印尼草藥在自發性高血壓大鼠經由抑制氧化壓力的 抗高血壓和心血管保護作用

Anti-hypertensive and cardiovascular protective effects of Indonesian herbal medicine through oxidative stress inhibition in spontaneously hypertensive rats

Erna Sulistyowati(Erna Sulistyowati)

指導教授 ： 葉竹來

高雄醫學大學/醫學院/醫學研究所博士班/博士(2018年)

摘要

中文摘要
高血壓是世界上許多高致死率的心血管疾病之一，現今有許多研究證實高血壓造成心血管損傷的主因是和氧化壓力息息相關。在氧化自由基訊息途徑上，NADPH氧化酶 (NOXs) 對於高血壓疾病扮演一個重要的調節角色，並且透過活化NF-B造成心血管損傷。傳統印尼藥物中利用雷公根、駁骨草和白茅煎煮而成的複方 (CJID) 能有效治療高血壓。然而，CJID預防高血壓引起的左心室肥大和血管重塑其機轉尚未明確。因此本研究目的探討CJI 在自發性高血壓大鼠 (SHRs) 模式中其抗高血壓和保護心血管作用。
本實驗設計使用自發性高血壓大鼠(SHRs)和血壓正常大鼠(WKY)分配成兩組，再隨機分配為CJID治療組和控制組。持續五週給予CJID後進行實驗，每週使用尾套法測得心臟收縮壓和心跳數率，並使用心臟超音波評估左心室功能。取大鼠血清、左心室和胸主動脈評估氧化還原恆定狀態。測量過氧化物和過氧化氫的產生量變化，運用免疫組織化學染色和免疫螢光分析左心室和胸主動脈中的iNOS、NFB-p65、NFB inhibitor alpha/IBα、p-IBα、NOX1、NOX2及NOX4的蛋白表現量改變，在左心室的檢體組織測量SOD2和SOD3的蛋白表現量。
結果發現在SHR大鼠經由CJID治療後，心臟收縮壓和心跳數率明顯的減少；在心臟超音波的結果顯示，使用CJID有效地改善左心室的形態和功能。CJID可改善SHR大鼠血管重塑作用，如中膜厚度、彈性和膠原蛋白堆積，降低LDH和MDA濃度，以及增加SOD和NO的表現量；在SHR大鼠治療組的左心室和胸主動脈內過氧化物和過氧化氫濃度皆降低。在左心室和胸主動脈，CJID抑制NOX1、NOX2及NOX4的蛋白表現量，更進一步發現iNOS、p-IBα-IκBα比值和NFκB-p65的表現量也被減低，並增加心臟中SOD的表現量。
總結，CJID可以預防左心室肥大和血管重塑作用，藉由減少自由基產生並透過調控NOXs-ROS-NFB之訊息傳遞路徑。本研究揭示CJID有效預防高血壓所引起的心血管疾病，未來將需動物和臨床實驗去證實上述重要的發現結果。

關鍵字

印尼中草藥 ； 心血管損傷 ； 氧化壓力 ； NADPH氧化酶B細胞活化核轉錄因子κ ； 自發性高血壓大鼠

並列摘要

Hypertension is one of several risks leading to / causing mortality-related cardiovascular diseases in the world. A remarkable evidence denotes that hypertension is associated with oxidative stress which in turn causes cardiovascular injury. Major ROS-generating source NADPH oxidases (NOXs) play distinct roles in regulating ROS-mediated signaling in hypertension, and the up-regulation induce cardiovascular damage through NF-ĸB activation. Traditional Indonesian medicine Centella asiatica, Justicia gendarussa and Imperata cylindrica decoction (CJID) is known to be efficacious for hypertension. However, it remains unclear whether CJID prevents hypertension-induced left ventricular hypertrophy and vascular remodeling. This study purposed to investigate the antihypertension and cardiovascular protection effects of CJID in spontaneously hypertensive rats (SHRs).
Ten-week-old SHRs and normotensive Wistar Kyoto (WKY) rats were randomly allocated and divided into two groups: CJID-treated and control. After 5 weeks of treatment, all rats were sacrificed. Systolic blood pressure and heart rates were weekly measured using tail-cuff method. The left ventricle (LV) function and its performance were evaluated with echocardiography. The redox homeostasis was evaluated in the serum, LV and thoracic aorta (TA) as well. Superoxide and H2O2 generations were observed. Changes in the level of iNOS, NF-ĸB-p65, NF-ĸB inhibitor alpha/IĸBα, p-IĸBα and NOX1, NOX2, NOX4 in LV and TA were determined by immunohistochemistry and immunofluorescence analysis, respectively. The expressions of SOD2 and SOD3 were measured in the LV tissue.
We found that SBP and HR were significantly decreased in SHRs-treated group. Echocardiography showed that CJID significantly improved LV morphometry and function. Vascular remodeling such as media thickness, collagen and elastic accumulation were improved in CJID-treated group of SHRs. CJID attenuated lactate dehydrogenase (LDH) and malondialdehyde (MDA) levels increased SOD and NO levels. Superoxide and H2O2 generation was decreased both in LV and TA of SHRs-treated group. CJID caused the expressions of NOX1, NOX2, NOX4 in LV and TA to be suppressed. Furthermore, iNOS, ratio of p-IBα-IBα and NF-B-p65 expressions were decreased both in LV and TA. CJID caused cardiac SODs expressions to be increased.
It can be concluded that CJID prevents H- LVH and VR by reducing ROS production via regulating the NOXs-ROS-NF-ĸB signaling pathway. These findings reveal that CJID protects CVD in hypertension. Further animal and clinical studies are needed to substantiate these important findings.

並列關鍵字

Indonesian herbal medicine ； cardiovascular damage ； oxidative stress ； NADPH oxidases (NOXs) ； nuclear factor kappa B (NF-kB) ； spontaneously hypertensive rats

參考文獻

Ago, T., et al. (2010). "Upregulation of NOX4 by hypertrophic stimuli promotes apoptosis and mitochondrial dysfunction in cardiac myocytes." Circulation Research 106: 1253-1264.

Akasaki, T., et al. (2006). "Increased expression of gp91phox homologues of NAD (P) H oxidase in the aortic media during chronic hypertension: involvement of the renin-angiotensin system." Hypertension Research 29: 813-820.

An, H.-J., et al. (2015). "Isoeugenin, a Novel Nitric Oxide Synthase Inhibitor Isolated from the Rhizomes of Imperata cylindrica." Molecules 20: 21336-21345.

Andarwulan, N., et al. (2010). "Flavonoid content and antioxidant activity of vegetables from Indonesia." Food Chemistry 121: 1231-1235.

Appel, L. J. (1999). "Nonpharmacologic therapies that reduce blood pressure: a fresh perspective." Clinical Cardiology 22: 1-5.