| 李珍,朱伟琴,俞瑾,等.通腑化瘀清胰汤对急性胰腺炎小鼠炎性因子和自噬功能的影响[J].浙江中医药大学学报,2024,48(12):1492-1499. |
| 通腑化瘀清胰汤对急性胰腺炎小鼠炎性因子和自噬功能的影响 |
| Effects of Tongfu Huayu Qingyi Decoction on Inflammatory Factors and Autophagy Function in Mice with Acute Pancreatitis |
| DOI：10.16466/j.issn1005-5509.2024.12.003 |
| 中文关键词: 急性胰腺炎 淀粉酶 小鼠 自噬 炎性因子 炎症反应 中草药 通腑化瘀清胰汤 |
| 英文关键词: acute pancreatitis amylase mice autophagy inflammatory factors inflammatory reaction Chinese herbal medicine Tongfu Huayu Qingyi Decoction |
| 基金项目:浙江省中医药科技计划科研基金项目(2022ZA137)；第七批全国老中医药专家学术经验继承工作项目(国中医药人教函〔2022〕76号) |
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| 中文摘要: |
| [目的] 研究通腑化瘀清胰汤对急性胰腺炎(acute pancreatitis，AP)小鼠炎性因子和自噬功能的影响。[方法] 40只C57BL/6小鼠随机分为空白对照组、AP模型组、通腑化瘀清胰汤组及奥曲肽组，每组10只。除空白对照组外，余下各组小鼠均使用雨蛙素(50 μg·kg-1)腹腔注射建立AP小鼠模型，空白对照组予等量0.9%氯化钠溶液腹腔注射。通腑化瘀清胰汤组小鼠予26 mL/(kg·d)通腑化瘀清胰汤灌胃，奥曲肽组小鼠背部皮下注射10 μg/(kg·d)奥曲肽注射液，治疗3 d；空白对照组、AP模型组予以观察处理。使用酶联免疫吸附分析(enzyme linked immunosorbent assay，ELISA)检测各组小鼠血清淀粉酶(amylase，AMY)、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素-6(interleukin-6，IL-6)及IL-10水平；苏木精-伊红(hematoxylin-eosin，HE) 染色法观察各组小鼠胰腺组织病理变化，电镜观察各组小鼠胰腺组织超微结构及自噬泡数量；免疫印迹法检测与自噬相关的蛋白自噬相关蛋白(Beclin1) 、选择性自噬接头蛋白(p62)及微管相关蛋白轻链3-Ⅱ(light chain 3-Ⅱ，LC3-Ⅱ)水平。[结果] 成功制备AP小鼠模型。与AP模型组比较，通腑化瘀清胰汤组及奥曲肽组小鼠血清中AMY、TNF-α、IL-6水平显著降低(P<0.05，P<0.01)，IL-10水平显著升高(P<0.01)。与AP模型组比较，通腑化瘀清胰汤组及奥曲肽组小鼠胰腺损伤情况和炎性细胞浸润情况均明显好转，HE评分均显著下降(P<0.05)，胰腺组织超微结构亦均有不同程度的恢复，自噬泡数量明显减少(P<0.05)。免疫印迹法显示，与空白对照组比较，AP模型组胰腺组织自噬相关标志物Beclin1、p62和LC3-Ⅱ蛋白表达水平显著升高(P<0.01)；与AP模型组比较，通腑化瘀清胰汤组及奥曲肽组胰腺组织Beclin1、p62和LC3-Ⅱ蛋白表达水平显著降低(P<0.05，P<0.01)。[结论] 通腑化瘀清胰汤能够降低AP小鼠血清AMY，调节炎性因子水平，减轻胰腺细胞自噬，从而缓解胰腺炎症反应和组织损伤。 |
| 英文摘要: |
| [Objective] To study the effects of Tongfu Huayu Qingyi Decoction on inflammatory factors and autophagy function in acute pancreatitis(AP) mice. [Methods] Total 40 C57BL/6 mice were randomly divided into blank control group， AP model group， Tongfu Huayu Qingyi Decoction group， and octreotide group， with 10 mice in each group. Except for blank control group， the remaining groups of mice were injected with caerulein(50 μg·kg-1) intraperitoneally to establish the AP mouse model. The blank control group was injected with an equal volume of 0.9% sodium chloride solution intraperitoneally. Mice in Tongfu Huayu Qingyi Decoction group were given 26 mL/(kg·d) Tongfu Huayu Qingyi Decoction by administration， mice in octreotide group were given 10 μg/(kg·d)octreotide injection subcutaneous injection on the back for 3 days， and blank control group and AP model group were observed. Serum amylase(AMY)， tumor necrosis factor-α (TNF-α)， interleukin-6(IL-6) and IL-10 levels were detected by enzyme-linked immunosorbent assay(ELISA). The pathological sections of pancreatic tissue of mice in each group were obtained by anatomy， and the pathological changes of pancreatic tissue were observed by hematoxylin-eosin(HE) staining， and the ultrastructure and number of autophagy vesicles were observed by electron microscopy. Western blot analysis was used to detect the levels of autophagy-related proteins， including Beclin-1， selective autophagy adaptor protein(p62)， and microtubule-associated protein light chain 3-Ⅱ(LC3-Ⅱ). [Results] AP mouse model was successfully prepared. Compared with AP model group， the levels of AMY， TNF-α and IL-6 in serum of Tongfu Huayu Qingyi Decoction group and octreotide group were significantly decreased (P<0.05， P<0.01)， and the content of IL-10 was significantly increased(P<0.01). Compared with AP model group， pancreatic injury and inflammatory cell infiltration in Tongfu Huayu Qingyi Decoction group and octreotide group were significantly improved， HE score was significantly decreased(P<0.05)， ultrastructure of pancreatic tissue was also recovered to varying degrees， and the number of autophagy vesicles was significantly reduced (P<0.05). Western blot showed that compared with blank control group， the protein expression levels of Beclin1， p62 and LC3-Ⅱ in pancreatic tissue of AP model group were significantly increased (P< 0.01). Compared with AP model group， Beclin1， p62 and LC3-Ⅱ protein expression levels in pancreas tissues of Tongfu Huayu Qingyi Decoction group and octreotide group were significantly decreased(P<0.05， P<0.01). [Conclusion] Tongfu Huayu Qingyi Decoction can reduce the serum AMY of AP mice， regulate the level of inflammatory factors， reduce the degree of autophagy of pancreatic cells ， thus reduce the pancreatic inflammation and tissue damage. |
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