土肉桂葉精油對於STZ誘發之糖尿病大鼠發炎體相關蛋白質表現及腎臟病變發展之影響

The effect of leaf essential oil of Cinnamomum osmophloeum Kaneh on the expression of inflammosome related protein and the development of diabetic nephropathy in STZ-diabetes rats

張皓瑜(Hao-Yu Chang)

指導教授 ： 劉承慈

中山醫學大學/健康管理學院/營養學研究所/碩士(2011年)

https://doi.org/10.6834/CSMU.2011.00164

摘要

本研究目的為觀察台灣土肉桂葉精油 (TC)是否可以改善糖尿病大鼠腎病變的發生，同時探討其對糖尿病誘發之NALP3 inflammasome相關分子表現量之影響。實驗使用雄性Wistar大鼠，由尾靜脈注射STZ (65 mg/kgBW) 誘發糖尿病，控制組則由尾靜脈注射不含STZ的生理食鹽水。誘發後三天量測血糖及尿糖，確認是否誘發成功，誘發成功的糖尿病大鼠採隔日灌食，總共灌食八次，分別灌食載劑玉米油 (corn oil,2 ml/kg BW)、低、中、高劑量土肉桂葉精油 (13、26、或54 mg/kg BW)、肉桂醛 (C40；40 mg/kg BW)及口服降血糖藥glibenclamide (GL；1.2 mg/kg BW)，另在控制組灌食載劑玉米油 (corn oil,2ml/kg BW)。，發後第20天收集24小時尿液分析creatinine及albumin的濃度。誘發後第21天以二氧化碳犧牲，收集血清分析creatinine及BUN濃度；收集肝臟、淋巴結、迴腸、腎臟、脾細胞以西方墨點法分析NALP3、ASC、Caspase-1的蛋白質含量；腎臟冷凍切片，以MPO組織化學染色法分析腎臟發炎細胞浸潤程度，同時以HE組織學染色觀察腎臟及腎絲球形態學。結果顯示: 給予不同劑量的TC均可逆轉糖尿病造成的生長代謝異常，效果與C40及GA相當。腎功能指標部分:給與不同劑量的TC均可顯著降低微白蛋白尿及BUN，給予中劑量可顯著降低CCR、GFR，但高劑量只降低CCR，給與C40及GL只顯著降低BUN的含量，對於CCR、GFR及微白蛋白尿則無影響。給予TC、C40及GL後可逆轉糖尿病造成的腎臟發炎細胞浸潤及腎絲球硬化、基底膜及環間質增厚的現象。給予TC在低劑量可以逆轉糖尿病所致之淋巴結跟迴腸ASC的表現量，中劑量可以降低肝臟NALP3跟脾細胞Caspase-1的表現量，而肉桂醛只能有效降低脾細胞ASC的含量。由以上結果可推論，土肉桂葉精油可以有效改善糖尿病大鼠腎臟惡化的情況，避免糖尿病腎病變的產生，同時降低NALP3 inflammasome相關分子的表現且優於肉桂醛，而土肉桂葉精油中活性成分有待進一步釐清。

關鍵字

STZ ； NALP3 ； 發炎體 ； 大鼠 ； 土肉桂 ； 糖尿病腎病變

並列摘要

The aim of the present study is to investigate the effect of leave essential oil of Cinnamomun osmophloeum Kaneh. (TC) on the development of diabetic neuropathy and on the expression of NALP3 inflammasome related molecules. Male Wistar rats were induced to be diabetes with STZ (65mg/kg BW, iv). Control rats were injected with vehicle (citrate buffer, pH 4.2). At three days after injection, rats confirmed to be diabetes were gavaged vehicle (corn oil, 2 ml/kg BW), low-, middle-, or high-dose of TC (12, 26, or 54 mg/kg BW, respectively), cinnamaldehyde (C40; 40 mg/kg BW), or glibenclamide (GL; 1.2 mg/kg BW) every other day and for eight times. Control rats were gavaged corn oil (2 ml/kg BW). On day 20 after the induction of diabetes urine were collected during a 24 h period for the analysis of creatinine and albumin. Rats were killed followed by the collection of blood for the analysis of creatinine and BUN and organs and tissues were collected for Western blotting analysis on NALP3, ASC, and Caspase-1. The cryostat sections of kidney were prepared for MPO staining and cross-sections of paraffin-embedded kidney were prepared for morphological study. It was found that all tested doses of TC reversed diabetes-induced metabolic changes to an extent similar to that by C40 and GL. All tested doses of TC also significantly reversed micro-albumin urine and BUN in diabetes; middle-dose of TC significantly ameliorated CCR and GFR while high-dose of TC only ameliorated CCR. Both C40 and GL ameliorated BUN but neither of them affected CCR, GFR, or micro-albumin urine. All doses of TC, C40 and GL ameliorated phagocyte infiltration and normalized morphology of kidney in diabetes.Low-dose of TC reversed elevated ASC in lymph nodes and ileum, middle-dose of TC reversed NALP3 in liver and Caspase-1 in splenocytes, while C40 only reversed ASC in splenocytes in diabetes. In conclusion, the present study showed a prophylactic effect of TC on the development of diabetes nephropathy and lowered the expression of NALP3 inflammasome related molecules induced by diabetes and such effects of TC is greater than cinnamaldehyde. The functional composition in TC remained to be clarified in the future.

並列關鍵字

STZ ； NALP3 inflammasome ； Rats ； Cinnamomun osmophloeum Kaneh ； Diabetic neuropathy

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