Zhao Jingming, Li Hui, Shi Chong, Yang Tiezheng, Xu Baoshi
Proctology Department, Affiliated Hospital of Changchun University of Chinese Medicine, Changchun 130021, China.
Geriatric Department, Affiliated Hospital of Changchun University of Chinese Medicine, Changchun 130021, China.
Evid Based Complement Alternat Med. 2020 Feb 25;2020:4956179. doi: 10.1155/2020/4956179. eCollection 2020.
Irritable bowel syndrome (IBS) is a chronic functional bowel disease characterized by abdominal pain and changes in bowel habits in the absence of organic disease. Electroacupuncture (EA) has been shown to alleviate visceral hypersensitivity (VH) in IBS rat models by inhibiting the activation of astrocytes in the spinal cord. However, the underlying molecular mechanisms mediated by P2Y receptor of this effect of electroacupuncture remain unclear.
To explore whether EA inhibits the activity of astrocytes in the spinal cord dorsal horn of rat with visceral hypersensitivity by inhibiting P2Y receptor and its downstream mitogen activated protein kinase/extracellular regulated kinase 1 (MAPK/ERK) pathway.
Ten-day-old Sprague-Dawley (SD) male rats were given an intracolonic injection of 0.2 ml of 0.5% acetic acid (AA) to establish a visceral hypersensitivity model. EA was performed at Zusanli (ST 36) and Shangjuxu (ST 37) at 100 Hz for 1.05 s and 2 Hz for 2.85 s alternately, pulse width for 0.1 ms, 1 mA, 30 min/d, once a day, for 1 week. Cytokines IL-6, IL-1, and TNF- were analyzed by ELISA. The expressions of the P2Y receptor and pERK1/2 were analyzed by Western Blot and real-time PCR in the model and EA treated animals to explore the molecular mechanism of EA in inhibiting the activity of spinal cord dorsal horn (L-S segment) astrocytes in rats with IBS visceral hypersensitivity.
EA significantly reduced the behavioral abdominal withdrawal reflex score (AWRs) of IBS rats with visceral hypersensitivity induced by AA. For comparison, intrathecal injection of astrocytes activity inhibitor fluorocitrate (FCA) also reduced visceral hypersensitivity in IBS rats. EA at Zusanli and Shangjuxu inhibited the mRNA and protein expression of the glial fibrillary acidic protein (GFAP) and in rat spinal cord and reduced the release of inflammatory cytokines IL-6, IL-1, and TNF- were analyzed by ELISA. The expressions of the P2Y receptor and pERK1/2 were analyzed by Western Blot and real-time PCR in the model and EA treated animals to explore the molecular mechanism of EA in inhibiting the activity of spinal cord dorsal horn (L-S segment) astrocytes in rats with IBS visceral hypersensitivity. , and TNF-g, 10 g, 10 .
EA inhibited astrocyte activity in the spinal cord dorsal horn of rat with IBS visceral hypersensitivity by inhibiting the P2Y receptor and its downstream, PKC, and MAPK/ERK1/2 pathways.
肠易激综合征（IBS）是一种慢性功能性肠病，其特征为腹痛和排便习惯改变，且无器质性疾病。电针已被证明可通过抑制脊髓星形胶质细胞的激活来减轻IBS大鼠模型中的内脏超敏反应（VH）。然而，电针这种作用由P2Y受体介导的潜在分子机制仍不清楚。
探讨电针是否通过抑制P2Y受体及其下游的丝裂原活化蛋白激酶/细胞外调节激酶1（MAPK/ERK）途径来抑制内脏超敏大鼠脊髓背角星形胶质细胞的活性。
给10日龄的Sprague-Dawley（SD）雄性大鼠结肠内注射0.2 ml 0.5%乙酸（AA）以建立内脏超敏模型。电针双侧足三里（ST 36）和上巨虚（ST 37），频率交替为100 Hz持续1.05 s和2 Hz持续2.85 s，脉冲宽度0.1 ms，强度1 mA，每天30 min，连续1周。通过酶联免疫吸附测定（ELISA）分析细胞因子白细胞介素-6（IL-6）、白细胞介素-1（IL-1）和肿瘤坏死因子-α（TNF-α）。采用蛋白质免疫印迹法（Western Blot）和实时荧光定量聚合酶链反应（real-time PCR）分析模型组和电针治疗组动物中P2Y受体和磷酸化细胞外调节蛋白激酶1/2（pERK1/2）的表达，以探讨电针抑制IBS内脏超敏大鼠脊髓背角（L4-S1节段）星形胶质细胞活性的分子机制。
电针显著降低了由AA诱导的内脏超敏IBS大鼠的行为学腹部回撤反射评分（AWRs）。作为对照，鞘内注射星形胶质细胞活性抑制剂氟代柠檬酸（FCA）也降低了IBS大鼠的内脏超敏反应。电针双侧足三里和上巨虚可抑制大鼠脊髓中胶质纤维酸性蛋白（GFAP）的mRNA和蛋白表达，并通过ELISA分析降低炎性细胞因子IL-6、IL-1和TNF-α的释放。采用Western Blot和real-time PCR分析模型组和电针治疗组动物中P2Y受体和pERK1/2的表达，以探讨电针抑制IBS内脏超敏大鼠脊髓背角（L4-S1节段）星形胶质细胞活性的分子机制。
电针通过抑制P2Y受体及其下游的蛋白激酶C（PKC）和MAPK/ERK1/2途径，抑制了IBS内脏超敏大鼠脊髓背角的星形胶质细胞活性。
