摘要
目的 探究芒柄花素(FMN)对炎症性肠病(IBD)大鼠肠上皮细胞凋亡的影响及可能机制。方法 采用三硝基苯磺酸诱导的方法构建大鼠IBD模型。将造模成功的48只大鼠按照随机数字表法分为模型组(生理盐水),低、高剂量FMN组(20、40 mg/kg FMN)和高剂量FMN+YAP抑制剂Verteporfin(VTPF)组(40 mg/kg FMN+10 mg/kg VTPF),每组12只;另取12只大鼠设为正常组(生理盐水);每天给药/生理盐水1次,连续7 d。末次给药后,对大鼠疾病活动指数(DAI)进行评分;测量大鼠结肠长度;观察大鼠结肠组织的病理学变化;检测大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10水平;检测大鼠肠上皮细胞的凋亡情况;检测大鼠结肠组织中Yes相关蛋白(YAP)、切割型胱天蛋白酶3(cleaved-caspase-3)、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达。结果 与正常组比较,模型组大鼠DAI评分,TNF-α、IL-6水平,肠上皮细胞凋亡率,cleaved-caspase-3、Bax蛋白表达水平均显著升高(P<0.05);结肠长度显著变短(P<0.05);IL-10水平和YAP、Bcl-2蛋白表达水平均显著降低(P<0.05);结肠组织出现细胞形态异常、排列紊乱,炎症细胞浸润等病理学变化。与IBD组比较,低剂量FMN组、高剂量FMN组大鼠上述指标水平均显著改善(P<0.05),且具有剂量依赖性(P<0.05);而加入VTPF后,显著逆转了FMN对IBD大鼠上述指标的改善作用(P<0.05)。结论 FMN可能是通过抑制Hippo/YAP信号通路,促进YAP的表达,进而抑制IBD大鼠肠上皮细胞凋亡。
OBJECTIVE To investigate the effects of formononetin(FMN)on the apoptosis of intestinal epithelial cells in inflammatory bowel disease(IBD)rats and its possible mechanism.METHODS IBD rat model was constructed by using trinitrobenzene sulfonic acid(TNBS)induction.Forty-eight rats with successful modeling were divided into model group(normal saline),low-dose and high-dose FMN groups(20 and 40 mg/kg FMN),and high-dose FMN+YAP inhibitor Verteporfin(VTPF)group(40 mg/kg FMN+10 mg/kg VTPF),with 12 rats in each group.Another 12 rats were set as the normal group(normal saline).They were given drug/normal saline,once a day,for 7 consecutive days.After the last administration,the disease activity index(DAI)of rats was calculated,and the colon length of rats in each group was measured.The pathological changes in the colon tissue of rats were observed.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-10 in serum were detected,and the apoptosis of intestinal epithelial cells was detected.The expressions of Yes associated protein(YAP),cleaved cysteine-containing aspartate proteolytic enzyme 3(cleaved-caspase-3),B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)were detected in colon tissue of rats.RESULTS Compared with the normal group,DAI score,the levels of TNF-αand IL-6,the apoptotic rate of intestinal epithelial cells,and the expressions of cleaved-caspase-3 and Bax protein in the model group were increased greatly(P<0.05);the length of the colon was greatly decreased(P<0.05),and the serum level of IL-10 and the protein expressions of YAP and Bcl-2 were greatly reduced(P<0.05).The cell morphology of colon tissue was abnormal,with disordered arrangement and inflammatory cell infiltration.Compared with IBD group,the above indexes of rats were improved significantly in low-dose and high-dose FMN groups(P<0.05),in dose-dependent manner(P<0.05).VTPF significantly alleviated the effects of FMN on the above indexes of IBD rats(P<0.05).CONCLUSIONS FMN may promote the expression of YAP by inhibiting the Hippo/YAP signaling pathway,thereby inhibiting apoptosis of intestinal epithelial cells in IBD rats.
作者
谢栋
刘媛媛
李正翔
袁恒杰
曹晓沧
XIE Dong;LIU Yuanyuan;LI Zhengxiang;YUAN Hengjie;CAO Xiaocang(Dept.of Pharmacy,Tianjin Medical University General Hospital,Tianjin 300052,China;Dept.of Genetics,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China;Dept.of Gastroenterology,Tianjin Medical University General Hospital,Tianjin 300052,China)
出处
《中国药房》
CAS
北大核心
2024年第13期1564-1569,共6页
China Pharmacy
基金
国家自然科学基金面上项目(No.82270565)。
