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王月,刘丽芳,周亮,胡金辉.西黄胶囊调控IL-6/JAK2/STAT3信号通路促进非哺乳期乳腺炎大鼠模型创面愈合的作用机制研究[J].湖南中医药大学学报英文版,2023,43(7):1173-1179.[Click to copy
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| 西黄胶囊调控IL-6/JAK2/STAT3信号通路促进非哺乳期乳腺炎大鼠模型创面愈合的作用机制研究 |
| 王月,刘丽芳,周亮,胡金辉 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 观察西黄胶囊对非哺乳期乳腺炎大鼠模型创面愈合及白细胞介素-6（interleukin-6，IL-6）/非受体型酪氨酸蛋白激酶2（Janus kinase 2，JAK2）/信号转导及转录激活蛋白3（signal transducer and activator of transcription 3，STAT3）信号通路的影响。方法 复制非哺乳期乳腺炎大鼠模型，随机分为模型对照组、阳性药物组、中药治疗组、联合用药组，每组10只，另选取10只同批次SPF级健康SD大鼠作为假手术组。干预后观察并评估各组大鼠的创面愈合情况及乳腺炎症指数，HE染色法观察各组大鼠乳腺组织的病理改变；免疫组织化学法检测各组大鼠乳腺组织B淋巴细胞瘤-2基因关联X蛋白（B-cell lymphoma-2-associated X，BAX）、B淋巴细胞瘤-2基因（B-cell lymphoma-2，Bcl-2）的平均光密度值；Western blot法及RT-PCR法检测各组大鼠乳腺组织IL-6、JAK2、STAT3、人胱天蛋白酶9（Caspase-9）蛋白及mRNA的表达水平。结果 与假手术组大鼠比较，模型对照组大鼠的创面愈合率明显降低（P<0.05），而乳腺炎症指数，乳腺组织BAX、Bcl-2平均光密度值，IL-6、JAK2、STAT3、Caspase-9蛋白及mRNA的表达水平显著升高（P<0.05）。HE染色可见模型对照组大鼠乳腺腺叶存在明显的肉芽肿变性，周围充斥大量炎症细胞，阳性药物组、中药治疗组、联合用药组大鼠的病理情况存在不同程度的缓解，炎性细胞浸润及肉芽肿组织明显减少，“空泡样”坏死存在不同程度的愈合。与模型对照组比较，阳性药物组、中药治疗组、联合用药组的创面愈合率、Bcl-2光密度值均明显升高（P<0.05）；其中联合用药组的创面愈合率、Bcl-2光密度值明显高于阳性药物组及中药治疗组（P<0.05）；阳性药物组的创面愈合率高于中药治疗组（P<0.05），Bcl-2光密度值与中药治疗组比较差异无统计学意义（P>0.05）。与模型对照组比较，阳性药物组、中药治疗组、联合用药组的BAX光密度值、乳腺炎症指数均明显下降（P<0.05）；其中联合用药组的BAX光密度值、乳腺炎症指数明显低于阳性药物组及中药治疗组（P<0.05）；阳性药物组的乳腺炎症指数低于中药治疗组（P<0.05），BAX光密度值与中药治疗组比较差异无统计学意义（P>0.05）。与模型对照组比较，阳性药物组、中药治疗组、联合用药组的IL-6、JAK2、STAT3、Caspase-9蛋白及mRNA的表达水平均明显降低（P<0.05）；其中联合用药组上述蛋白及mRNA表达水平明显低于阳性药物组及中药治疗组（P<0.05），而阳性药物组的上述蛋白及mRNA表达水平均明显高于中药治疗组（P<0.05）。结论 西黄胶囊能有效促进非哺乳期乳腺炎大鼠模型的乳腺组织创面愈合，抑制乳腺组织细胞凋亡及炎症反应，其作用机制可能与西黄胶囊抑制IL-6/JAK2/STAT3信号通路相关。 |
| 关键词: 非哺乳期乳腺炎 西黄胶囊 IL-6/JAK2/STAT3信号通路 创面修复 作用机制 |
| DOI：10.3969/j.issn.1674-070X.2023.07.004 |
| Received:August 02, 2022 |
| 基金项目:湖南省中医药科研项目重点课题（2021009）；湖南省教育厅课题（19C1390）。 |
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| Mechanism of action for Xihuang Capsule in promoting wound healing in non-lactational mastitis rat model by regulating IL-6/JAK2/STAT3 signaling pathway |
| WANG Yue,LIU Lifang,ZHOU Liang,HU Jinhui |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To observe the effects of Xihuang Capsule on wound healing and the signaling pathway of interleukin-6 (IL-6)/Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) in non-lactational mastitis rat model. Methods The non-lactational mastitis rat model was duplicated and randomly divided into model control group, positive drug group, Chinese medicines group, and drug combination group, with 10 rats in each group. Another 10 SPF grade healthy SD rats in the same batch were selected as sham-operated group. After intervention, the wound healing condition and breast inflammation index of rats were observed and evaluated in each group, and histopathological changes of breast tissue of rats in each group were observed by HE staining. Immunohistochemical method was used to determine the average optical density of B-cell lymphoma-2 gene associated X protein (BAX) and B-cell lymphoma-2 gene (Bcl-2) in breast tissues of rats in each group. Western blot and RT-PCR were used to examine the expression levels of IL-6, JAK2, STAT3, human Caspase-9 