An in-vitro analysis of L-Carnitine mediated rescue of TNF-? induced apoptosis in mice oocytes

Mona Sharma, Ranjana Rana, Ashutosh Halder, Surabhi Gupta
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Abstract

: Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine implicated in various physiological and pathological events. Carnitine is a quaternary amine which plays a significant role in fatty acid oxidation and is reported to produce antiapoptotic effects. Aim of this work was to study the effect of L-Carnitine (LC) on TNF-α induced apoptosis in mice oocytes. : Oocytes were isolated from super ovulated Swiss Albino mice and treated with different concentrations of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100ng/ml) and LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml). TUNEL Assay was done for the biochemical assessment of apoptosis.: Apoptotic indices with different doses of TNF-α (0.1ng/ml, 1ng/ml, 10ng/ml, 100 ng/ml) were 28.5%, 71.4%, 100%, 42.8% respectively. The concentration of TNF-α that produced the highest apoptotic index was 10ng/ml. LC alone in different doses (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1.0 mg/ml) did not elicit any apoptotic signal. Further LC was added in different doses with 10ng/ml TNF-α to study the rate of apoptosis in mice oocytes. Apoptotic index with 10 ng/ml TNF-α and different doses of LC (0.1mg/ml, 0.3mg/ml, 0.5mg/ml, 1 mg/ml) were 25%, 37.5%, 50%, 62.5% respectively. The concentration of LC that reduced the apoptotic index to the maximum was 0.1mg/ml.: Present study could demonstrate the anti-apoptotic effect of LC against apoptotic effects of TNF-α in mice oocytes. The study presents preliminary data suggesting a possible therapeutic role of LC in inflammatory etiologies such as ovarian failure.
左旋肉碱介导的TNF-?诱导小鼠卵母细胞凋亡
肿瘤坏死因子-α (TNF-α)是一种促炎细胞因子,参与多种生理和病理事件。肉碱是一种季胺,在脂肪酸氧化中起重要作用,据报道具有抗细胞凋亡作用。本实验旨在研究左旋肉碱(LC)对TNF-α诱导的小鼠卵母细胞凋亡的影响。:取瑞士白化病超排卵小鼠卵母细胞,分别用不同浓度的TNF-α (0.1ng/ml、1ng/ml、10ng/ml、100ng/ml)和LC (0.1mg/ml、0.3mg/ml、0.5mg/ml、1.0 mg/ml)处理。TUNEL法测定细胞凋亡的生化变化。不同剂量TNF-α (0.1ng/ml、1ng/ml、10ng/ml、100 ng/ml)对小鼠的凋亡指数分别为28.5%、71.4%、100%、42.8%。产生最高凋亡指数的TNF-α浓度为10ng/ml。不同剂量的LC (0.1mg/ml、0.3mg/ml、0.5mg/ml、1.0 mg/ml)均未引起细胞凋亡信号。在不同剂量的LC中加入10ng/ml TNF-α,研究小鼠卵母细胞的凋亡率。10 ng/ml TNF-α和不同剂量LC (0.1mg/ml、0.3mg/ml、0.5mg/ml、1mg/ml)作用下的细胞凋亡指数分别为25%、37.5%、50%、62.5%。使细胞凋亡指数降低最大的LC浓度为0.1mg/ml。本研究可以证明LC对小鼠卵母细胞TNF-α的抗凋亡作用。该研究提供的初步数据表明,LC在卵巢功能衰竭等炎症病因中可能具有治疗作用。
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