电针“足三里、三阴交”对脾气虚大鼠铁代谢影响的研究
发布时间：2018-08-13 21:22
【摘要】：目的:此次以脾气虚模型大鼠为实验动物,通过电针针刺“三阴交”和“足三里”穴对其进行治疗。以血清铁(SI)、铁蛋白(Fe)、总铁结合力(TIBC)、转铁蛋白(Tf)和转铁蛋白受体(Tf R)为观测目标,比较实验前后的数值变化,借此研究针刺治疗脾气虚方法和作用机制,同时尝试在分子生物学角度解释脾气虚时机体的病理变化。材料与方法:准备SD大鼠40只,然后随机分配到“正常组”、“模型组”、“经穴组”、“非经非穴组”四组。经由辽宁中医药大学实验动物中心对以上4组大鼠进行适应性饲养3天之后,对“模型组、经穴组、非经非穴组”进行为期2周的脾气虚造模,其造模因素为“过度疲劳,饮食不节”。完成造模后,对不同组进行不同方法的实验。每日对“正常组”进行正常喂养,不造模且不捆绑,不进行电针针刺治疗。每日上午继续对“模型组”进行造模,下午进行捆绑,时间为20min,不进行电针针刺治疗。每日上午继续对“经穴组”进行造模,下午捆绑后再用接电针仪的毫针分别刺入大鼠双侧的“三阴交”和“足三里”穴。电针参数选择疏密波,针刺强度以大鼠肌肉或针柄微颤为准,持续刺激20分钟,针刺深度3-5毫米。每日上午继续对“非经非穴组”进行造模,下午捆绑后用接电针仪的毫针刺入自定的非经非穴处,电针参数选择疏密波与“经穴组”一致,针刺强度以大鼠肌肉或针柄微颤为准,持续刺激20分钟,针刺深度3-5毫米。实验结束后,分别采集大鼠血清和肝脏细胞。用酶联免疫吸附法(ELISA方法)测定各组大鼠血清中血清铁含量、铁蛋白含量,计算总铁结合力。肝细胞中转铁蛋白m RNA和转铁蛋白受体m RNA的表达则通过采用荧光定量PCR方法进行检测。并采用统计软件spss19.0对所有实验所得数据予以分析。结果:1.电针针刺双侧“足三里”、“三阴交”穴后,脾气虚大鼠血清铁(SI)含量的变化和意义在大鼠血清铁含量上,正常组、经穴组间不具有统计学差异(P0.05),但与模型组相比,二者均呈明显下降趋势,差异具有统计学意义,(P值均0.01),模型组与非经非穴组比较无统计学意义(P0.05)。2.电针针刺双侧“足三里”、“三阴交”穴后,脾气虚大鼠铁蛋白(Fe)含量的变化和意义正常组大鼠铁蛋白含量与模型组相比显著升高(P0.01),经穴组铁蛋白含量与模型组相比显著升高(P0.01),经穴组与正常组相比无统计学意义(P0.05),模型组与非经非穴组比较无统计学意义(P0.05)3.电针针刺双侧“足三里”、“三阴交”穴后,脾气虚大鼠总铁结合力(TIBC)的变化和意义模型组大鼠血清总铁结合力与正常组相比显著降低(P0.01),经穴组总铁蛋白结合力与模型组相比显著升高(P0.01),经穴组与正常组相比无统计学意义(P0.05),模型组与非经非穴组比较无统计学意义(P0.05)。4.电针针刺双侧“足三里”、“三阴交”穴后,脾气虚大鼠转铁蛋白m RNA的表达量影响模型组大鼠转铁蛋白m RNA的表达量与正常组相比显著降低(P0.01),经穴组血清转铁蛋白m RNA的表达量与模型组相比显著升高(P0.01),经穴组与正常组相比无统计学意义(P0.05),模型组与非经穴组比较无统计学意义(P0.05)。5.电针针刺双侧“足三里”、“三阴交”穴后,脾气虚大鼠转铁蛋白受体m RNA的表达量影响模型组大鼠转铁蛋白受体m RNA的表达量与正常组相比显著降低(P0.01),经穴组血清转铁蛋白受体m RNA的表达量与模型组相比显著升高(P0.01),经穴组与正常组相比无统计学意义(P0.05),模型组与非经穴组比较无统计学意义(P0.05)。结论:1.当大鼠体内发生脾气虚时,机体内的铁代谢出现异常。2.电针针刺“足三里”和“三阴交”穴可以起到益气健脾的功效,在治疗脾气虚的同时也可以改善和调节脾气虚大鼠体内的铁代谢。3.脾主升清、脾主统血功能很可能是通过转铁蛋白-转铁蛋白受体系统(TTRS)来表现的。4.脾气虚的病理本质与铁代谢异常有着极大的关联。
[Abstract]:Objective: To study the therapeutic effect of acupuncture on spleen of rats with deficiency of spleen-qi (SQD) treated by Electroacupuncture at "Sanyinjiao" and "Zusanli" points. Serum iron (SI), ferritin (Fe), total iron binding capacity (TIBC), transferrin (Tf) and transferrin receptor (Tf R) were observed and compared before and after the experiment. Materials and Methods: Forty SD rats were prepared and randomly assigned to four groups: normal group, model group, acupoint group and non-acupoint group. After 3 days of adaptive feeding, the rats in the model group, the acupoint group and the non-acupoint group were modeled with Spleen-Qi Deficiency for 2 weeks. The modelling factors were "excessive fatigue and poor diet". Acupuncture therapy.The model group was made in the morning and bundled in the afternoon for 20 minutes without electroacupuncture.The model group was made in the morning and then bundled in the afternoon.The filiform needles of the electroacupuncture apparatus were used to puncture the "Sanyinjiao" and "Zusanli" points on both sides of the rats.The parameters of electroacupuncture were selected. Needling intensity should be based on muscle or needle handle tremor of rats. The needling depth should be 3-5 mm. The model of "non-meridian non-acupoint group" should be made every morning. After binding in the afternoon, the needle of electroacupuncture apparatus should be inserted into the non-meridian non-acupoint group. The parameters of electroacupuncture should be the same as that of "meridian point group". Rat serum and liver cells were collected after the experiment. Serum iron content, ferritin content and total iron binding capacity were determined by enzyme linked immunosorbent assay (ELISA), and transferrin m RNA and transferrin receptor in liver cells were measured. The expression of body m RNA was detected by fluorescence quantitative PCR, and all the data were analyzed by statistical software SPSS 19.0. Results: 