Article(id=1190375273666281481, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1190375270847710190, articleNumber=1001-2494(2025)03-0250-07, orderNo=null, doi=10.11669/cpj.2025.03.007, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1723478400000, receivedDateStr=2024-08-13, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1761737181345, onlineDateStr=2025-10-29, pubDate=1738944000000, pubDateStr=2025-02-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1761737181345, onlineIssueDateStr=2025-10-29, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1761737181345, creator=13701087609, updateTime=1761737181345, updator=13701087609, issue=Issue{id=1190375270847710190, tenantId=1146029695717560320, journalId=1190317699101192196, year='2025', volume='60', issue='3', pageStart='209', pageEnd='312', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1761737180673, creator=13701087609, updateTime=1761793989024, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1190613542412890252, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1190375270847710190, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1190613542412890253, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1190375270847710190, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=250, endPage=256, ext={EN=ArticleExt(id=1190375273922134030, articleId=1190375273666281481, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Effect and Mechanism of Saikosaponin A on Inflammatory Injury in Rats with Reflux Esophagitis, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

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.

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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。

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*胡以撒,男,学士,副主任医师 研究方向:消化内科
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杨芳,女,学士,主治医师 研究方向:反流性食管炎治疗

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杨芳,女,学士,主治医师 研究方向:反流性食管炎治疗

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J Agric Food Chem, 2022, 70(14):4353-4361., articleTitle=Chlorogenic acid alleviates chronic stress-induced duodenal ferroptosis via the inhibition of the IL-6/JAK2/STAT3 signaling pathway in rats, refAbstract=null)], funds=[Fund(id=1190958846374982527, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, awardId=2021RC124, language=CN, fundingSource=2021年浙江省卫生健康科技计划项目资助(2021RC124), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1190958842159706963, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, xref=1, ext=[AuthorCompanyExt(id=1190958842168095572, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, companyId=1190958842159706963, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 Zhejiang University of Traditional Chinese Medicine, Lishui 323400, China), AuthorCompanyExt(id=1190958842172289877, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, companyId=1190958842159706963, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 浙江中医药大学, 浙江 丽水 323400)]), AuthorCompany(id=1190958842235204438, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, xref=2, ext=[AuthorCompanyExt(id=1190958842243593047, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, companyId=1190958842235204438, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 General Medicine of the First Affiliated Hospital of Ningbo University, Ningbo 315201, China), AuthorCompanyExt(id=1190958842251981656, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, companyId=1190958842235204438, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 宁波大学附属第一医院全科医学, 浙江 宁波 315201)])], figs=[ArticleFig(id=1190958843589964654, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Fig.1, caption=Effects of SA on esophageal mucosal injury in RE rats. n=12, $\bar{x}\pm s$

A-results of esophageal mucosal damage in each group of rats; B-changes in the damage rate of esophageal mucosa in each group of rats;1)P<0.05, vs normal group;2)P<0.05, vs RE group; 3)P<0.05, vs SA high dose group.

, figureFileSmall=3O/mSbcVredgb/m3mlKzTw==, figureFileBig=Usdin1YoedLo3SPEQhv7qQ==, tableContent=null), ArticleFig(id=1190958843665462127, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=图1, caption=柴胡皂苷A(SA)对反流性食管炎(RE)大鼠食管黏膜损伤的影响。n=12, $\bar{x}\pm s$

A-各组大鼠食管黏膜损伤结果;B-各组大鼠食管黏膜损伤率变化;与正常组比较,1)P<0.05;与RE组比较,2)P<0.05;与SA高剂量组相比,3)P<0.05。

, figureFileSmall=3O/mSbcVredgb/m3mlKzTw==, figureFileBig=Usdin1YoedLo3SPEQhv7qQ==, tableContent=null), ArticleFig(id=1190958843745153904, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Fig.2, caption=Effects of SA on hematoxylin-eosin(HE) staining in esophageal tissue of RE rats, figureFileSmall=Z411513Ca3waf0bAkirv7A==, figureFileBig=kV2UBFeOYpknN6GDcnDB/w==, tableContent=null), ArticleFig(id=1190958843816457073, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=图2, caption=SA对RE大鼠食管组织苏木精-伊红(HE)染色结果的影响, figureFileSmall=Z411513Ca3waf0bAkirv7A==, figureFileBig=kV2UBFeOYpknN6GDcnDB/w==, tableContent=null), ArticleFig(id=1190958843883565938, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Fig.3, caption=Effect of SA on apoptosis of esophageal histopathocytes in RE rats. n=6, $\bar{x}\pm s$

A-TUNEL staining results of rat esophageal tissues in each group; B-changes in cell apoptosis rate in esophageal tissue of rats in each group; 1)P<0.05, vs normal group; 2)P<0.05, vs RE group; 3)P<0.05, vs 50 mg·kg-1 SA.

