電針“足三里、三陰交”對脾氣虛大鼠鐵代謝影響的研究
發(fā)布時間：2018-08-13 21:22
【摘要】：目的:此次以脾氣虛模型大鼠為實驗動物,通過電針針刺“三陰交”和“足三里”穴對其進行治療。以血清鐵(SI)、鐵蛋白(Fe)、總鐵結(jié)合力(TIBC)、轉(zhuǎn)鐵蛋白(Tf)和轉(zhuǎn)鐵蛋白受體(Tf R)為觀測目標,比較實驗前后的數(shù)值變化,借此研究針刺治療脾氣虛方法和作用機制,同時嘗試在分子生物學角度解釋脾氣虛時機體的病理變化。材料與方法:準備SD大鼠40只,然后隨機分配到“正常組”、“模型組”、“經(jīng)穴組”、“非經(jīng)非穴組”四組。經(jīng)由遼寧中醫(yī)藥大學實驗動物中心對以上4組大鼠進行適應(yīng)性飼養(yǎng)3天之后,對“模型組、經(jīng)穴組、非經(jīng)非穴組”進行為期2周的脾氣虛造模,其造模因素為“過度疲勞,飲食不節(jié)”。完成造模后,對不同組進行不同方法的實驗。每日對“正常組”進行正常喂養(yǎng),不造模且不捆綁,不進行電針針刺治療。每日上午繼續(xù)對“模型組”進行造模,下午進行捆綁,時間為20min,不進行電針針刺治療。每日上午繼續(xù)對“經(jīng)穴組”進行造模,下午捆綁后再用接電針儀的毫針分別刺入大鼠雙側(cè)的“三陰交”和“足三里”穴。電針參數(shù)選擇疏密波,針刺強度以大鼠肌肉或針柄微顫為準,持續(xù)刺激20分鐘,針刺深度3-5毫米。每日上午繼續(xù)對“非經(jīng)非穴組”進行造模,下午捆綁后用接電針儀的毫針刺入自定的非經(jīng)非穴處,電針參數(shù)選擇疏密波與“經(jīng)穴組”一致,針刺強度以大鼠肌肉或針柄微顫為準,持續(xù)刺激20分鐘,針刺深度3-5毫米。實驗結(jié)束后,分別采集大鼠血清和肝臟細胞。用酶聯(lián)免疫吸附法(ELISA方法)測定各組大鼠血清中血清鐵含量、鐵蛋白含量,計算總鐵結(jié)合力。肝細胞中轉(zhuǎn)鐵蛋白m RNA和轉(zhuǎn)鐵蛋白受體m RNA的表達則通過采用熒光定量PCR方法進行檢測。并采用統(tǒng)計軟件spss19.0對所有實驗所得數(shù)據(jù)予以分析。結(jié)果:1.電針針刺雙側(cè)“足三里”、“三陰交”穴后,脾氣虛大鼠血清鐵(SI)含量的變化和意義在大鼠血清鐵含量上,正常組、經(jīng)穴組間不具有統(tǒng)計學差異(P0.05),但與模型組相比,二者均呈明顯下降趨勢,差異具有統(tǒng)計學意義,(P值均0.01),模型組與非經(jīng)非穴組比較無統(tǒng)計學意義(P0.05)。2.電針針刺雙側(cè)“足三里”、“三陰交”穴后,脾氣虛大鼠鐵蛋白(Fe)含量的變化和意義正常組大鼠鐵蛋白含量與模型組相比顯著升高(P0.01),經(jīng)穴組鐵蛋白含量與模型組相比顯著升高(P0.01),經(jīng)穴組與正常組相比無統(tǒng)計學意義(P0.05),模型組與非經(jīng)非穴組比較無統(tǒng)計學意義(P0.05)3.電針針刺雙側(cè)“足三里”、“三陰交”穴后,脾氣虛大鼠總鐵結(jié)合力(TIBC)的變化和意義模型組大鼠血清總鐵結(jié)合力與正常組相比顯著降低(P0.01),經(jīng)穴組總鐵蛋白結(jié)合力與模型組相比顯著升高(P0.01),經(jīng)穴組與正常組相比無統(tǒng)計學意義(P0.05),模型組與非經(jīng)非穴組比較無統(tǒng)計學意義(P0.05)。4.電針針刺雙側(cè)“足三里”、“三陰交”穴后,脾氣虛大鼠轉(zhuǎn)鐵蛋白m RNA的表達量影響模型組大鼠轉(zhuǎn)鐵蛋白m RNA的表達量與正常組相比顯著降低(P0.01),經(jīng)穴組血清轉(zhuǎn)鐵蛋白m RNA的表達量與模型組相比顯著升高(P0.01),經(jīng)穴組與正常組相比無統(tǒng)計學意義(P0.05),模型組與非經(jīng)穴組比較無統(tǒng)計學意義(P0.05)。5.電針針刺雙側(cè)“足三里”、“三陰交”穴后,脾氣虛大鼠轉(zhuǎn)鐵蛋白受體m RNA的表達量影響模型組大鼠轉(zhuǎn)鐵蛋白受體m RNA的表達量與正常組相比顯著降低(P0.01),經(jīng)穴組血清轉(zhuǎn)鐵蛋白受體m RNA的表達量與模型組相比顯著升高(P0.01),經(jīng)穴組與正常組相比無統(tǒng)計學意義(P0.05),模型組與非經(jīng)穴組比較無統(tǒng)計學意義(P0.05)。結(jié)論:1.當大鼠體內(nèi)發(fā)生脾氣虛時,機體內(nèi)的鐵代謝出現(xiàn)異常。2.電針針刺“足三里”和“三陰交”穴可以起到益氣健脾的功效,在治療脾氣虛的同時也可以改善和調(diào)節(jié)脾氣虛大鼠體內(nèi)的鐵代謝。3.脾主升清、脾主統(tǒng)血功能很可能是通過轉(zhuǎn)鐵蛋白-轉(zhuǎn)鐵蛋白受體系統(tǒng)(TTRS)來表現(xiàn)的。4.脾氣虛的病理本質(zhì)與鐵代謝異常有著極大的關(guān)聯(lián)。
[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.
【學位授予單位】：遼寧中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：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.
【學位授予單位】：遼寧中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R245
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