1. After electroacupuncture at Zusanli and Sanyinjiao points, the changes and significance of serum iron (SI) content in rats with spleen-qi deficiency were found in normal group, meridian group. There was no statistical difference between the two groups (P 0.05), but compared with the model group, both showed a significant downward trend, the difference was statistically significant (P 0.01). There was no significant difference between the model group and the non-meridian acupoint group (P 0.05). Ferritin content in normal group was significantly higher than that in model group (P 0.01). Ferritin content in acupoint group was significantly higher than that in model group (P 0.01). There was no significant difference between acupoint group and normal group (P 0.05). There was no significant difference between model group and non-acupoint group (P 0.05). 3. Changes of total iron binding capacity (TIBC) in spleen-qi deficiency rats and its significance The total iron binding capacity in serum of model group was significantly lower than that of normal group (P 0.01). The total ferritin binding capacity in acupoint group was significantly higher than that of model group (P 0.01). There was no significant difference between acupoint group and normal group (P 0.05). There was no statistical significance between model group and non-acupoint group. After electroacupuncture at Zusanli and Sanyinjiao points, the expression of transferrin m RNA in model group was significantly lower than that in normal group (P 0.01). The expression of transferrin m RNA in serum of acupoint group was significantly higher than that of model group (P 0.01). There was no significant difference between the normal group (P 0.05), the model group and the non-meridian group (P 0.05). 5. After electroacupuncture at Zusanli and Sanyinjiao, the expression of transferrin receptor m RNA in spleen-qi deficiency rats was significantly lower than that in the normal group (P 0.01). The expression of serum transferrin receptor m RNA in acupoint group was significantly higher than that in model group (P 0.01), and there was no significant difference between acupoint group and normal group (P 0.05). There was no significant difference between model group and non-acupoint group (P 0.05). Conclusion: 1. Iron metabolism was abnormal in rats with Spleen-Qi deficiency. "Sanyinjiao" and "Sanyinjiao" acupoints can play the role of Invigorating Qi and strengthening spleen, in the treatment of spleen-qi deficiency, but also can improve and regulate iron metabolism in rats with Spleen-Qi deficiency. A great correlation.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245
[Abstract]:Objective: To study the therapeutic effect of acupuncture on spleen of rats with deficiency of spleen-qi (SQD) treated by Electroacupuncture at "Sanyinjiao" and "Zusanli" points. Serum iron (SI), ferritin (Fe), total iron binding capacity (TIBC), transferrin (Tf) and transferrin receptor (Tf R) were observed and compared before and after the experiment. Materials and Methods: Forty SD rats were prepared and randomly assigned to four groups: normal group, model group, acupoint group and non-acupoint group. After 3 days of adaptive feeding, the rats in the model group, the acupoint group and the non-acupoint group were modeled with Spleen-Qi Deficiency for 2 weeks. The modelling factors were "excessive fatigue and poor diet". Acupuncture therapy.The model group was made in the morning and bundled in the afternoon for 20 minutes without electroacupuncture.The model group was made in the morning and then bundled in the afternoon.The filiform needles of the electroacupuncture apparatus were used to