低频耳电针调节糖尿病胃轻瘫大鼠胃动力及共病抑郁机制的实验研究
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南京医科大学第一附属医院针灸科

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江苏省卫生健康委科研立项项目重点项目(NO.ZD2021032);江苏省第三批省名老中医药专家传承工作室建设项目(苏中医科教﹝2019﹞10号)


Experimental Study on the Mechanism of Low-frequency Auricular Electroacupuncture Regulating Gastric Motility and Comorbid Depression in Diabetic Gastroparesis Rats
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    摘要:

    目的:观察耳电针对糖尿病胃轻瘫大鼠行为学、胃排空率和胃窦组织中酪氨酸激酶受体(c-Kit)和促肾上腺皮质释放激素因子(Corticotropin Releasing Factor, CRF)表达的影响,探讨耳电针改善糖尿病胃轻瘫大鼠胃排空和抑郁样行为的作用机制。方法:将36只SPF级SD大鼠随机分为空白对照组、模型组、耳电针组和假耳针组,每组9只。空白对照组腹腔注射柠檬酸钠缓冲液,其余三组腹腔注射链脲佐菌素(Streptozotocin, STZ)-柠檬酸钠缓冲液,随后继续饲养6周后开始治疗,耳电针组电针双侧耳穴“胃”,每日1次,每周5日,连续2周;假耳针组针刺耳垂,不加电。治疗结束后,进行行为学以及胃排空率检测,qRT-PCR检测胃窦组织CRF、c-Kit mRNA 表达,ELISA 检测胃窦组织CRF、c-Kit 蛋白表达 。结果:旷场实验中模型组大鼠进入中央区域次数(CE)、进入中央区域时间百分比(CT%)和进入中央区域距离百分比(CD%)较空白对照组均明显降低(均P<0.05),耳电针组CE、CT%和CD%值较模型组明显升高(均P<0.05);强迫游泳实验中模型组大鼠漂浮时间百分比(FT%)较空白对照组明显升高(P<0.05),耳电针组FT%值较模型组明显降低(P<0.05);与空白对照组相比,模型组大鼠胃排空率显著降低,胃窦中CRF的mRNA和蛋白表达水平明显升高(P<0.05),c-Kit的mRNA和蛋白表达水平明显降低(P<0.05);与模型组和假耳针组比较,耳针组大鼠胃排空率显著升高,胃窦中CRF的mRNA和蛋白表达水平明显降低(P<0.05),c-Kit的mRNA和蛋白表达水平明显升高(P<0.05)。结论:低频耳电针可以改善糖尿病胃轻瘫大鼠的胃动力与抑郁样行为,其机制可能与调节胃窦组织中相关因子的表达有关。

    Abstract:

    Objective To observe the effects of auricular electroacupuncture on behavior, gastric emptying rate and expression of tyrosine kinase receptor (c-Kit) and corticotropin releasing factor (CRF) in gastric sinus tissues of rats with diabetic gastroparesis (DGP), and to investigate the mechanism of auricular electroacupuncture to improve gastric emptying and depression-like behavior in DGP rats. Methods: Thirty-six SPF-grade SD rats were randomly divided into control group, model group, auricular electroacupuncture group and sham auricular acupuncture group, with 9 rats in each group. The control group was injected intraperitoneally with sodium citrate buffer, and the remaining three groups were injected with streptozotocin (STZ)-sodium citrate buffer. The treatment was continued for 6 weeks, followed by electro-acupuncture of bilateral auricular points "stomach" once a day for 5 days a week for 2 weeks in the auricular electroacupuncture group, and acupuncture of the earlobe without electricity in the sham auricular acupuncture group. After the treatment, the behavioral and gastric emptying rates were measured and the protein and mRNA levels of CRF and c-kit were respectively detected by WB and qRT-PCR in gastric sinus tissue. Results: In the Open Field test, the entries into the central area (CE), the percentage of time entering the central area (CT%) and the percentage of distance entering the central area (CD%) were significantly lower in the model group compared with the control group (all P<0.05), and CE, CT% and CD% were significantly higher in the auricular electroacupuncture group compared with the model group (all P<0.05); in the Forced Swimming test, the percentage of floating time (FT%) was significantly higher in the model group compared with the control group (P<0.05), and FT% were significantly lower in the auricular electroacupuncture group compared with the model group (P<0.05); compared with the control group, the gastric emptying rate of rats in the model group was significantly lower, and the mRNA and protein expression levels of CRF in the gastric sinus were significantly higher (P<0.05), and the mRNA and protein expression levels of c-Kit were significantly lower (P<0.05); compared with the model group and the sham auricular acupuncture group, the gastric emptying rate of rats in the auricular acupuncture group was significantly higher, and the mRNA and protein expression levels of CRF in the gastric sinus were significantly lower (P<0.05), and the mRNA and protein expression levels of c-Kit were significantly higher (P<0.05). Conclusion: Low-frequency auricular electroacupuncture can improve gastric motility and depression-like behavior in diabetic gastroparesis rats, and its mechanism may be related to regulating the expression of related factors in gastric antrum tissue.

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  • 收稿日期:2023-02-10
  • 最后修改日期:2023-04-10
  • 录用日期:2023-05-23
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