, figureFileSmall=JEiPOKybIop9jkn9EUCDpA==, figureFileBig=dFuwCeVjb6xArZEFgciBLA==, tableContent=null), ArticleFig(id=1190958845812945780, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=图3, caption=SA对RE大鼠食管组织细胞凋亡的影响。 n=6, $\bar{x}\pm s$

A-各组大鼠食管组织TUNEL染色结果;B-各组大鼠食管组织细胞凋亡率变化; 与正常组比较,1)P<0.05;与RE组比较,2)P<0.05;与50 mg·kg-1 SA相比,3)P<0.05。

, figureFileSmall=JEiPOKybIop9jkn9EUCDpA==, figureFileBig=dFuwCeVjb6xArZEFgciBLA==, tableContent=null), ArticleFig(id=1190958845901026166, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Fig.4, caption=Effects of SA on the immunohistochemical staining results of claudin-5 expression in esophageal tissue of RE rats. n=6, $\bar{x}\pm s$

A-immunohistochemical staining results of claudin-5 expression in the esophageal tissues of rats in each group; B-changes in average optical density values of claudin-5 in the esophageal tissues of rats in each group; 1)P<0.05, vs normal group; 2)P<0.05, vs RE group; 3)P<0.05, vs 50 mg·kg-1 SA.

, figureFileSmall=JQE9R2FJcDCw94/IsUfsbQ==, figureFileBig=FuXiIu0JTdob807CJJOuzg==, tableContent=null), ArticleFig(id=1190958845963940728, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=图4, caption=SA对RE大鼠食管组织中claudin-5表达的免疫组化染色结果的影响。n=6, $\bar{x}\pm s$

A-各组大鼠食管组织claudin-5表达免疫组化染色结果;B-各组大鼠食管组织中claudin-5平均光密度值的变化; 与正常组比较,1)P<0.05;与RE组比较,2)P<0.05;与50 mg·kg-1 SA相比,3)P<0.05。

, figureFileSmall=JQE9R2FJcDCw94/IsUfsbQ==, figureFileBig=FuXiIu0JTdob807CJJOuzg==, tableContent=null), ArticleFig(id=1190958846043632507, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Fig.5, caption=Effects of SA on the expression of IL-6, p-JAK2 and p-STAT3 proteins in esophageal tissue of RE rats. n=6, $\bar{x}\pm s$

1)P<0.05, vs normal group; 2)P<0.05, vs RE group; 3)P<0.05, vs 50 mg·kg-1 SA.

, figureFileSmall=T+i5mV1dmOtXgF5X5eNFJQ==, figureFileBig=UR7kPTFVW9gu2HzFE1DBmQ==, tableContent=null), ArticleFig(id=1190958846102352764, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=图5, caption=SA对RE大鼠食管组织中白细胞介素、磷酸化酪氨酸激酶2及信号转导和转录激活因子3蛋白表达的影响。 n=6, $\bar{x}\pm s$

与正常组比较,1)P<0.05;与RE组比较,2)P<0.05;与50 mg·kg-1 SA相比,3)P<0.05。

, figureFileSmall=T+i5mV1dmOtXgF5X5eNFJQ==, figureFileBig=UR7kPTFVW9gu2HzFE1DBmQ==, tableContent=null), ArticleFig(id=1190958846190433149, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=EN, label=Tab.1, caption=

