大蒜精油對糖尿病大鼠心臟病變的影響評估與精神分裂症病人單核球輔助型T細胞激素及TLR訊息RNA表現之分析

The Assessment of Impact about Garlic Oil on Cardiomyopathy in Diabetic Rats and Analysis of T cell Cytokines and TLR mRNA Expression in Patients with Schizophrenia

張勝皇(Sheng-Huang Chang)

指導教授 ： 曾博修

中山醫學大學/醫學院/生化暨生物科技研究所/博士(2014年)

https://doi.org/10.6834/CSMU.2014.00120

摘要

PART Ⅰ
糖尿病(Diabetes Mellitus, DM)是一種慢性的代謝異常疾病，導因於患者無法有效利用血糖，使體內血糖過高，長期誘發相關併發症，影響病人存活。目前已有相當多文獻指出大蒜精油可以有效控制血糖、促進血液纖維蛋白分解、降血脂、降血膽固醇濃度、抗氧化等進而保護心血管系統，然而保護機制尚未明朗，本研究於是以罹患糖尿病大白鼠動物實驗模式來探討大蒜精油是否可以改善心臟細胞肥大和心肌纖維化的發生。
我們採用由STZ誘導糖尿病的大白鼠，經過確認尿糖及血糖的濃度，觀察誘發糖尿病成功的大白鼠，每2天餵食大蒜精油一次，共餵食16天(Control、0mg/kgBw、10mg/kgBw、500mg/kgBw、100mg/kgBw)後，心臟之胚胎基因(ANP、BNP)和心臟肥大訊息路徑(ERK、JNK、P38、IL-6)以及心肌纖維化訊息路徑(uPA、PAI-1)相關的蛋白表現。
結果得知，心臟肥大及心肌纖維化相關蛋白的表現，因處理大蒜精油而下降，改善糖尿病引起的心臟肥大和心肌纖維化，且呈現dose-dependent的情形。心臟超音波的檢查結果，也顯示大蒜精油也具有改善心臟收縮之功能。由此可知，大蒜精油透過抑制MAPKs (ERK、JNK、P38)、IL-6與uPA、PAI-1之表現，改善糖尿病引起的心臟肥大和心肌纖維化現象。
PART Ⅱ
目前對於細胞激素在於精神分裂症的免疫調節機制及致病機轉之關聯瞭解仍然有限。因此本計畫目的是為瞭解調控體內免疫環境之輔助型T細胞激素（T helper cell cytokine）與Toll-like receptors（TLRs）表現狀況及關聯性，在精神分裂症病人與正常人之差異，從中找出可以作為診斷及判別指標之參考。進而應用到其他疾病的治療或預防。
收集經診斷罹患精神分裂症且常規服用抗經神病藥物的男性病人（n=46；平均年齡40±10.8）與沒有服用藥物的正常男性（n=22；平均年齡33±5.9）之周邊血液與血球進行TLRs與細胞激素基因表現的變化。
本研究發現精神分裂症病人單核球中，其IL-4及IL-10的mRNA表現比率，較正常人單核球的表現比率高，而TLR-3及TLR-5的表現比率，較正常人單核球的表現比率低。顯然精神分裂症病人，其免疫調節基因轉錄層次確與正常人不同。
精神分裂症病人單核球之IL-4與IL-10之mRNA比起正常人有較高之表現率，加上較低的TLR-3及TLR-5之mRNA的表現率。這是首次有這樣的發現，或許可作為精神分裂症病人遺傳的線索。相信對於精神分裂症之診斷及癒後之追蹤，可以提供有效之輔助生物標記。另外，精神分裂症是一種慢性疾病，致病的成因雖不清楚，目前仍無法根治，但是由IL-4、IL-10蛋白表現與正常人相仿的實驗結果，我們認為與精神分裂症住院病人接受持續之醫療照護，固定接受抗精神病藥物之治療有關。

