| 白 妍,董兆玲,卢金荣,林玉杰,肖 艳,李 元,秦正巍,王 顺."调神畅志"针刺法对帕金森伴便秘模型大鼠结肠组织5-HT4R及CaM-MLCK信号通路影响的研究[J].,2024,(3):410-414 |
| "调神畅志"针刺法对帕金森伴便秘模型大鼠结肠组织5-HT4R及CaM-MLCK信号通路影响的研究 |
| Study on the Influence of Acupuncture Method of "Tiaoshenchangzhi"Acupuncture on the 5-HT4R and CaM-MLCK Signaling Pathway in the Colon Tissue of Rats with Parkinson's Constipation Model |
| 投稿时间：2023-03-27 修订日期：2023-05-01 |
| DOI：10.13241/j.cnki.pmb.2024.03.003 |
| 中文关键词: 针刺 调神畅志法 帕金森伴便秘 5-羟色胺4受体 钙调蛋白-肌球蛋白轻链激酶信号通路信号通路 |
| 英文关键词: Acupuncture Tiaoshenchangzhi method Parkinson's disease with constipation 5-HT4R CaM-MLCK signaling pathway |
| 基金项目:黑龙江省中医药科研项目（ZHY19-070） |
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| 中文摘要: |
| 摘要 目的：探讨"调神畅志"针刺法对帕金森伴便秘(Parkinson's disease with constipation)模型大鼠结肠组织中5-羟色胺4受体（5-HT4R）及钙调蛋白（CaM）-肌球蛋白轻链激酶（MLCK）信号通路影响的研究，并探讨其可能作用机制。方法：采用随机数字表将60只大鼠分为空白组、假手术组、模型组、西药组、常规针刺组、调神畅志针刺组，每组10只。模型组、常规针刺组、调神畅志针刺组颈背部皮下注射鱼藤酮的方法制造PDD大鼠模型,并用阿扑吗啡（APO）诱导检测。常规针刺组选取天枢、足三里、上巨虚及舞蹈震颤控制区，接电针，连续波频率15Hz，刺激强度2 mA，留针30分钟，1日1次，连续4周；调神畅志针刺组：选取王顺教授 " 调神畅志 " 三六九针法，百会透太阳、中脘、气海、足三里、太冲，采用连续波，头针频率45Hz，腹针频率30Hz，肢体针频率15Hz（体现调神畅志三六九针法的头针重、腹针中、四肢针轻的量化刺激），刺激强度约2 mA，留针30 min，1日1次，连续4周；西药组采用美多芭、莫沙必利每日灌胃，连续治疗4周；模型组、假手术组：每日2 mL生理盐水灌胃，连续4周；空白组不予处理。采用Western blot法检测各组近端结肠组织中5-HT4R、CaM及MLCK蛋白表达以及含量水平。结果：与空白组相比，模型组大鼠近端结肠组织中5-HT4R、CaM及MLCK蛋白表达水平显著降低(P<0.01)，与模型组比较，三个治疗组（西药组、常规针刺组、调神畅志针刺组）近端结肠组织5-HT4R、CaM及MLCK蛋白表达水平显著升高(P<0.01)。与常规针刺组相比，调神畅志组5-HT4R、CaM及MLCK蛋白表达水平明显升高(P<0.05)。结论：针刺可改善PD伴便秘大鼠模型便秘症状，且调神畅志针刺组优于常规针刺组，其作用机理可能通过调节大鼠结肠组织中5-HT4R蛋白表达以及激活CaM-MLCK信号通路来实现的。 |
| 英文摘要: |
| ABSTRACT Objective: To discover the effects of "Tiaoshenchangzhi" acupuncture on 5-HT4R and CaM-myosin in the colon tissue of rats with Parkinson's disease with constipation, and to examine their possible mechanisms of action. light link protein kinase (MLCK) signaling pathway in the colon tissue of rats with Parkinson's disease with restriction, and to explore their possible mechanisms of action. Methods: 60 rats were classified into the blank group, the sham operation group, the modelling group, the western medicine group, the regular acupuncture group and the "Tiaoshenchangzhi" acupuncture group using a casual number table, with 10 rats in each group. The rat model of Parkinson's disease(PD) with constipation was made by subcutaneous injection of rotenone into the neck and back of model group, routine acupuncture group and Tiaoshenchangzhi acupuncture group, and detected by APO induction. In the conventional acupuncture group, Tianshu, Zusanli, Shangjuxu and dance tremor control areas were selected and electroacupuncture was performed with continuous wave frequency of 15Hz and stimulation intensity of 2 mA. The needles were kept for 30 minutes, once a day, for consecutive 4 weeks. Tiaoshenchangzhi acupuncture group: Professor Wang Shun's "TiaoshenChangzhi" thirty-nine needle method was selected, Baihui penetrating the Taiyang, Zhongwuan, Qihai, Zusanli and Taichong. Soil continuous wave was used, with the frequency of head needle 45Hz, abdominal needle 30Hz and limb needle 15Hz(To reflect the quantitative stimulation of the weight of the head needle, the weight of the abdomen needle and the light of the limbs needle of The 369 acupuncture method of Tiaoshenchangzhi) > the stimulation intensity was about 2mA, and the needle was left for 30 min, once a day, for consecutive 4 weeks; The western medicine group received metopar and mosapride daily gavage for 4 weeks. Model group and sham operation group: gavage of 2 mL normal saline daily for 4 consecutive weeks; Blank groups are not processed. Western blot was used to detect the expression and content of 5-HT4R, CaM and MLCK protein in the proximal colon tissues of each group. Results: Compared with the blank group, the expression of 5-HT4R, CaM and MLCK protein in the proximal colon tissue of model group were significantly decreased(P<0.01). Compared with the model group, the expression of 5-HT4R, CaM and MLCK protein in the proximal colon tissue of three treatment groups (Western medicine group, routine acupuncture group and Tiaoshenchangzhi acupuncture group)were significantly decreased Elevated(P<0.01). Compared with conventional acupuncture group, the expression of 5-HT4R, CaM and MLCK protein in Tiaoshenchangzhi group was significantly increased(P<0.05). Conclusion: Acupuncture can ameliorate the symptoms of constipation in the rat model of PD with constraints, and the "Tiaoshenchangzhi"acupuncture group was better than the conventional acupuncture group. The mechanism of action may be realized by adjusting the 5-HT4R protein expression and activating the CaM-MLCK signaling pathway in the rat colon tissue. |
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