Comparison of related inflammatory factors levels in rat esophageal tissue. n=6, $\bar{x}\pm s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Groups TNF-α/pg·mg-1 COX-2/pg·mg-1 IL-8/pg·mg-1
Normal 46.65±2.11 1.76±0.05 23.69±1.01
RE 90.86±3.431) 9.66±0.421) 51.52±2.081)
12.5 mg·kg-1 SA 72.25±3.022) 7.77±0.512) 43.35±1.762)
50 mg·kg-1 SA 53.36±2.182)3) 3.23±0.152)3) 29.93±1.252)3)
Omeprazole 51.99±2.062)3) 3.26±0.132)3) 30.06±1.372)3)
50 mg·kg-1 SA+rRIL-6 65.58±3.034) 6.57±0.184) 36.14±1.724)
), ArticleFig(id=1190958846253347710, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1190375273666281481, language=CN, label=表1, caption=

大鼠食管组织相关炎症因子水平比较。n=6, $\bar{x}\pm s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Groups TNF-α/pg·mg-1 COX-2/pg·mg-1 IL-8/pg·mg-1
Normal 46.65±2.11 1.76±0.05 23.69±1.01
RE 90.86±3.431) 9.66±0.421) 51.52±2.081)
12.5 mg·kg-1 SA 72.25±3.022) 7.77±0.512) 43.35±1.762)
50 mg·kg-1 SA 53.36±2.182)3) 3.23±0.152)3) 29.93±1.252)3)
Omeprazole 51.99±2.062)3) 3.26±0.132)3) 30.06±1.372)3)
50 mg·kg-1 SA+rRIL-6 65.58±3.034) 6.57±0.184) 36.14±1.724)
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柴胡皂苷A对反流性食管炎大鼠炎性损伤的影响及机制研究
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杨芳 1 , 胡以撒 2, *
中国药学杂志 | 论著 2025,60(3): 250-256
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中国药学杂志 | 论著 2025, 60(3): 250-256
柴胡皂苷A对反流性食管炎大鼠炎性损伤的影响及机制研究
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杨芳1, 胡以撒2, *
作者信息
  • 1 浙江中医药大学, 浙江 丽水 323400
  • 2 宁波大学附属第一医院全科医学, 浙江 宁波 315201
  • 杨芳,女,学士,主治医师 研究方向:反流性食管炎治疗

通讯作者:

*胡以撒,男,学士,副主任医师 研究方向:消化内科
Effect and Mechanism of Saikosaponin A on Inflammatory Injury in Rats with Reflux Esophagitis
Fang YANG1, Yisa HU2, *
Affiliations
  • 1 Zhejiang University of Traditional Chinese Medicine, Lishui 323400, China
  • 2 General Medicine of the First Affiliated Hospital of Ningbo University, Ningbo 315201, China
出版时间: 2025-02-08 doi: 10.11669/cpj.2025.03.007
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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。

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

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.

