- 學位論文
觀察柴胡及小柴胡湯濃縮藥粉對第二型糖尿病小鼠的保腎作用
Renal protect effect of Radix Bupleuri and Xiao-Chai-Hu-Tang concentrated powders on type 2 diabetic mice.
摘要
動機：糖尿病相關腎疾病的盛行率逐年增加，成為日益嚴重的健康議題及全民健保財務負擔。據文獻報導柴胡皂苷B2可減輕腎纖維化作用，小柴胡湯水萃物可改善第一型糖尿病腎病模型小鼠的腎功能。 目的：觀察臨床使用的柴胡及小柴胡湯濃縮藥粉對第二型糖尿病小鼠是否具有保腎能力。 方法：以C57BL/6雄性小鼠經腹腔靜脈注射低劑量Streptozotocin (45 mg/kg/day)合併高脂飼料餵養建立第二型糖尿病小鼠模型。將此糖尿病小鼠分成六組(n=8)：實驗組分別為高低劑量柴胡組(133 mg或66.5 mg柴胡濃縮藥粉/kg)，以及高低劑量小柴胡湯組(131 mg或65.5 mg小柴胡湯濃縮藥粉/kg)；糖尿病對照組給予等量載體（生理食鹽水），正對照組給予Metformin (250 mg/kg)，一週5次，連續給予15週。另保留同一批小鼠8隻餵以正常飼料作為正常對照組，每日給予等量載體。每5週取得小鼠的血糖值、糖化血色素(HbA1c)、血清尿素氮(BUN)及血清肌酸酐(Serum creatinine, Scr)。治療15週後犧牲小鼠，收集尿液及血清樣本冷凍保存在-80°C備用。取右腎包埋固定進行組織切片染色檢測；將左腎稱重並冷凍以供將來研究。 結果：柴胡組及小柴胡湯組在第10週時的BUN及Scr數據相比於糖尿病對照組有明顯差異(p<0.05)，但在第15週時無明顯差異。腎指數 (左腎重/體重)相比於糖尿病對照組，低劑量柴胡組表現出極顯著差異(p<0.01)，表示腎水腫的狀況最輕微。單次尿蛋白排出量相比於糖尿病對照組是柴胡組最佳（高劑量僅49.3 %，低劑量僅45.8 %）。腎臟切片HE染色可看出糖尿病小鼠各組都有些許腎絲球系膜增生及腎小管擴張現象；Masson染色下看到實驗各組相較起糖尿病對照組的腎臟切片在腎小管間質組織有較少程度的膠原纖維顯色。 結論：柴胡及小柴胡湯濃縮藥粉具有改善第二型糖尿病小鼠腎功能的療效。
並列摘要
Background: The increasing prevalence rate of diabetic kidney disease is a crucial health issue and heavy burden of National Health Insurance. A few researches revealed that Saikosaponin B2, a biologically active ingredients of Radix Bupleuri, and Xiao-Chai-Hu -Tang have the potential ability to improve renal function and reduce renal fibrosis in DKD. Objective: To observe if Radix Bupleuri and Xiao-Chai-Hu-Tang concentrated powders have renal protect effect on type 2 DM mice. Methods and Materials: Build type 2 DM model by using C57BL/6 male mice to undergo streptozotocin injection and high fat diet. Type 2 DM mice were randomly divided into 6 groups(n=8) and received Radix Bupleuri(133 mg or 66.5 mg concentrated powder/kg), Xiao-Chai-Hu-Tang(133 mg or 66.5 mg concentrated powder/kg), Metformin (250 mg/kg) or vehicle for 15 weeks. The other 8 mice were fed general diet and injected normal saline do be normal control group. Get body weights, blood glucose, HbA1C, BUN and serum creatinine data of mice every 5 weeks. Sacrifice the mice after oral gavage for 15 weeks and collect urine and blood samples, weight left kidneys. Store these samples in -80°C refrigerator. Fix right kidneys in formalin solution and then embed these in paraffin for observing histological morphology changes. Results: Comparing with DM control group, BUN Scr at 10th weeks of Radix Bupleuri and Xiao-Chai-Hu-Tang groups showed significant differences ( t-test, p<0.05). Kidney index (left renal weight over body weight) of Radix Bupleuri group showed significant difference (p<0.01) and the urine albumin excretion amounts were approximately half of the DM control group. Kidney slides with HE stain showed glomerular mesangial expansion and tubular dilation. Masson's trichrome stain slides of Radix Bupleuri and Xiao-Chai-Hu groups looked less collagen fibers in renal interstitial. Conclusion: Radix Bupleuri and Xiao-Chai-Hu-Tang concentrated powders have renal protect ability to type 2 DM mice.