Effects of Changyuning Decoction on protein expression of claudin-1, ZO-1 and occludin in colonic tissue of rats with ulcerative colitis
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摘要:
目的 探讨肠愈宁对溃疡性结肠炎(UC)模型大鼠结肠组织紧密连接(TJ)关键蛋白咬合蛋白(occludin)、闭合蛋白-1(claudin-1)、带状闭合蛋白-1(ZO-1)表达的影响。 方法 将60只健康雄性SD大鼠按照随机数字表法分为空白对照组、模型对照组、肠愈宁组、美沙拉嗪组,每组各15只。采用2, 4, 6-三硝基苯磺酸(TNBS)/乙醇复合液(100 mg/kg TNBS+50%乙醇0.25 mL)一次性灌肠制备UC大鼠模型,造模成功后,药物干预7 d。光镜下观察大鼠结肠组织病理形态学变化,透射电镜观察大鼠结肠黏膜超微结构,免疫荧光染色法观察大鼠结肠组织occludin、claudin-1、ZO-1蛋白荧光强度。 结果 光镜与电镜观察显示,模型对照组大鼠结肠黏膜充血、水肿,炎症渗出明显,细胞间紧密连接断裂;而经肠愈宁干预后,大鼠结肠黏膜病理状态明显改善,细胞间紧密连接少部分断裂。免疫荧光染色结果显示,模型对照组大鼠结肠组织claudin-1、ZO-1、occludin蛋白荧光强度(0.11±0.01、0.11±0.01、0.09±0.01)较空白对照组明显降低(0.14±0.01、0.15±0.03、0.13±0.00,均P<0.05);经肠愈宁干预后,大鼠结肠组织claudin-1、ZO-1、occludin荧光强度较模型对照组明显升高(0.13±0.00、0.14±0.02、0.12±0.01,均P<0.05)。 结论 肠愈宁可通过上调TJ关键蛋白claudin-1、ZO-1、occludin的表达修复结肠黏膜损伤,治疗UC。 Abstract:Objective To investigate the effect of Changyuning Decoction on the expression of key proteins occludin, claudin-1 and zona occludens-1 (ZO-1) in the tight junction (TJ) of colonic tissue of model rats with ulcerative colitis (UC). Methods Sixty healthy male SD rats were randomly divided into the blank control group, model control group, Changyuning group and mesalazine group, with 15 rats in each group. The UC rat model was prepared by single enema with 2, 4, 6-trinitrobenzenesulfonic acid (TNBS)/ethanol complex solution (100 mg/kg TNBS+0.25 mL of 50% ethanol). After successful modelling, drug intervention was performed for 7 days. A light microscope was used to observe the pathomorphological changes of rat colon tissues, a transmission electron microscope was employed to observe the ultrastructure of rat colon mucosa, and immunofluorescence staining was utilised to determine the fluorescence intensities of occludin, claudin-1 and ZO-1 proteins in rat colon tissues. Results Light and electron microscopic observations showed that the colonic mucosa of UC model rats was congested and oedematous, with obvious inflammatory exudation and broken intercellular TJs. After Changyuning intervention, the pathological status of the colonic mucosa of rats was significantly improved, and a small part of intercellular TJs was broken. The results of immunofluorescence staining showed that the fluorescence intensities of claudin-1, ZO-1 and occludin protein in the colon tissue of UC model rats (0.11±0.01, 0.11±0.01, 0.09±0.01) were significantly lower than those in the blank control group (0.14±0.01, 0.15±0.03, 0.13±0.00, all P < 0.05). After Changyuning intervention, the fluorescence intensities of claudin-1, ZO-1 and occludin in the colon tissue of rats were significantly higher than those in the model control group (0.13±0.00, 0.14±0.02, 0.12±0.01, all P < 0.05). Conclusion Changyuning Decoction can repair colonic mucosal injury to treat UC by up-regulating the expression of key proteins of TJ including claudin-1, ZO-1 and occludin. -
Key words:
- Changyuning Decoction /
- Ulcerative colitis /
- Tight junction /
- Mechanical barrier
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表 1 各组大鼠claudin-1、ZO-1、occludin荧光强度比较(x ±s,IntDen/Area)
组别 只数 claudin-1 ZO-1 occludin 空白对照组 15 0.14±0.01 0.15±0.03 0.13±0.00 模型对照组 14 0.11±0.01a 0.11±0.01a 0.09±0.01c 美沙拉嗪组 14 0.12±0.01 0.12±0.01 0.10±0.00 肠愈宁组 14 0.13±0.00b 0.14±0.02b 0.12±0.01d F值 5.446 2.622 33.971 P值 0.025 0.123 <0.001 注:与空白对照组比较,aP<0.05,cP<0.01;与模型对照组比较,bP<0.05,dP<0.01。 -
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