saikosaponin A  /  interleukin-6/tyrosine kinase 2 signal transducer  /  activator of transcription 3 pathway  /  reflux esophagitis  /  inflammation
杨芳, 胡以撒. 柴胡皂苷A对反流性食管炎大鼠炎性损伤的影响及机制研究. 中国药学杂志, 2025 , 60 (3) : 250 -256 . DOI: 10.11669/cpj.2025.03.007
Fang YANG, Yisa HU. Effect and Mechanism of Saikosaponin A on Inflammatory Injury in Rats with Reflux Esophagitis[J]. Chinese Pharmaceutical Journal, 2025 , 60 (3) : 250 -256 . DOI: 10.11669/cpj.2025.03.007
反流性食管炎(reflux esophagitis,RE)是指由食管下部损伤引起的食管炎症,导致胃内容物反流。伴随的胃内容物反流通常引起严重的炎症和溃疡,严重破坏正常的食管黏膜和组织,该疾病的特征是胸部烧灼感和餐后恶心[1]。目前,治疗RE最有效的药物主要是胃酸分泌抑制剂,包括H2受体拮抗剂(雷尼替丁)和质子泵抑制剂(奥美拉唑),虽然这些药物具有良好的治疗效果,但长期使用可能会引起肠道感染等副作用[2]。因此,寻找治疗RE疗效好、副作用小的天然药物是目前亟须解决的问题。柴胡皂苷A(saikosaponin A,SA)是从药用植物柴胡中提取的三萜皂苷,具有多种药理活性,包括抗炎、抗氧化等[3]。研究报道,SA可改善硫酸葡聚糖钠诱导的小鼠结肠炎[4]。但关于SA对RE大鼠炎性损伤的影响鲜有报道。相关研究显示,白细胞介素(interleukin,IL)-6/酪氨酸激酶2(janus kinase 2,JAK2)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)通路中的IL-6、磷酸化STAT3(p-STAT3)在RE大鼠食管组织中上调表达[5]。但SA是否可通过调控IL-6/JAK2/STAT3信号通路改善RE大鼠炎性损伤尚不可知。基于此,本研究拟通过构建RE大鼠模型,探究SA对RE大鼠炎性损伤的影响及其用机制。
72只,体质量为190~210 g的SPF级雄性SD大鼠[浙江中医药大学,生产许可证号:SCXK(浙)2021-0003]。本实验得到浙江中医药大学动物实验中心伦理委员会的批准,编号:2021-09101。
SA(批号20221203,上海源叶生物科技有限公司);IL-6激活剂重组大鼠IL-6蛋白(recombinant rat IL-6 protein,rRIL-6)(批号20221129,上海玉博生物科技有限公司);奥美拉唑(批号20221011,湖南方盛制药股份有限公司);大鼠肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、环氧化酶-2(cyclooxygenase-2,COX-2)、IL-8 ELISA试剂盒(批号20221003、20220929、20220917,北京方程嘉鸿科技有限公司);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)细胞凋亡试剂盒(批号20221118,上海康朗生物科技有限公司);兔源一抗紧密连接蛋白-5(claudin-5)、IL-6、p-JAK2、p-STAT3、GAPDH、JAK2、STAT3及羊抗兔二抗(批号ab131259、ab9324、ab219728、ab32143、ab8245、ab108596、ab68153、ab6721)(英国Abcam公司)。EOS R6 Mark Ⅱ型数码相机(日本佳能有限公司);RM2235切片机(德国徕卡公司);CX33型光学显微镜(日本奥林巴斯公司);4300酶标仪(美国Awareness公司);1658033型蛋白电泳仪(美国伯乐公司);Invitrogen iBright凝胶成像系统(美国赛默飞世尔科技有限公司)。
通过结扎前胃+部分结扎外置幽门法[6]构建RE模型,简言之,大鼠禁食不禁水24 h后腹腔注射质量分数3%戊巴比妥钠麻醉大鼠,开腹,游离胃部和幽门附近组织,用3-0线结扎前胃,用3-0线结扎幽门与0.38 mm玻璃棒后抽出玻璃棒,胃部放回原位,关腹。
SD大鼠随机分为RE组、正常组、SA低剂量组、SA高剂量组、奥美拉唑组、SA高剂量+rRIL-6组,每组12只。正常组大鼠经麻醉、开腹后将胃部提出腹腔并暴露5 min后,放回原位,关腹。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,建模操作完成后,对麻醉大鼠开腹,从胃食管交界上0.5 cm处截取2 cm食管组织,纵向切开食管,从宏观上观察RE大鼠食管黏膜表面存在黏膜病变、食管凹陷、出血等损伤,且微观上RE大鼠食管组织紧密性降低,且有大量炎性细胞浸润作为造模成功的标准。建模成功第二天开始给药,SA低剂量组、SA高剂量组[7]大鼠分别需灌胃12.5、50 mg·kg-1 SA,且均需腹腔注射等量的生理盐水;奥美拉唑组[8]大鼠需灌胃2 mg·kg-1奥美拉唑,且腹腔注射等量的生理盐水;SA高剂量+rRIL-6组[9]大鼠需灌胃50 mg·kg-1 SA且腹腔注射0.05 mg·kg-1rRIL-6;RE组、正常组大鼠需灌胃且腹腔注射等量的生理盐水。给药1天1次,持续2周。
