白头翁汤正丁醇提取物通过激活BMP信号通路治疗溃疡性结肠炎的作用机制研究  

Mechanism of n-butanol extract of Pulsatilla Decoction in treating ulcerative colitis by activating BMP signaling pathway

作　　者：蒋晓娟 王亚东[1,2] 孙娟 马克龙[1,2] 吴大强[1,2] 邵菁 汪天明[1,2] 汪长中 JIANG Xiao-juan;WANG Ya-dong;SUN Juan;MA Ke-long;WU Da-qiang;SHAO Jing;WANG Tian-ming;WANG Chang-zhong(School of Integrated Traditional Chinese and Western Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;Institute of Integrated Traditional Chinese and Western Medicine,Anhui Academy of Chinese Medicine,Hefei 230012,China;School of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽省中医药科学院中西医结合研究所,安徽合肥230012 [3]安徽中医药大学中医学院,安徽合肥230012

出　　处：《中国中药杂志》2024年第7期1762-1773,共12页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82374173,81774034);安徽省重点研发计划项目(202104a07020020);安徽省高校自然科学研究重点项目(KJ2021A0590)。

摘　　要：该研究旨在探讨白头翁汤正丁醇提取物(n-butanol extract of Pulsatilla Decoction,BEPD)通过激活骨形态发生蛋白(BMP)信号通路发挥对溃疡性结肠炎(ulcerative colitis,UC)的治疗作用。C57BL/6小鼠分为6组:对照组、模型组、美沙拉嗪组及BEPD低、中、高剂量组。除对照组外,其余各组小鼠连续7 d自由饮用3%葡聚糖硫酸钠(dextran sulfate sodium,DSS)建立UC模型,然后进行不同浓度的BEPD和美沙拉嗪灌胃治疗。记录各组小鼠体质量和疾病活动指数(disease activity index,DAI)。处死小鼠后,取结肠组织进行组织学分析,采用阿利新蓝/过碘酸雪夫(AB/PAS)染色法检测杯状细胞的数量及黏液分泌状况,免疫组化法检测结肠组织中增殖标志物ki67、活化半胱天冬酶3(cleaved caspase-3)、粘蛋白2(mucin 2,Muc2)和基质金属蛋白酶9(MMP9)的表达,免疫荧光法检测结肠组织中紧密连接蛋白的表达,酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素(interleukin,IL)-1β和IL-6的水平,Western blot检测小鼠结肠组织中与BMP通路相关蛋白的表达,实时荧光定量PCR(qRT-PCR)法检测小鼠结肠杯状细胞分化相关基因表达。在体外细胞学实验中,进一步验证BEPD含药血清对脂多糖(LPS)诱导下的LS174T杯状细胞屏障功能的保护作用及其机制。结果表明,BEPD显著缓解UC小鼠的一般症状,恢复杯状细胞分化功能,促进Muc2分泌和紧密连接蛋白的表达,并抑制炎症因子的分泌,同时激活BMP信号通路。因此,该研究提示BEPD可能通过激活BMP信号通路发挥治疗UC的作用,为药物干预UC提供了新的思路。The study aimed to investigate the therapeutic effects of the n-butanol extract of Pulsatilla Decoction(BEPD)on ulcerative colitis(UC)via the bone morphogenetic protein(BMP)signaling pathway.C57BL/6 mice were divided into six groups:control,model,mesalazine,and BEPD low-,medium-,and high-dose groups.Except for the control group,the rest groups were treated with 3%dextran sulfate sodium(DSS)freely for seven consecutive days to establish the UC mouse model,followed by treatment with different concentrations of BEPD and mesalazine by gavage.The murine body weight and disease activity index(DAI)were recorded.After the mice were sacrificed,their colon tissues were collected for histological analysis.Alcian blue/periodic acid-Schiff(AB/PAS)staining was used to detect the number and mucus secretion status of goblet cells;immunohistochemistry was performed to measure the expression of ki67,cleaved caspase-3,mucin 2(Muc2),and matrix metalloproteinase-9(MMP9)in colon tissues;and immunofluorescence was used to analyze the expression of tight junction proteins in colon tissues,and enzyme linked immunosorbent assay(ELISA)was employed to quantify the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6.Western blot was conducted to evaluate the expression of BMP pathway-related proteins in mouse colon tissues.Quantitative real-time PCR(qRT-PCR)was performed to measure the expression of genes related to goblet cell differentiation in mouse colon tissues.In addition,this study also examined the protective effect and underlying mechanism of BEPD-containing serum on lipopolysaccharide(LPS)-induced barrier damages in LS174T goblet cells in vitro.The results showed that BEPD significantly alleviated UC symptoms in mice,restored goblet cell differentiation function,promoted Muc2 secretion and tight junction protein expression,and suppressed inflammatory factor secretion while activating the BMP signaling pathway.Therefore,BEPD may exert its therapeutic effects on UC by activating the BMP signaling pathway,providing

关 键 词：溃疡性结肠炎 白头翁汤正丁醇提取物 肠黏膜屏障 骨形态发生蛋白(BMP)信号通路 杯状细胞 

分 类 号：R285.5[医药卫生—中药学]