穴位埋線與電針太衝穴治療保護自發性高血壓大鼠心臟效果及其分子機轉探討

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题名:	穴位埋線與電針太衝穴治療保護自發性高血壓大鼠心臟效果及其分子機轉探討 Investigation the Cardio-protective Effects and molecular mechanisms of Catgut Embedding and Electro Acupuncture Therapy in TaiChong point of Spontaneous Hypertensive Rats
作者:	陳怡吏
贡献者:	針灸研究所碩士班
关键词:	電針;太沖穴;高血壓;自發性高血壓大鼠;心肌肥大;凋亡;埋線 electro-acupuncture;catgut embedding;Taichongpoint(LR-3);SHR;cardiac hypertrophy;apoptosis;catgut embedding;Taichongpoint(LR-3)
日期:	2011-07-08
上传时间:	2011-10-17 16:52:12 (UTC+8)
出版者:	中國醫藥大學
摘要:	(PART 1) 背景：高血壓是因為全身動脈血壓升高的一種疾病。約90-95 ％的高壓病人找不到病因，是自發性的，所以被稱為“原發性高血壓“，它可以引起嚴重的心、腦、腎功能障礙。持久性高血壓有可能造成心肌纖維化、左心室肥大及心臟的細胞凋亡而使壽命縮短。在臨床上常有服西藥控制中的病人抱怨服藥所帶來的副作用。一個大量的臨床研究顯示，針灸太沖穴（LR-3）可以有效地降低血壓，可用於治療高血壓。目的：我們通過電針刺激肝經的太沖穴來觀察降血壓的效果及對訊息傳導路徑的影響，而針灸治療高血壓具有療效顯著、副作用小、操作簡便等特點，並可以改善高血壓引起的症狀，是臨床上治療高血壓的常用方法之一。根據研究獨取太沖穴治療肝陽上亢型高血壓其降壓效果良好，降壓幅度與針刺前血壓呈正相關，不良反應少。我們用自發性高血壓大鼠（SHR）的動物模型來研究電針的效果是否對自發性高血壓大鼠的心臟功能有正面的影響。方法：以8週大的雄性自發性高血壓大鼠隨機分為高血壓控制組 (SHR)、電針偽穴位組 (EAS)、電針治療組 (EAT)，進行3週的治療。並以8周大的雄性WKY (Wistar- Kyoto) 大鼠作為對照組。治療組大鼠在雙側太沖穴電針，並記錄各組大鼠的血壓變化。犧牲後取其心臟做切片，並觀察H & E、Masson’s trichrome 染色與TUNEL assay。游離出左心室並萃取蛋白質，進行西方墨點法，觀察SOD 1、eNOS、肥大與凋亡訊息路徑蛋白質的變化。結果與討論：治療第三天，EAT組大鼠的血壓即有明顯下降，包括收縮壓、平均血壓與舒張壓都顯著低於SHR與EAS組。到治療第19天時EAT組大鼠只有收縮壓仍顯著低於SHR與EAS組。顯示電針太沖穴能達到迅速降血壓的效果，但長期的連續刺激，其降血壓效果會逐漸減弱。從切片可以觀察到電針太沖穴可降低高血壓對心臟造成的肥大、凋亡和纖維化的程度。西方墨點法分析發現經過治療的大鼠，eNOS與SOD 1明顯增加，且肥大與凋亡路徑的蛋白質均明顯下降。 (PART Ⅱ ) 背景：持久性高血壓有可能造成心肌纖維化、左心室肥大及心臟的細胞凋亡而使壽命縮短。目的：我們從先前的實驗得知電針太沖穴能有效降低自發性高血壓大鼠的血壓，但長期的連續刺激會讓此效果逐漸減弱。因此我們設計一個治療時間較短的實驗，並加入埋線治療的方式一起討論。方法：以8週大的雄性自發性高血壓大鼠隨機分為高血壓控制組 (SHR)、電針偽穴位組 (EAS)、電針治療組 (EAT)，埋線偽穴位組 (CES)、埋線治療組 (CET)，進行1週的治療。並以8周大的雄性WKY (Wistar-Kyoto) 大鼠作為對照組。治療組大鼠在雙邊太沖穴電針/埋線，並記錄各組大鼠的血壓變化。犧牲後取其心臟做切片，並觀察H & E、Masson’s trichrome 染色與TUNEL assay。游離出左心室並萃取蛋白質，進行西方墨點法，觀察凋亡與抗凋亡訊息路徑蛋白質 and cardial hypertrophy and inflammation結果與討論：治療第三天，EAT與CET組大鼠的血壓即有明顯下降，包括收縮壓、平均血壓與舒張壓都顯著低於SHR與EAS組，且EAT組的血壓略低於CET組但兩組間無顯著差異。到治療第7天時EAT與CET組大鼠血壓仍持續下降，且CET組血壓已低於EAT組但兩組間無顯著差異。從切片可以觀察到電針/埋線太沖穴可降低高血壓對心臟造成的肥大、凋亡和纖維化。西方墨點法分析可發現經過治療的大鼠， SOD 1與抗凋亡路徑的蛋白質明顯增加，且肥大與凋亡路徑的蛋白質均明顯下降，且埋線對SOD 1與PI3K/pPI3K的誘導效果明顯優於電針。 (PART 1) BACKGROUND: Hypertension is a kind of disease which the systemic arterial blood pressure is elevated. About 90–95% of cases are termed "spontaneous hypertension", which refers to high blood pressure for no medical cause can be found. It can cause serious heart, brain and kidney dysfunction. Persistent hypertension may cause cardiac fibrosis, left ventricular hypertrophy and cardiac apoptosis, leaving shortened life expectancy. Lots of clinical experiences have shown that puncture Taichong point(LR-3)can effectively lower blood pressure. OBJECTIVE: We punctured LR-3 point to observe the effect of lowering blood pressure and to explore the molecular mechanism of acupuncture treatment for hypertension. Acupuncture treatment for hypertension has less side effect. This is the common point be used in clinic. According to the research, even using the single point of Taicong(LR-3) also can lower hyper-yang activity type hypertension well. We use SHR to make sure whether the effect of electro -acupuncture on the cardiac function of SHR has a positive impact. METHODS: A total of 12 rats with 8-weeks male spontaneously hypertensive rats (SHR)were randomized divided into the acupuncture and sham groups, with 6 animals in each group. Additional 6 male Wistar-Kyoto rats were served as the control group. Rats in the acupuncture group received