末次处理结束后,麻醉大鼠,开腹,从胃食管交界上0.5 cm处截取2 cm食管组织,纵向切开食管以暴露内部损伤,并使用数码相机收集图像。使用Image J分析黏膜损伤率(公式1)。
食管黏膜损伤率%=食管损伤面积/食管总面积×100%
食管黏膜损伤率检测结束后,每组随机选取6只大鼠的食管组织,固定在体积分数4%多聚甲醛中,石蜡包埋,并切成4 μm厚的切片。切片进行HE染色。最后获取图像。
每组取剩余6只大鼠的食管组织,按ELISA试剂盒说明检测食管组织匀浆中TNF-α、COX-2、IL-8水平变化。
取“1.6”项中的食管组织切片常规脱蜡、水化、磷酸缓冲液(phosphate belanced solution,PBS)洗涤,用20 μg·mL-1蛋白酶K室温孵育30 min。PBS浸泡后,切片与TUNEL溶液在室温下孵育60 min。切片用苏木精染色后,用梯度乙醇脱水,二甲苯清洗2 min,中性胶封片。在光镜下对切片进行观察和拍照,利用Image J软件统计细胞凋亡率。
取“1.6”项中的食管组织切片,用质量分数0.3% Triton X-100和体积分数0.3%过氧化氢处理,以穿透细胞膜,阻断内源性过氧化物酶活性。再用牛血清白蛋白阻断30 min,加入claudin-5抗体(1∶600)在4 ℃下孵育过夜。第二天,用二抗孵育1 h,用新鲜二氨基联苯胺溶液染色。苏木精染色后,用梯度乙醇和二甲苯处理后,中性树脂封片。Image J软件计算claudin-5表达的平均光密度值。
放射免疫沉淀缓冲液用于从食管组织中分离蛋白质。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白质样品,然后转移到聚偏二氟乙烯膜上,质量分数5%脱脂牛奶封闭膜后,将膜与一抗IL-6(1∶4 000)、p-JAK2(1∶2 000)、p-STAT3(1∶5 000)、JAK2(1∶2 000)、GAPDH(1∶3 000)、STAT3(1∶5 000)在4 ℃下孵育过夜。第二天,将膜与二抗(1∶5 000)在室温下孵育1 h。洗涤后,添加化学发光试剂检测,并使用Image J定量分析蛋白灰度值。
GraphPad Prism9.0软件用于统计计算。计量资料表示为。单因素方差分析及事后SNK-q检验进行多组间比较,P<0.05为差异具有统计学意义。
正常组大鼠食管黏膜表面无损伤,RE组大鼠食管黏膜表面存在黏膜病变、食管凹陷、出血等损伤。与正常组相比,RE组大鼠食管黏膜损伤率升高(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黏膜损伤率降低(P<0.05);与SA高剂量组相比,SA高剂量+rRIL-6组大鼠食管黏膜损伤率升高(P<0.05),见图1
正常组大鼠食管组织结构正常,无明显炎症细胞浸润;RE组大鼠食管组织紧密性降低,且有大量炎性细胞浸润;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;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,见图2
与正常组相比,RE组大鼠食管组织中TNF-α、COX-2、IL-8水平升高(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中TNF-α、COX-2、IL-8水平降低(P<0.05);与SA高剂量组相比,SA高剂量+rRIL-6组大鼠食管组织中TNF-α、COX-2、IL-8水平升高(P<0.05),见表1
与正常组相比,RE组大鼠食管组织中细胞凋亡率升高(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中细胞凋亡率降低(P<0.05);与SA高剂量组相比,SA高剂量+rRIL-6组大鼠食管组织中细胞凋亡率升高(P<0.05),见图3
与正常组相比,RE组大鼠食管组织中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组大鼠食管组织中claudin-5平均光密度值降低(P<0.05),见图4
与正常组相比,RE组大鼠食管组织中IL-6、p-JAK2、p-STAT3蛋白表达升高(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中IL-6、p-JAK2、p-STAT3蛋白表达降低(P<0.05);与SA高剂量组相比,SA高剂量+rRIL-6组大鼠食管组织中IL-6、p-JAK2、p-STAT3蛋白表达升高(P<0.05),见图5
