.05 the MCAO group vs the MEA group, cP < 0.05 the MEA group vs the HATi group.
Figure 2 EA treatment regulated the expression of MMP-9 and TIMP-1/2 after MCAO A: RT-qPCR shows the relative mRNA levels of MMP-9 in ischemic brain tissues 24 h after I/R injury, n = 4; B: RT-qPCR shows the relative mRNA levels of TIMP-1 in ischemic brain tissues 24 h after ischemia/reperfusion injury, n = 4; C: RT-qPCR shows the relative mRNA levels of TIMP-2 in ischemic brain tissues 24 h after I/R injury, n = 4. Sham group: sham operated; MCAO group: MCAO operated; MEA group: MCAO rats received 3 d of EA treatment; HATi group: MCAO rats received 3 d of EA treatment and a single 4 mg/kg intraperitoneal injection of HATi. MMP-9: matrix metalloproteinase 9; TIMP-1/2: tissue inhibitor of metalloproteinase 1/2; MCAO: middle cerebral artery occlusion; EA: electroacupuncture; HATi: histone acetyltransferase inhibitor; RT-qPCR: real-time quantitative polymerase chain reaction. Data were analyzed by one-way analysis of variance and are represented as the mean ± standard deviation. aP < 0.05, vs the sham group; bP < 0.05, the MCAO group vs the MEA group; cP < 0.05, the MEA group vs the HATi group.
Figure 3 EA treatment regulated H3K9ace occupancy at target gene promoters A: ChIP analysis of the enrichment of acetylated H3K9 on promoters of MMP-9 -21 bp - -177 bp region, n = 3 or 4; B: ChIP analysis of the enrichment of acetylated H3K9 on promoters of MMP-9 -678 bp - -886 bp region, n = 3 or 4; C: ChIP analysis of the enrichment of acetylated H3K9 on promoters of TIMP-1 -271 bp - -549 bp region, n = 3 or 4; D: ChIP analysis of the enrichment of acetylated H3K9 on promoters of TIMP-1 -580 bp - -822 bp region, n = 3 or 4. E: ChIP analysis of the enrichment of acetylated H3K9 on promoters of TIMP-2 -248 bp - -425 bp region, n = 3 or 4. F: ChIP analysis of the enrichment of acetylated H3K9 on promoters of TIMP-2 -34 bp - -150 bp region, n = 3 or 4. Sham group: sham operated; MCAO group: MCAO operated; MEA group: MCAO rats received 3 d of EA treatment; HATi group: MCAO rats received 3 d of EA treatment and a single 4 mg/kg intraperitoneal injection of HATi. MCAO: middle cerebral artery occlusion; EA: electroacupuncture; HATi: histone acetyltransferase inhibitor; H3K9ace: histone H3 lysine 9 acetylation; MMP-9: matrix metalloproteinase 9; TIMP-1: tissue inhibitor of metalloproteinase 1; TIMP-2: tissue inhibitor of metalloproteinase 2; ChIP: chromatin immunoprecipitation. Data were analyzed by one-way analysis of variance and are represented as the mean ± standard deviation. aP < 0.05 vs the sham group, bP < 0.05 the MCAO group vs the MEA group, cP < 0.05 the MEA group vs the HATi group.
Figure 4 EA treatment regulated H3K27ace occupancy at target gene promoters A: ChIP analysis of the enrichment of acetylated H3K27 on promoters of MMP-9 -21 bp - -177 bp region, n = 3 or 4; B: ChIP analysis of the enrichment of acetylated H3K27 on promoters of MMP-9 -678 bp - -886 bp region, n = 3 or 4; C: ChIP analysis of the enrichment of acetylated H3K27 on promoters of TIMP-1 -271 bp - -549 bp region, n = 3 or 4; D: ChIP analysis of the enrichment of acetylated H3K27 on promoters of TIMP-1 -580 bp - -822 bp region, n = 3 or 4; E: ChIP analysis of the enrichment of acetylated H3K27 on promoters of TIMP-2 -248 bp - -425 bp region, n = 3 or 4. F: ChIP analysis of the enrichment of acetylated H3K27 on promoters of TIMP-2 -34 bp - -150 bp region, n = 3 or 4. Sham group: sham operated; MCAO group: MCAO operated; MEA group: MCAO rats received 3 d of EA treatment; HATi group: MCAO rats received 3 d of EA treatment and a single 4 mg/kg intraperitoneal injection of HATi. MCAO: middle cerebral artery occlusion; EA: electroacupuncture; HATi: histone acetyltransferase inhibitor; H3K27ace: histone H3 lysine 27 acetylation; MMP-9: matrix metalloproteinase 9; TIMP-1: tissue inhibitor of metalloproteinase 1; TIMP-2: tissue inhibitor of metalloproteinase 2; ChIP: chromatin immunoprecipitation. Data were analyzed by one-way analysis of variance and are represented as the mean ± standard deviation. aP < 0.05 vs the sham group, bP < 0.05 the MCAO group vs the MEA group, cP < 0.05 the MEA group vs the HATi group.
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