關鍵字

大蒜 ； 心臟 ； 糖尿病

並列摘要

PART Ⅰ
Diabetes is a group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. The diabetic patient can not use glucose effectively with hyperglycemia which will induce related comorbidity that will effect the patient’s survival.
Garlic oil has been reported to promote fibrinogenolysis, reduce cholesterol, inhibits the oxidation and protect the cardiovascular system; however, the effects and mechanisms behind the cardioprotection of garlic oil on diabetes-induced cardiomyopathy (e.g., cardiac hypertrophy and fibrosis ) are unclear. We used streptozotocin (STZ)-induced diabetic rats to investigate whether garlic oil could protect the heart from diabetes-induced cardiomyopathy. Wistar STZ-induced diabetic rats received garlic oil (0, 10, 50 or 100 mg kg−1 body weight) by gastric gavage every 2 days for 16 days. Normal rats without diabetes were used as control.
Then cardiac embryo proteins(atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP)), cardiac fibrosis by uPA and PAI-1, signaling pathway for cardiac hypertrophy-related mitogen-activated protein kinases (MAPK) pathways (e.g., p38, c-Jun N-terminal kinases (JNK) and extracellular responsive kinase (ERK1/2)) , the IL-6/MEK5/ERK5 signaling pathway related protein were observed.
The results revealed that cardiac hypertrophy and fibrosis related proteins were reduced after garlic oil treatment, and were improve in dose-dependent variable. The echocardiography showed garlic oil could better the contraction cardiac contractile function. The garlic oil could suppress MAPKs(ERK1/2, p38, JNK), IL-6 and uPA, PAI-1 representation to alleviate cardiac hypertrophy and fibrosis caused by diabetes mellitus of rats.
PART Ⅱ
There is limited understanding about immune regulation mechanism and etiology of schizophrenic patients. In the study, we tried to understand the presentation and relationship between immune regulatory T helper cell cytokines and Toll-like receptors（TLRs）in schizophrenic patients and normal controls. We hope we can find some indicators to aid on diagnosis, treatment and prevention of schizophrenia and other diseases.
46 schizophrenic male patients（average age 40±10.8）who were using antipsychotic medication regularly and 22 drug-naïve normal males（average age 33±5.9）were enrolled in this study . we collected their peripheral blood samples and blood cells for cytokine and TLRs presentation analysis.
In our study, we found the mRNA presentation rate in IL-4 and IL-10 in monocytes are higher, but TLR-3 and TLR-5 are lower in schizophrenic patients than in normal control. Obviously the immune regulation gene transcriptions were different between schizophrenic patients and normal controls.
The presentation rate in mRNA of IL-4and IL-10 are higher, but TLR-3 and TLR-5 are lower in monocytes among schizophrenic patients. This finding may be used in genetic counseling for diagnosis of schizophrenic patients as a useful biological maker. In addition, after continuous antipsychotic treatment, the IL-4 and IL-10 protein presentation were similar between schizophrenic patients and normal controls. So it is important to push schizophrenic patients to receive continuous treatments and relapse prevention for the sake of their safty and health.

並列關鍵字

Garlic oil ； Cardiomyophthy ； Diabetes

參考文獻

1. American Diabetes Association (2013) Diagnosis and classification of diabetes mellitus. Diabetes care 36: S67–74

2. Marian AJ, Roberts R (1995) Recent advances in the molecular genetics of hypertrophic cardiomyopathy. Circulation 92 : 36–1347

3. Marian, A.J., Roberts, R. (1995) Recent advances in the molecular genetics of hypertrophic cardiomyopathy. Circulation, 92 , 36–1347

4. Boheler, K.R., Schwartz, K. (1992) Gene expression in cardiac hypertrophy. Trends Cardiovasc Med., 5 ,176–182

5. Boheler KR, Schwartz K (1992) Gene expression in cardiac hypertrophy. Trends Cardiovasc Med 5 : 176–182