補充鱸魚精對於cisplatin誘發之副作用的保護效果及機制

The protective effects and the mechanisms of perch essence on cisplatin-induced side effects

游舒涵(Shu-Han Yu)

指導教授 ： 葉姝蘭

中山醫學大學/健康管理學院/營養學系碩士班/碩士(2024年)

摘要

Cisplatin (cis-diamminedichloroplatinum (II), CDDP )是臨床上廣泛使用治療癌症的一線化療藥物，但因其對除了癌細胞以外的正常器官組織具有無差別毒性，除腫瘤外亦會對許多組織產生毒性，降低病患生活品質，而限制了cisplatin的使用，故有必要尋找有效的策略改善其副作用。近年來許多研究顯示，魚類的蛋白水解物或胜肽對人體健康可能有幫助，鱸魚精（perch essence, PE）是以鱸魚之魚頭、魚骨、魚肉及魚皮經水在高溫高壓條件下萃取而得的產品，其富含小分子胜肽及支鏈氨基酸。有動物研究顯示PE能降低運動引發的疲憊，但對於化療藥物cisplatin誘發的副作用尚缺乏相關研究。因此，本研究進行了兩項動物研究，探討鱸魚精對 cisplatin 誘發的疲勞、肌肉流失、睪丸損傷和骨髓抑制的保護效果及相關機制。我們首先進行了為期2週的短期研究，將雄性BALB/c小鼠隨機分為五組: Control、CDDP (7mg/kg/week, i.p)、CDDP+4XPE（4倍劑量鱸魚精）、CDDP+2XPE （2倍劑量鱸魚精）、CDDP+T （與4XPE相同蛋白量之tryptone作為對照組）。接下來，我們進行了八週的長期研究，將雄性BALB/c小鼠隨機分為五組: Control、CDDP (2.5mg/kg/week, i.p)、CDDP+2XPE （2倍劑量鱸魚精）、CDDP+1XPE (1倍劑量鱸魚精)、CDDP+T （與2XPE相同蛋白量之tryptone作為對照組）。整體而言，這兩項研究結果一致顯示，補充鱸魚精有減少cisplatin引起的疲勞、肌肉流失、睪丸損傷和骨髓抑制的趨勢。然而，在長期研究中，我們發現鱸魚精的改善效果優於短期研究，且在兩項實驗當中2XPE皆具有更好的保護效果，且效果比tryptone好，因此我們推測2倍劑量為鱸魚精較佳的濃度。長期研究的主要結果顯示：1. 與 CDDP 組相比，鱸魚精增加了小鼠抓力及活動力，並且減少了肌肉的流失，同時增加了肌肉纖維橫截面積(cross-sectional area, CSA)、myosin heavy chain (MyHC)表現及肝醣含量。進一步研究發現，鱸魚精可以調控肌肉分解相關蛋白MuRF-1、Atrogin-1的表現。2.鱸魚精可以緩和cisplatin誘導的精子濃度和活動力下降、睪丸萎縮等睪丸毒性，這些機制與降低細胞凋亡相關蛋白(Bax、caspase-3、cleaved caspase-3、caspase-9、cleaved caspase-9)並增加PCNA蛋白表現量有關。3.鱸魚精還能改善cisplatin誘導的骨髓抑制，因鱸魚精能回升骨髓細胞數量，而鱸魚精的作用與骨髓細胞中TNF-α濃度降低和IL-9濃度提高有關。4.上述鱸魚精的效果與降低脂質過氧化程度(2-thiobarbituric acid reacting substances test, TBARs)和促發炎激素TNF-α、IL-6或IL-1β 有關。

關鍵字

Cisplatin ； 鱸魚精 ； 睪丸毒性 ； 骨髓抑制 ； 肌肉流失

並列摘要

Cisplatin (CDDP) is a widely used first-line chemotherapeutic agent for cancer treatment. However, due to its nonselective cellular toxicity, cisplatin induces various side effects that reduce the quality of life of the patients. Therefore, it is necessary to find effective strategies to improve the side effects of cisplatin. In recent years, studies have shown that the extracts or peptides of fish may benefit to human health. Perch Essence (PE), an extract from the head, bones, flesh, and skin of perch, is rich in small molecule peptides and branched-chain amino acids and has been shown to reduce exercise-induced fatigue in animal studies. However, whether PE improves the side effects of cisplatin remains unclear. Therefore, in the present study, we conducted two animal studies to investigate the protective effect of PE against cisplatin-induced fatigue, muscle loss, testis damage, and myelosuppression as well as the possible mechanisms. We first performed a short-term study (2 weeks), in which male BALB/c mice were randomly assigned into five groups: Control, CDDP (7mg/kg/week, i.p.), CDDP+4XPE (quadruple dose of PE/packet), CDDP+2XPE (double dose of PE/packet), and CDDP+T (tryptone with the same level of protein as 4XPE). Next, we performed a long-term study (8 weeks), in which male BALB/c mice were randomly divided into the following groups: Control, CDDP (2.5 mg/kg/week, i.p.), CDDP+2XPE (double dose of PE/packet), CDDP+1XPE (single dose of PE/packet), CDDP+T (tryptone with the same level protein as 2XPE). In general, the results of these two studies consistently showed that supplementation with PE tended to attenuate fatigue (evidenced by increasing grip strength and locomotor activity), muscle loss, testis damage, and myelosuppression induced by CDDP. However, the efficacy of PE supplements was better in the long-term study than that in the short-term study. 2XPE had a better protective effect in both studies than T (with an equivalent protein level as 2XPE or 4XPE). In long-term study, the major findings were 1. PE increased grip strength and locomotor activity accompanied by decreasing muscle weight loss and increasing muscle fiber cross-sectional area (CSA), MyHC expression, and muscle glycogen in mice exposed to CDDP. Further studies revealed that PE decreased the expression of muscle catabolism-related proteins, MuRF-1 and Atrogin-1. 2. PE mitigated cisplatin-induced testicular toxicity by increasing sperm concentration and motility, and decreasing testicular atrophy. The mechanisms were associated with reducing apoptosis-associated proteins (Bax, caspase-3, cleaved caspase-3, caspase9, cleaved caspase-9) and increasing PCNA protein expression. 3. PE also improved cisplatin-induced myelosuppression because PE supplements increased bone marrow cell count. The effect of PE was associated with a reduction of TNF-alpha levels and an increase of IL-9 levels in bone marrow cells. 4. The effects of the PE mentioned above were associated with its antioxidant and anti-inflammatory effects.

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