RE被认为是食道慢性炎症的结果,主要发生在食道组织暴露于胃酸反流时[10]。越来越多的研究表明,食管组织中炎性介质和炎性因子的表达参与了RE的进展,如RE大鼠食管组织中炎性介质COX-2和炎性因子TNF-α表达水平较高[11];促炎因子IL-8在RE大鼠体内处于高表达状态[12]。本研究通过结扎前胃+部分结扎外置幽门法构建了RE大鼠模型,从宏观上观察RE大鼠食管黏膜表面存在黏膜病变、食管凹陷、出血等损伤;从微观上观察RE大鼠食管组织紧密性降低,且有大量炎性细胞浸润,符合RE的特征。此外,本研究还发现RE组大鼠食管组织中TNF-α、COX-2、IL-8水平高于正常组,表明RE大鼠存在炎症反应。炎症与细胞凋亡密切相关,当过度炎症积累时,细胞凋亡呈上升趋势,且在慢性胃酸反流性食管炎大鼠食管中已观察到细胞大量凋亡的现象[13-14]。本研究也证明了RE大鼠食管中存在细胞异常凋亡。另有研究表明,当细胞凋亡增加时,食管黏膜完整性和屏障功能遭到破坏,claudin是一种完整的膜紧密连接蛋白,在上皮和内皮细胞的屏障功能中起重要作用,当食管黏膜发生病变时,claudin的表达异常,并显示出食管组织的严重损伤[15]。本研究中RE组大鼠食管组织中claudin-5平均光密度值低于正常组,表明RE大鼠存在食管黏膜屏障功能障碍。因此,降低炎症反应,抑制细胞凋亡,改善食管黏膜屏障功能障碍可能是治疗RE的有效途径之一。
SA是一种来源于柴胡的三萜皂苷,具有抗炎和抗氧化功能[16]。已有报道阐明,SA可抑制重症急性胰腺炎大鼠肠道炎症,并改善肠道屏障损伤[17]。本研究显示,低、高剂量SA均可抑制RE大鼠炎症反应及细胞凋亡,并改善食管黏膜屏障功能障碍。奥美拉唑是临床上常用于治疗RE的药物,本研究设置该药物为阳性药物,结果显示,高剂量SA与奥美拉唑对RE大鼠的治疗作用差异无统计学意义,提示SA可能成为治疗RE的潜在有效药物之一。长期服用奥美拉唑会出现腹胀、腹泻、恶心、呕吐、肝肾损伤等副作用,这就突出了中药SA在治疗RE中的优势。尽管SA毒副作用小,但大剂量使用也可能出现头晕、嗜睡、肝损伤、过敏反应等症状,因此,使用该药时要适时适量,谨遵医嘱。
IL-6是调节炎症反应的因素之一,其可通过调控下游因子JAK2/STAT3信号转导发挥促炎作用[18]。据报道,抑制IL-6/JAK2/STAT3通路可改善葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎[19];IL-6/JAK2/STAT3通路的阻断缓解了慢性应激诱导的大鼠十二指肠损伤[20]。本研究中,低、高剂量SA均抑制了RE大鼠食管组织中IL-6、p-JAK2、p-STAT3蛋白表达,由此可推测SA改善RE大鼠炎性损伤的机制可能与抑制IL-6/JAK2/STAT3通路有关。为了验证上述猜想,本实验用IL-6激活剂rRIL-6进行了回复实验,结果发现,rRIL-6逆转了高剂量SA对RE大鼠的影响,证实了SA改善RE大鼠炎性损伤的机制可能与部分抑制IL-6/JAK2/STAT3通路有关,而非阻断该通路。。
综上所述,SA可降低RE大鼠炎症反应,抑制细胞凋亡,改善食管黏膜屏障功能障碍,进而改善食管炎性损伤,该机制可能与抑制IL-6/JAK2/STAT3通路有关。未进一步探究具体是通过IL-6/JAK2/STAT3通路下游的哪些蛋白发挥作用是本研究设计的不足之一,后期将深入探讨。
  • 2021年浙江省卫生健康科技计划项目资助(2021RC124)
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2025年第60卷第3期
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doi: 10.11669/cpj.2025.03.007
  • 接收时间:2024-08-13
  • 首发时间:2025-10-29
  • 出版时间:2025-02-08
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  • 收稿日期:2024-08-13
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2021年浙江省卫生健康科技计划项目资助(2021RC124)
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    1 浙江中医药大学, 浙江 丽水 323400
    2 宁波大学附属第一医院全科医学, 浙江 宁波 315201

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*胡以撒,男,学士,副主任医师 研究方向:消化内科
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2种不同金属材料的力学参数

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属数
Number of
genus
种数
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species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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