protein, and mRNA in breast tissue of rats in each group. Results Compared with the rats in sham-operated group, the wound healing rate of rats in model control group was significantly lower (P<0.05), while the breast inflammation index, the average optical density of BAX and Bcl-2 in the breast tissue, and the expression levels of IL-6, JAK2, STAT3, Caspase-9 protein, and mRNA were significantly higher (P<0.05). HE staining showed that there was obvious granuloma degeneration in the mammary gland lobes of rats in model control group, surrounded by numerous inflammatory cells. The pathological condition of rats in positive drug group, Chinese medicines group, and drug combination group were alleviated to varying degrees. There was a significant decrease in inflammatory cell infiltration and granuloma tissue and the "vacuolar" necrosis was healed to varying degrees. Compared with model control group, the wound healing rate and Bcl-2 optical density of positive drug group, Chinese medicines treatment group, and drug combination group increased significantly (P<0.05). The wound healing rate and Bcl-2 optical density of drug combination group were significantly higher than those of positive drug group and Chinese medicines group (P<0.05). The wound healing rate of positive drug group was higher than that of Chinese medicines group (P<0.05), and there was no statistically significant difference in Bcl-2 optical density compared with Chinese medicines group (P>0.05). Compared with model control group, the BAX optical density and breast inflammation index of positive drug group, Chinese medicines group, and drug combination group decreased significantly (P<0.05). The BAX optical density and breast inflammation index of drug combination group were significantly lower than those of positive drug group and Chinese medicines group (P<0.05); the breast inflammation index of positive drug group was lower than that of Chinese medicines group (P<0.05) and the BAX optical density showed no statistical significance compared with that of Chinese medicines group (P>0.05). Compared with model control group, the expression levels of IL-6, JAK2, STAT3, Caspase-9 protein, and mRNA in positive drug group, Chinese medicines group, and drug combination group reduced significantly (P<0.05). The expression levels of the aforementioned proteins and mRNA in drug combination group were significantly lower than those in positive drug group and Chinese medicines group (P<0.05), while the expression levels of the aforementioned proteins and mRNA in positive drug group were significantly higher than those in Chinese medicines group (P<0.05). Conclusion Xihuang Capsule can effectively promote the wound healing of breast tissue in non-lactational mastitis rat model, and inhibit the apoptosis and inflammatory response of breast histiocyte. Its mechanism of action may be related to the inhibition of IL-6/JAK2/STAT3 signaling pathway by Xihuang Capsule. |
| Key words: non-lactational mastitis Xihuang Capsule IL-6/JAK2/STAT3 signaling pathway wound healing mechanism of action |
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