桂花、菊花甲醇萃物對小鼠腎間質細胞表現細胞外基質蛋白與脂質堆積之影響

Effects of Osmanthus fragrans, Chrysanthemum morifolium methanolic extracts on the expression of extracellular matrix proteins and lipid accumulation in mouse renal mesangial cells.

莊仲潔(Chung-Chieh Chuang)

指導教授 ： 吳文惠

國立臺灣師範大學/教育學院/人類發展與家庭學系/碩士(2011年)

摘要

台灣的末期腎病的發生率與盛行率在世界44國中居第一。最近流病研究發現單是腹部肥胖就可引起微白蛋白尿與慢性腎病的組織病理特徵，其發生可能早於高血壓與高血糖。腹部肥胖、第二型糖尿病、脂質代謝異常與胰島素阻抗者，其血中含有高濃度游離脂肪酸，促使腎間質細胞堆積脂質，過度分泌基質蛋白如collagen及fibronectin等，與慢性腎病腎絲球纖維化有極大相關性。
本實驗以小鼠腎間質細胞 (Mouse mesangial cell; MC)為模式，以200 μM oleate誘發腎間質細胞堆積脂質、分泌促纖維化生長因子transforming growth factor-β(TGF-β)，與誘發細胞基質蛋白過度生成，並探討數種植物萃取物的影響。結果發現，茉莉、菊花、桂花甲醇萃物，丁香、迷迭香、紫蘇、花椒、鼠尾草、百里香、洋香菜、茴香乙醇萃物，以及苜蓿乙酸乙脂萃物可顯著抑制腎間質細胞TGF-β的分泌，而甘草乙醇萃物、山苦瓜乙酸乙酯萃物則無顯著影響。選擇可最有效抑制TGF-β分泌的桂花、菊花甲醇萃物做進一步研究，發現桂花甲醇萃物可降低細胞內三酸甘油酯及膽固醇含量，菊花甲醇萃物在高劑量時可降低細胞內三酸甘油酯及膽固醇含量，兩者均可顯著抑制腎間質細胞fibronectin mRNA表現並抑制fibronectin過度分泌。另外桂花甲醇萃物可抑制oleate所誘發之reactive oxygen species (ROS)生成，而菊花甲醇萃物可抑制oleate所誘發之extracellular signal-regulated kinase (ERK)磷酸化。
以上結果顯示桂花、菊花甲醇萃物可降低高游離脂肪酸所誘發腎間質促纖維化生長因子分泌、脂質堆積與基質蛋白増生，對於腹部肥胖所導致的腎絲球纖維化可能有預防效果。

關鍵字

腎間質細胞 ； 桂花 ； 菊花 ； 脂質堆積

並列摘要

Recent epidemiological studies show central obesity can induce microalbuminuria and pathological features of chronic kidney disease which may appear before hypertention and hyperglycemia. In obesity, type II diabetes, hyperlipidemia and conditions of insulin resistance, the concentration of circulating free fatty acid increases, and is associated with increasing lipid accumulation and extracellular matrix protein secretion (e.g collagen, fibronectin) in mesangial cells, which may result in glomerular fibrosis.
In this study, mouse mesangial cells (MCs) were treated with 200 μM oleate to increase the accumulation of lipids, the secretion of transforming growth factor (TGF)-β, and the expression of fibronectin. The protective effects of several herbal extracts were tested. We found the methanolic extracts of Jasmine, Chrysanthemu and Osmanthus, the ethanolic extracts of Clove, Perilla, Zanthoxylum, Salvia, Thymus, Parsle and Fennel, and the ethyl acetate extracts of Rosemary and Alfalfa significantly inhibited secretion of TGF-β induced by oleate in mouse MCs, but Glycyrrhiza ethanolic extract and bitter gourd ethyl acetate extract had no significant effects. Among them, Osmanthus fragrans (OF) and Chrysanthemum morifolium (CM) methanolic extracts, especially OF having the best TGF-β inhibition effects were investigated further. We observed that OF methanolic extract at concentration of 50-200 μg/mL decreased intracellular triacylglycerole and cholesterol levels, so did CM methanolic extract at high concentration (200 μg/mL) Both OF and CM methanolic extracts suppressed oleate-induced fibronectin mRNA overexpression, and fibronectin secretion in MCs. We also observed that OF methanolic extract decreased oleate-induced ROS production, and CM methanolic extract suppressed oleate-induced ERK phosphorylation.
In conclusion, OF and CM methanolic extracts prevent oleate-induced fibrosis factor, TGF-β, and ECM oversecretion, and decrease intracellular lipid accumulation. OF and CM methanolic extracts may have protective effects on high plasma free fatty acid-induced chronic kidney disease.

並列關鍵字

mesangial cell ； Osmanthus fragrans ； Chrysanthemum morifolium ； lipid accumulation ； fibronectin

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