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簇分化抗原 147(CD147)的表达与萎锈灵诱导的软骨细胞凋亡有关,这种凋亡是在绿原酸反应下,通过 Bcl-2/Bax/Caspase-3 信号通路发生在胫骨生长板中的。

Cluster of differentiation 147 (CD147) expression is linked with thiram induced chondrocyte's apoptosis via Bcl-2/Bax/Caspase-3 signalling in tibial growth plate under chlorogenic acid repercussion.

机构信息

College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.

College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China; Institute of Traditional Chinese Veterinary Medicine, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Ecotoxicol Environ Saf. 2021 Apr 15;213:112059. doi: 10.1016/j.ecoenv.2021.112059. Epub 2021 Feb 27.

Abstract

Tibial dyschondroplasia (TD) is a metabolic disease of young poultry that affects bone andcartilage's growth. It mostly occurs in broilers due to thiram toxicity in the feed. In this disease, tibial cartilage is not yet ripe for ossification, but it also results in lameness, death, and moral convictions of commercial poultry due to numerous apoptotic changes on cell level. These changes serve a cardinal role in this situation. Many potential problems indicate that chlorogenic acid (CGA) performs an extensive role in controlling apoptosis's perception. However, the actual role of CGA in TD affected chondrocytes in-vitro is still unidentified. The current study investigates the imperceptible insight of CGA on chondrocyte's apoptosis via B-cell lymphoma 2 (Bcl-2), Bcl-2 associated x-protein (Bax), and Caspase-3 with CD147 signalling. The expression of these markers was investigated by Immunofluorescence, western blot analysis, and reverse transcription-quantitative polymerase chain (RT-qPCR). Chondrocytes from the growth plate of tibia were isolated, cultured, and processed. A sub-lethal thiram (2.5 μg/mL) was used to induce cytotoxicity and then treated with an optimum dose (40 μg/ mL) of CGA. According to the results, thiram distorted chondrocyte cells with enhanced apoptotic rate. But, in case of CGA, high expression of CD147 enhanced cell viability of chondrocytes, accompanied by downregulation of Bax/Caspase-3 signalling with the upregulation of Bcl-2. The first possibility has ruled out in the present study by the observation that the cells apoptosis marker, Caspase-3 showed a significant change in CD147 overexpressing cells. Conversely, immunodepletion of CD147 with enhanced cleavage of Caspase-3, indicating the activation of apoptosis in chondrocytes cells. Therefore, these findings suggest a novel insight about CD147 in thiram induced TD about the regulation of Bcl-2/Bax/Caspase-3 apoptosis-signalling axis.

摘要

胫骨软骨发育不良(TD)是一种影响骨骼和软骨生长的幼禽代谢疾病。由于饲料中的 thiram 毒性,它主要发生在肉鸡中。在这种疾病中,胫骨软骨尚未成熟进行骨化,但由于细胞水平上大量的细胞凋亡变化,也会导致跛行、死亡和商业家禽的道德信念。这些变化在这种情况下起着至关重要的作用。许多潜在的问题表明,绿原酸(CGA)在控制细胞凋亡的感知中起着广泛的作用。然而,CGA 在体外受 TD 影响的软骨细胞中的实际作用仍然未知。本研究通过 B 细胞淋巴瘤 2(Bcl-2)、Bcl-2 相关 X 蛋白(Bax)和 Caspase-3 与 CD147 信号研究 CGA 对软骨细胞凋亡的潜在作用。通过免疫荧光、western blot 分析和逆转录定量聚合酶链反应(RT-qPCR)研究这些标志物的表达。从胫骨生长板分离、培养和处理软骨细胞。使用亚致死剂量的 thiram(2.5μg/mL)诱导细胞毒性,然后用最佳剂量(40μg/mL)的 CGA 处理。结果表明,thiram 使软骨细胞变形,凋亡率增加。但是,在 CGA 的情况下,CD147 的高表达增强了软骨细胞的活力,同时下调了 Bax/Caspase-3 信号,上调了 Bcl-2。通过观察到细胞凋亡标志物 Caspase-3 在 CD147 过表达细胞中发生显著变化,排除了第一种可能性。相反,CD147 的免疫耗竭增强了 Caspase-3 的裂解,表明软骨细胞中凋亡的激活。因此,这些发现表明了 CD147 在 thiram 诱导的 TD 中关于 Bcl-2/Bax/Caspase-3 凋亡信号轴调节的新见解。

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