Icariin affects the expression of fractalkine and its receptor CX3CR1 in cerebral cortex of natural aging rat models

doi:10.3969/j.issn.2095-4344.0385

Abstract

Abstract:

BACKGROUND: Chemokine fractalkine not only involves chemotaxis, adhesion, and inflammation, but also participates in the proliferation and apoptosis of nerve cells in brain tissue.
OBJECTIVE: To observe the effect of icariin on the expression of fractalkine and its receptor CX3CR1 in cerebral cortex of natural aging rats.
METHODS: Eighty 3-month-old male Sprague-Dawley rats were randomly divided into four groups: young group, natural aging group, low- and high-dose icariin groups. Rats in the young and natural aging groups were intragastrically administered equal amount of normal saline for 60 days. Starting from 16 months old, rats in the drug groups were given 30 and 60 mg/kg icraiin for 60 days. Pathological changes in brain morphology were observed by hematoxylin-eosin staining. Brain senescence was identified by β-galactosidase staining. The expression of fractalkine and CX3CR1 was detected by immunohistochemistry. The levels of interleukin 6 and tumor necrotic factor α were tested by ELISA. The expression and activity of caspase-3 was determined by western blot assay and colorimetry, respectively.
RESULTS AND CONCLUSION: The expression level of fractalkine in the natural aging group was significantly lower than that in the young group. The expression of CX3CR1, the expression and activity of caspase-3, the number of positive particles of SA-β-gal, and the levels of interleukin 6 and tumor necrotic factor α in the natural aging group were significantly higher than those in the young group. After administration of icariin especially at high doses, the above phenomena were reversed. These results suggest that icariin can reduce the levels of inflammatory factors and inhibit apoptosis, which may be via regulating the expression of fractalkine and its receptor CX3CR1.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Epimedium, Aging, Chemotactic Factors, Caspase 3, Tissue Engineering

CLC Number:

R318

Zhang Xin1, Kang Zeng-jun2, Xue Juan1, Zhang Shun3, Liu Xiao-qing4, Liu Wei-hua1. Icariin affects the expression of fractalkine and its receptor CX3CR1 in cerebral cortex of natural aging rat models[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(32): 5203-5208.

Figures/Tables 2

2.1 实验动物数量分析实验选用雄性SD大鼠80只，分为4组，实验过程中无脱失，全部进入结果分析。 2.2 苏木精-伊红染色结果青年组胞浆染色浅，神经细胞大小均匀，胞核大而圆，核形规则。自然衰老组与青年组相比胞浆深染，神经元体积缩小，核固缩，核形不规则。与自然衰老组相比，淫羊藿苷低剂量组胞浆染色略均匀，神经元体积有所增大，核形略规则；淫羊藿苷高剂量组-伊红染色是最常规的病理切片染色，从大脑皮质神经细胞形态上看，淫羊藿苷对脑组织细胞图1。结构有一定的改善作用，见胞浆染色较均匀而色浅，神经元体积增大，可见少量固缩细胞，核形较规则。苏木精 2.3 SA-β-gal染色结果 SA-β-gal染色阳性产物为蓝色颗粒，各实验组大鼠大脑皮质均有阳性颗粒。与青年组相比，自然衰老组大鼠大脑皮质阳性颗粒显著增多，差异有显著性意义(P < 0.01)。与自然衰老组相比，淫羊藿苷低、高剂量组大鼠大脑皮质阳性颗粒减少，差异有显著性意义(P < 0.01)。与低剂量组相比，高剂量组大鼠大脑皮质阳性颗粒减少明显，差异有显著性意义(P < 0.05)。SA-β-gal染色法是鉴定细胞衰老的重要生物学标志[15]。自然衰老大鼠大脑皮质阳性颗粒数显著增多；淫羊藿苷可以减少大鼠大脑皮质阳性颗粒数。见图2。 2.4 趋化因子Fractalkine、CX3CR1免疫组织化学结果趋化因子Fractalkine免疫组织化学阳性反应为棕色或棕黄色颗粒定位于细胞膜，各实验组大鼠大脑皮质均有阳性表达。与青年组相比，自然衰老组大鼠大脑皮质阳性表达减少，差异有显著性意义(P < 0.01)。与自然衰老组相比，淫羊藿苷低、高剂量组大鼠大脑皮质阳性表达增多，差异有显著性意义(P < 0.01)，见图3。 CX3CR1免疫组织化学阳性反应为棕色或棕黄色颗粒定位于细胞膜，各实验组大鼠大脑皮质均有阳性表达。与青年组相比，自然衰老组大鼠大脑皮质阳性表达增多，差异有显著性意义(P < 0.01)。与自然衰老组相比，淫羊藿苷低、高剂量组大鼠大脑皮质阳性表达减少，差异有显著性意义(P < 0.01)，见图4。 2.5 脑组织白细胞介素6、肿瘤坏死因子α水平检测结果与青年组相比，自然衰老组大鼠大脑皮质中白细胞介素6、肿瘤坏死因子α水平显著升高，差异有显著性意义(P < 0.01)；与自然衰老组相比，淫羊藿苷低、高剂量组大鼠大脑皮质中白细胞介素6、肿瘤坏死因子α水平明显降低，差异有显著性意义(P < 0.01)，见图5。 2.6 WesternBlot法测定Caspase-3蛋白表达Western-Blot检测各组大鼠大脑皮质Caspase-3 蛋白水平，β-actin作为内参照。与青年组相比，自然衰老组大鼠大脑皮质中Caspase-3蛋白水平显著升高，差异有显著性意义(P < 0.01)；与自然衰老组相比，淫羊藿苷低剂量组大鼠大脑皮质中Caspase-3的蛋白水平虽有降低，但是差异无显著性意义(P > 0.05)；淫羊藿苷高剂量组大鼠皮质中Caspase-3的蛋白水平明显降低，差异有显著性意义(P < 0.05)，见图6。"

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