electro-acupuncture at bilateral Taichong (LR 3) points. Stained with Masson trichrome to observe whether there is myocardial fibrosis.and to observe myocardial cell apoptosis by TUNEL level. The expression of SOD1, FAS, FADD, Bax, cytochromC, Gαq, eNOS, and calcineurin and BNP were detected by Western blot. RESULTS AND CONCLUSION: After treatment for three days, EAT group of blood pressure in rats that have decreased significantly, including systolic(BPs), diastolic(BPd) and mean blood pressure (BPm)were significantly lower than SHR and EAS group. At 19th day , the systolic blood pressure in EAT group was still significantly lower than SHR and EAS group. Electroacupuncture LR-3 has shown that the blood pressure was went down rapidly, but the long-term continuous stimulus, its antihypertensive effects would fade. Electroacupuncture LR-3 can reduce the damage of cardiac hypertrophy and apoptosis and fibrosis caused by hypertension. According to Western blot analysis, we found the eNOS and SOD 1 were increased. In another pathway,, the hypertrophy and apoptosis protein were decreased significantly. (PART Ⅱ ) Background: the persistent high blood pressure might cause myocardial fibrosis and left ventricular hypertrophy and apoptosis. Finally shorten the life. Objective: we learned from the previous experimental Taichong electroacupuncture is effective for reducing blood pressure in spontaneously hypertensive rats, but long-term stimulation will make the effect fade in a row. So we designed a short experimental treatment and join discussions. Methods: the 8-week-old male group randomly divided into Spontaneous hypertensive rat group (SHR), pseudo-point electroacupuncture Group (EAS), electro-acupuncture in the treatment group (EAT), pseudo-acupoint embedding Group (CES), embedding therapy group (CET), for 1 week of treatment. With the 8-week-old male WKY (Wistar-Kyoto) as a control group of rats. Treatment group we chose bilateral Taichongpoint(LR-3) for electro-acupuncture and catgut embedding, and recorded their blood pressure with tail cuff. . After sacrifice we took its heart slices, and observed H&E, Masson ' s trichrome staining with TUNEL assay. Free out of the left ventricle and extraction of proteins, by Western blot to observe the proteins of anti-apoptosis and pro-apoptosis proteins and the proteins about hypertrophy or inflammation. Results and discussion: the treatment of the third day, the blood pressure of rats with has decreased significantly, including systolic, diastolic and mean blood pressure in EAT and CET group were significantly lower than SHR and EAS group, and the blood pressure in EAT group was slightly lower than the CET group , but no significant difference between the two groups. At the treatment of the 7th day EAT blood pressure in CET Group continued to decline, and in CET group blood pressure was less than EAT group but no significant difference between the two groups. We observed from the heart tissue biopsy proofed that the electro-acupuncture or catgut embedding on LR-3 can prevent cardiac hypertrophy and apoptosis and fibrosis from hypertension. Western blot analysis was found that the SOD 1 in CET group was significantly increased and apoptotic protein and hypertrophy and apoptosis protein were decreased significantly, and SOD 1 and the induction of effect of PI3K/pPI3K in CET group were better than EAT group.
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