- 學位論文
蘆薈複方應用於改善腺嘌呤導致小鼠慢性腎臟疾病之研究
The protective effect of aloe vera complex formula on mice from adenine-induced chronic kidney disease
摘要
腺嘌呤(adenine)在體內的代謝產物2,8-二羥基腺嘌呤(2,8-dihydroxy adenine, 2,8-DHA)，會沉積於腎臟引起纖維化(fibrosis)，最終導致慢性腎臟疾病(chronic kidney disease, CKD)。蘆薈萃取物(aloe vera extract, AV)已被證實具有抗氧化、抗發炎、抑制細菌活性及促進傷口癒合的功效。在中醫學的觀點，認為豬腎(pig kidney, PK)可能具有改善腎功能的作用。因此，本研究將探討AV複方後對於腎臟保護的作用及分子機制。十週實驗期間，小鼠先給予腺嘌呤處理，再餵食含有AV (1%和2%)和PK (1%)單獨或合併方式。結果顯示，餵食AV複方能夠顯著增加腎臟/體重比值以及減少腺嘌呤誘導的腎臟纖維化。H＆E染色中也發現腎小管構造較為完整且密集。在餵食AV複方後能夠降低血清尿素氮(blood urea nitrogen, BUN)和肌酸酐(creatinine, CRE)含量。此外，腺嘌呤處理的小鼠腎臟組織，在AV複方處理能夠提升總抗氧化能力(TEAC)與抗氧化酵素過氧化氫酶(catalase)、穀胱甘肽(glutathione, GSH)、穀胱甘肽過氧化酶(glutathione peroxidase, GPx)、超氧化物歧化酶(superoxide dismutase, SOD)和穀胱甘肽還原酶(glutathione reductase, GRd)的活性以及維持較高的GSH含量。腎臟脂質過氧化程度(TBARs)與促發炎因子(IL-6和TNF-α)之濃度，在餵食AV複合PK後也進一步降低。分子數據顯示，AV複方之抗纖維化作用，可能經由調控TGF-β1/Smad訊息傳遞路徑，進而減少纖維化蛋白(α-SMA、fibronectin、PAI-1、type 1 collage和CTGF)。同時AV複方處理也透過減少發炎因子(NF-κB、iNOS、和COX-2)降低腎臟發炎反應。總結，AV複方不僅具有抗氧化、抗發炎作用，亦能調節TGF-β1/Smad訊息傳遞路徑，保護腎臟免於腺嘌呤誘導CKD進展。
並列摘要
Adenine metabolites, 2,8-DHA, in vivo deposited in the kidney tissues cause fibrosis, eventually to chronic kidney disease(CKD). Aloe vera extract (AV) has been shown to posse antioxidant, anti-inflammatory, and bacteriostatic activities, and promote wound-healing. In the view of Chinese medicine, pig kidney (PK) seems to improve kidney function. Here, we investigated the kidney-protective effects and the molecular mechanism(s) of AV combined PK in vivo. First, the mice were treated adenine supplemented with AV (1% and 2%) or/and PK (1%) during the 10-week treatment period. Result showed that feeding AV complex to mice significantly increased the ratio of kidney weight to body weight and reduced the characteristic of renal fibrosis induced by adenine treatment. H＆E data showed that tubular structure of kidney is more complete and dense. The serum levels of BUN and CRE also were decreased by the AV complex. Additionally, the kidney tissues of adenine-treated mice, AV complex increased the activities of total antioxidant capacity (TEAC) and antioxidant enzymes catalase, glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD), and glutathione ruductase (GRd) and maintain a high GSH content. The thiobarbituric acid relative substances (TBARs) and pro-inflammatory cytokines (IL-6 and TNF-α), further reduced by the AV combined PK. Molecular data showed the anti-fibrotic effect of AV complex might be mediated via TGF-β1/Smad signaling pathway, which in turn led to reduced the levels of fibrosis factors (α-SMA, fibronectin, PAI-1, type 1 collage, and CTGF) in kidney tissues. AV complex also reduced protein levels of the inflammatory factors (NF-κB, iNOS, and COX-2). Our data imply that AV complex possesses antioxidant, and anti-inflammative, and TGF-β1/Smad modulation anti-fibrotic effect, which in turn led to protect the kidney from adenine-induced CKD.