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Title: | 藉由蘭嶼豬探討益生菌減緩順鉑誘發慢性腎衰竭之研究 Effect of Potential Probiotics on Alleviating Chronic Kidney Disease Progression Using Cisplatin-Induced Lanyu Porcine Model |
Authors: | Pin-Jhu Wang 王品筑 |
Advisor: | 陳明汝 |
Keyword: | 慢性腎臟病,益生菌,蘭嶼豬,腸道菌相,硫酸??酚, Chronic kidney disease,Probiotic,Lanyu pig,Intestinal microbiota,Indoxyl sulfate, |
Publication Year : | 2016 |
Degree: | 碩士 |
Abstract: | 慢性腎臟病為全球共同關注的健康議題,美國約有2600萬位成年人得到不同程度的慢性腎臟病。流行病學研究指出,糖尿病病患和高血壓病患也是罹患慢性腎臟病的高風險族群。通常慢性腎臟病是緩慢且漸進式地喪失腎臟功能,當腎功能損失程度達到後期腎臟病變時,會伴隨著尿毒症的發生。治療方式為透過血液透析減少累積在體內的尿毒素,然而血液透析無法有效地清除與蛋白質小分子結合的尿毒素像是硫酸吲哚酚(indoxyl sulfate, IS)。累積在體內的硫酸吲哚酚進而影響腎臟之功能,造成慢性腎臟病之惡化,研究指出硫酸吲哚酚與慢性腎臟病病程階段以及死亡率有關聯性。
實驗室先前研究證實了將Lactobacillus plantarum subsp. plantarum、Lb. paracasei subsp. paracasei和Streptococcus salivarius subsp. thermophilus三株菌以等比例方式混合稱之為混菌Pm-1,在體外試驗中,混菌Pm-1對於培養基內硫酸吲哚酚之清除效果比起其他單一菌株更好。以順鉑(cisplatin)誘導急性腎衰竭之大鼠模式中,混菌Pm-1有效地降低了大鼠血液中硫酸吲哚酚含量。 由於目前尚未建立慢性腎臟病豬隻模式,本次實驗目的為建立以cisplatin誘導慢性腎臟病蘭嶼豬模式,探討給予混菌Pm-1於cisplatin誘導慢性腎臟病之影響。試驗設計共分成4組,每組3隻8月齡蘭嶼豬。組別分成控制組,給予一般飼糧;cisplatin誘導組,一般飼糧且以靜脈注射cisplatin;低劑量Pm-1組,給予109 CFU/kg BW Pm-1且以靜脈注射cisplatin;高劑量Pm-1組,給予1010 CFU/kg BW Pm-1且以靜脈注射cisplatin,混菌Pm-1會在cisplatin誘導前先給予90天。 結果顯示,高劑量Pm-1組別其血液中肌酸酐及尿素氮之含量比起cisplatin 誘導組有下降的趨勢。透過蘇木紫─伊紅染色和Masson's trichrome染色,高劑量Pm-1組腎臟組織病灶發生率及腎臟纖維化程度皆低於cisplatin誘導組。腎臟組織使用化學免疫組織染色法偵測凋亡蛋白酶-3之表現量,高劑量Pm-1組凋亡蛋白酶-3面積比起cisplatin誘導組有下降的趨勢。高劑量Pm-1組可以降低蘭嶼豬血液中tumor necrosis factor (TNF)-α和interleukin (IL)-6以及過氧化氫酶之含量。在糞便菌相部分,高劑量Pm-1組比起cisplatin誘導組lactobacilli和bifidobacteria有增加,而Clostridium perfringens則是下降。然而給予Pm-1之組別,其血液中硫酸吲哚酚含量並沒有顯著差異。在腸道菌相部分,由熱圖以及主成分分析圖觀察到給予高劑量Pm-1作為前處理,其腸道菌相組成比起cisplatin誘導組更接近控制組。 綜觀上述,給予高劑量Pm-1於cisplatin誘導慢性腎臟病蘭嶼豬在預防其腎臟受損之能力有很好的效果。 Chronic kidney disease (CKD) is a global health issue, approximately 26 million American adults are under CKD status. Epidemiological evidences revealed that the patient with diabetes and high blood pressure accompany with high CKD risk. In general, CKD characterized by a gradual dysfunction of kidney, and then caused uremia syndrome. Hemodialysis (HD) is the most common method used to treat uremia syndrome, however, HD is unable to effectively eliminate protein-bound solutes in the advanced stage of CKD. Thus, the accumulation of protein-bound uremic toxins, including indoxyl sulfate (IS), has been suggested to be related to complications and mortality in HD patients. Our previous study clearly demonstrated that the combination of three strains, Pm-1 which includes Lactobacillus plantarum subsp. plantarum, Lb. paracasei subsp. paracasei and Streptococcus salivarius subsp. thermophilus, have the better ability to reduce uremic toxin, IS, in MRS broth under in vitro and in rat models. In the present study, we developed a cisplatin-induced CKD model using Lanyu pig and investigated the effects of Pm-1 on preventing CKD using this model. Eight-month-old Lanyu pig were divided into four groups, containing three pigs in each group. Nephrotoxicity was induced by intravenous (i. v.) injection of cisplatin dissolved in normal saline. The groups were divided into Control (normal diet without cisplatin injection), CP (normal diet with cisplatin injection), Low Pm-1+CP (1×109 CFU/kg BW Pm-1 with cisplatin injection) and High Pm-1+CP group (1×1010 CFU/kg BW Pm-1 with cisplatin injection). The results indicated that the pigs fed with high dosage of Pm-1 showed the trend to reduce both creatinine and blood urea nitrogen (BUN) when compared with cisplatin group. The high dosage Pm-1 group also demonstrated the lower incidence of lesions in H&E stain and lower fibrosis area in Masson's trichrome stain. The high dosage Pm-1 group also show the lower cleaved caspase-3 positive area on kidney cell apoptosis compared with cisplatin group accompany with suppression of tumor necrosis factor (TNF)-α and interleukin (IL)-6 production and elevation of catalase activity in plasma. In fecal microbiota, Pm-1 administration group had more lactobacilli and bifidobacteria proportion while less Clostridium perfringens when compared with cisplatin group. High dosage Pm-1 show the similar intestinal microbiota component compare with control group in heatmap and principal component analysis (PCA) of the abundance of intestinal microbiota. According to the above results, administration of high dose Pm-1 as pretreatment could alleviate cisplatin-induced CKD in a porcine model. The impressive impact of pretreatment with Pm-1 on the composition of the gut microbiome in cisplatin-induced CKD Lanyu pig model was evidenced by the differences in relative microbial abundance. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78139 |
DOI: | 10.6342/NTU201602356 |
Fulltext Rights: | 有償授權 |
metadata.dc.date.embargo-lift: | 2026-08-11 |
Appears in Collections: | 動物科學技術學系 |
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