目的 探讨柴胡皂苷A(saikosaponin A,SA)调节白细胞介素(interleukin,IL)-6/酪氨酸激酶2(tyrosine kinase 2,JAK2)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)通路对反流性食管炎(reflux esophagitis,RE)大鼠炎性损伤的影响。方法 SD大鼠随机分为RE组、正常组、SA低剂量组(灌胃12.5 mg·kg-1 SA)、SA高剂量组(灌胃50 mg·kg-1 SA)、奥美拉唑组(灌胃2 mg·kg-1奥美拉唑)、SA高剂量+IL-6激活剂重组大鼠IL-6蛋白(recombinant rat IL-6 protein,rRIL-6)组(灌胃50 mg·kg-1 SA+腹腔注射0.05 mg·kg-1 rRIL-6),每组12只。除正常组外,其他组大鼠均需通过结扎前胃+部分结扎外置幽门法构建RE模型,建模成功第二天开始给药,给药1天1次,持续2周。检测大鼠食管黏膜损伤率;苏木精-伊红染色(hematoxylin-eosin staining,HE)检测食管组织的病理学变化;酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测食管组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、环氧化酶-2(cyclooxygenase-2,COX-2)、IL-8水平;末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(terminal dexynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)检测食管组织中的细胞凋亡;免疫组化染色检测食管组织中紧密连接蛋白-5(claudin-5)表达;Western blot检测食管组织中IL-6、磷酸化JAK2(p-JAK2)、p-STAT3蛋白。结果 与正常组相比,RE组大鼠食管组织紧密性降低,且有大量炎性细胞浸润,食管黏膜损伤率、食管组织中TNF-α、COX-2、IL-8水平、细胞凋亡率、IL-6、磷酸化-JAK2(p-JAK2)、p-STAT3蛋白表达升高,食管组织中claudin-5平均光密度值降低(P<0.05);与RE组相比,SA低剂量组、SA高剂量组、奥美拉唑组大鼠食管组织病理损伤减轻,食管黏膜损伤率、食管组织中TNF-α、COX-2、IL-8水平、细胞凋亡率、IL-6、p-JAK2、p-STAT3蛋白表达降低,食管组织中claudin-5平均光密度值升高(P<0.05);与SA高剂量组相比,SA高剂量+rRIL-6组大鼠食管组织病理损伤严重,食管黏膜损伤率、食管组织中TNF-α、COX-2、IL-8水平、细胞凋亡率、IL-6、p-JAK2、p-STAT3蛋白表达升高,食管组织中claudin-5平均光密度值降低(P<0.05)。结论 SA可能通过抑制IL-6/JAK2/STAT3信号通路改善RE大鼠炎性损伤。
Abstract
OBJECTIVE To investigate the effect of saikosaponin A (SA) on inflammatory injury in rats with reflux esophagitis (RE) by regulating the interleukin (IL)-6/tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway. METHODS SD rats were randomly divided into RE group, normal group, SA low-dose group (gavage of 12.5 mg·kg-1 SA), SA high-dose group (gavage of 50 mg·kg-1 SA), omeprazole group (gavage of 2 mg·kg-1 omeprazole), SA high-dose+IL-6 activator recombinant rat IL-6 protein (rRIL-6) group (gavage of 50 mg·kg-1 SA+intraperitoneal injection of 0.05 mg·kg-1 rRIL-6), with 12 rats in each group. Except for the normal group, rats in all other groups were required to undergo RE model construction through fore-stomach ligation combined with partial ligation of the external pylorus. After successful modeling, the drug was administered once a day for 2 weeks. The damage rate of esophageal mucosa was detected. Hematoxylin-eosin staining(HE) was applied to detect pathological changes in esophageal tissue. Enzyme-linked immunosorbent assay (ELISA) was applied to detect levels of tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), and IL-8 in esophageal tissue. Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was applied to detect cell apoptosis in esophageal tissue. Immunohistochemical staining was applied to detect the expression of claudin-5 in esophageal tissue. Western blot was applied to detect IL-6, p-JAK2, and p-STAT3 proteins in esophageal tissue. RESULTS Compared with the normal group, the esophageal tissue compactness of rats in the RE group decreased, and there was a large amount of inflammatory cell infiltration, the incidence of esophageal mucosal injury, levels of TNF-α, COX-2, IL-8 in esophageal tissue, apoptosis rate, and the expression of IL-6, p-JAK2, and p-STAT3 proteins increased, while the average optical density of claudin-5 in esophageal tissue decreased (P<0.05). Compared with the RE group, the pathological damage to the esophageal tissue of rats in the SA low-dose group, SA high-dose group, and omeprazole group was reduced, the incidence of esophageal mucosal injury, levels of TNF-α, COX-2, IL-8 in esophageal tissue, apoptosis rate, and the expression of IL-6, p-JAK2, and p-STAT3 proteins decreased, while the average optical density of claudin-5 in esophageal tissue increased (P<0.05). Compared with the SA high-dose group, the SA high-dose+rRIL-6 group had severe pathological damage to the esophageal tissue, the incidence of esophageal mucosal injury, levels of TNF-α, COX-2, IL-8 in esophageal tissue, apoptosis rate, and the expression of IL-6, p-JAK2, and p-STAT3 proteins increased, while the average optical density of claudin-5 in esophageal tissue decreased (P<0.05). CONCLUSION SA may improve inflammatory injury in RE rats by inhibiting the IL-6/JAK2/STAT3 signaling pathway.
关键词
柴胡皂苷A /
白细胞介素-6/酪氨酸激酶2信号转导 /
转录激活因子3通路 /
反流性食管炎 /
炎症
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Key words
saikosaponin A /
interleukin-6/tyrosine kinase 2 signal transducer /
activator of transcription 3 pathway /
reflux esophagitis /
inflammation
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中图分类号：
R966
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脚注
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基金
2021年浙江省卫生健康科技计划项目资助(2021RC124)
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