puncture the "Sanyinjiao" and "Zusanli" points on both sides of the rats.The parameters of electroacupuncture were selected. Needling intensity should be based on muscle or needle handle tremor of rats. The needling depth should be 3-5 mm. The model of "non-meridian non-acupoint group" should be made every morning. After binding in the afternoon, the needle of electroacupuncture apparatus should be inserted into the non-meridian non-acupoint group. The parameters of electroacupuncture should be the same as that of "meridian point group". Rat serum and liver cells were collected after the experiment. Serum iron content, ferritin content and total iron binding capacity were determined by enzyme linked immunosorbent assay (ELISA), and transferrin m RNA and transferrin receptor in liver cells were measured. The expression of body m RNA was detected by fluorescence quantitative PCR, and all the data were analyzed by statistical software SPSS 19.0. Results: 1. After electroacupuncture at Zusanli and Sanyinjiao points, the changes and significance of serum iron (SI) content in rats with spleen-qi deficiency were found in normal group, meridian group. There was no statistical difference between the two groups (P 0.05), but compared with the model group, both showed a significant downward trend, the difference was statistically significant (P 0.01). There was no significant difference between the model group and the non-meridian acupoint group (P 0.05). Ferritin content in normal group was significantly higher than that in model group (P 0.01). Ferritin content in acupoint group was significantly higher than that in model group (P 0.01). There was no significant difference between acupoint group and normal group (P 0.05). There was no significant difference between model group and non-acupoint group (P 0.05). 3. Changes of total iron binding capacity (TIBC) in spleen-qi deficiency rats and its significance The total iron binding capacity in serum of model group was significantly lower than that of normal group (P 0.01). The total ferritin binding capacity in acupoint group was significantly higher than that of model group (P 0.01). There was no significant difference between acupoint group and normal group (P 0.05). There was no statistical significance between model group and non-acupoint group. After electroacupuncture at Zusanli and Sanyinjiao points, the expression of transferrin m RNA in model group was significantly lower than that in normal group (P 0.01). The expression of transferrin m RNA in serum of acupoint group was significantly higher than that of model group (P 0.01). There was no significant difference between the normal group (P 0.05), the model group and the non-meridian group (P 0.05). 5. After electroacupuncture at Zusanli and Sanyinjiao, the expression of transferrin receptor m RNA in spleen-qi deficiency rats was significantly lower than that in the normal group (P 0.01). The expression of serum transferrin receptor m RNA in acupoint group was significantly higher than that in model group (P 0.01), and there was no significant difference between acupoint group and normal group (P 0.05). There was no significant difference between model group and non-acupoint group (P 0.05). Conclusion: 1. Iron metabolism was abnormal in rats with Spleen-Qi deficiency. "Sanyinjiao" and "Sanyinjiao" acupoints can play the role of Invigorating Qi and strengthening spleen, in the treatment of spleen-qi deficiency, but also can improve and regulate iron metabolism in rats with Spleen-Qi deficiency. A great correlation.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245
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