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P2X7R在小檗碱缓解小鼠慢性视网膜光损伤中的机制研究

Berberine Alleviates Chronic Retinal Light Damage in Mice: A Study of the Role of P2X7R and the Mechanisms Involved

  • 摘要:
    目的 探究P2X7R在小檗碱(berberine, BBR)缓解慢性视网膜光损伤小鼠中的动态变化趋势及相关作用机制。
    方法 90只小鼠随机分为空白对照组10只,LD组40只,LD+BBR组40只,后两组给予低能量(500 lux)蓝光照射(12 h/d)持续3个月,LD+BBR组在光照同时予200 mg/kg BBR灌胃。光照及灌胃结束后,取小鼠眼部组织,通过HE染色观察BBR对慢性视网膜光损伤小鼠的保护作用,TUNEL观察BBR对慢性视网膜光损伤小鼠凋亡细胞的影响,实时荧光定量PCR检测P2X7受体在BBR缓解慢性视网膜光损伤中的表达情况。
    结果 与空白对照组比较,LD组小鼠视网膜形态异常,部分神经节细胞核缩小,染色变深,排列松散;内核层细胞排列略松散;外核层细胞混乱排列,视网膜外核层厚度(47.11±2.01) μm;视网膜外核层出现大量凋亡细胞,凋亡率为(71.16±5.99)%,差异有统计学意义(P<0.05)。与LD组比较,LD+BBR组小鼠视网膜形态出现轻度异常,神经节细胞松散排序;视网膜内、外核层细胞形态略完整、染色均匀、排列紧密;外核层厚度为(54.07±2.05) μm,视网膜外核层有少量凋亡细胞,凋亡率为(16.02±2.68)%,差异有统计学意义(P<0.05)。与空白对照组比较,LD组小鼠视网膜中P2X7R mRNA的相对表达量上调,二者差异有统计学意义(P<0.05);LD+BBR组小鼠视网膜P2X7R mRNA的相对表达量下调,差异无统计学意义,但与LD组相比,视网膜P2X7R mRNA的相对表达量呈现明显下降趋势,且二者差异有统计学意义(P<0.05)。
    结论 BBR具有缓解小鼠慢性视网膜光损伤的作用,并抑制了P2X7R的激活进而阻止视网膜光损伤的形成。

     

    Abstract:
    Objective To investigate the trend of dynamic changes and the mechanisms of P2X7R by which berberine (BBR) alleviates chronic retinal light injury in mice.
    Methods A total of 90 mice were randomly divided into three groups, a blank control group (n = 10), a group exposed to low-intensity blue light (500 lux) for 12 hours per day for a duration of 3 months, which was referred to as the LD group (n = 40), and another group given BBR at a dose of 200 mg/kg via gastric gavage on top of the blue light exposure for the LD group, which was referred to as the LD + BBR group (n = 40). After the light exposure and gavage were completed, eye tissues were collected from the mice. Hematoxylin and eosin (HE) staining was performed to observe the protective effects of berberine on chronic retinal light damage in mice. TUNEL assay was performed to assess the effect of berberine on apoptotic cells in mice with chronic retinal light injury. Additionally, quantitative polymerase chain reaction (QPCR) was performed to assess the expression level of P2X7 receptors in chronic retinal light injury relieved by BBR.
    Results Compared with the blank control group, the LD group exhibited abnormal retinal morphology, with some ganglion cells displaying reduced nuclei, a deeper stain, and loose arrangement. In the LD group, the cells in the inner nuclear layer appeared to be slightly more loosely arranged, while the cells in the outer nuclear layer cells were arranged in a disorderly way. Furthermore, the thickness of the outer nuclear layer of the retina from mice in the LD group was (47.11 ± 2.01) μm, and a significant number of apoptotic cells were observed in the outer nuclear layer, resulting in an apoptosis rate of (71.16 ± 5.99)% (P < 0.05). In contrast, the LD + BBR group showed mild abnormal retinal morphology with loosely arranged ganglion cells. In the LD + BBR group, the cells in both inner and outer nuclear layers of retina exhibited relatively intact morphology, uniform staining pattern, and tight arrangement. Moreover, the thickness measurement for outer nuclear layer revealed a value of (54.07 ± 2.05) μm, and there were only a few apoptotic cells present, resulting in an apoptotic rate of (16.02 ± 2.68)% (P < 0.05). Compared with that of the blank control group, the relative expression of P2X7R mRNA in the retinas of the LD group was upregulated, with the difference between the two groups being statistically significant (P < 0.05). The relative expression of P2X7R mRNA in the retinas of the LD + BBR group was downregulated, showing no statistical significance compared with that of the blank control group. However, compared with that of the LD group, the relative expression of P2X7R mRNA in the retinas of the LD + BBR group showed a significant downward trend, and the difference between the two groups was statistically significant (P < 0.05).
    Conclusion Berberine can alleviate chronic retinal photodamage in mice, and inhibit the activation of P2X7R, thereby preventing the formation of retinal photodamage.

     

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