摘要
目的探讨红景天苷对小鼠心肌纤维化和焦亡的影响及潜在的作用机制。方法将小鼠随机分为对照组、模型组和红景天苷低、中、高剂量组,每组各10只。除对照组外,其余各组小鼠皮下注射异丙肾上腺素5 mg/(kg·d)造模。自造模之日起,红景天苷低、中、高剂量组小鼠每日灌胃红景天苷10、30、50 mg/kg,对照组和模型组小鼠每日灌胃生理盐水10 mL/kg,连续14 d。末次给药后处死小鼠,以苏木素-伊红染色法观察小鼠心肌组织病理变化并计算心肌细胞横径,Masson、Sirius Red染色法观察小鼠心肌组织纤维化程度并计算心肌胶原容积分数(CVF),实时荧光定量PCR法检测小鼠心肌组织中Ⅰ型胶原蛋白(ColⅠ)、α-平滑肌肌动蛋白(α-SMA)、Toll样受体4(TLR4)、NOD样受体蛋白3(NLRP3)、半胱氨酸蛋白酶1(caspase-1)、消皮素D(GSDMD)的mRNA表达水平,Western blot法和免疫组织化学法检测小鼠心肌组织中ColⅠ、α-SMA、TLR4、NLRP3、caspase-1、GSDMD的总蛋白表达水平及蛋白阳性细胞积分。结果与对照组比较,模型组小鼠心肌细胞体积增大,心肌纤维排列紊乱,细胞基质明显增加,心肌组织中CVF显著升高,ColⅠ、α-SMA、TLR4、NLRP3、caspase-1、GSDMD的mRNA、总蛋白表达水平及蛋白阳性细胞积分均显著升高(P<0.01)。与模型组比较,红景天苷中、高剂量组小鼠心肌细胞形态较清晰,心肌纤维化程度较低,心肌组织中以上指标水平均不同程度逆转,尤其是红景天苷高剂量组逆转程度最显著(P<0.05或P<0.01)。此外,红景天苷低剂量组部分纤维化和焦亡相关指标也在一定程度上逆转。结论红景天苷对心肌纤维化的发生、发展有明显的预防作用,其机制可能是通过抑制TLR4介导的心肌细胞焦亡通路激活。
OBJECTIVE To investigate the effects of salidroside(Sal)on myocardial fibrosis and pyroptosis and its potential mechanism.METHODS The mice were randomly divided into control group,model group and Sal low-dose,medium-dose and high-dose groups,with 10 mice in each group.Except for the control group,the mice in other groups were injected subcutaneously with isoproterenol 5 mg/(kg·d)to prepare the myocardial fibrosis model.Since modeling,mice in the Sal low-dose,medium-dose and high-dose groups were given 10,30 and 50 mg/kg of Sal by intragastric administration every day;control group and model group were given 10 mL/kg of normal saline by intragastric administration every day,for 14 consecutive days.After the last medication,the mice were sacrificed;hematoxylin-eosin staining was used to observe pathological change of myocardial tissue and calculate the diameter of myocardial cell;Masson and Sirius Red staining were used to observe the degree of myocardial fibrosis in mice and calculate the collagen volume fraction(CVF);quantitative real-time PCR was performed to detect the mRNA expressions of collagen typeⅠ(ColⅠ),α-smooth muscle actin(α-SMA),Toll-like receptor 4(TLR4),NOD-like receptor pyrin domain containing 3(NLRP3),caspase-1 and gasdermin D(GSDMD)in myocardial tissues.The total protein expressions of ColⅠ,α-SMA,TLR4,NLRP3,caspase-1 and GSDMD in myocardial tissues and protein-positive cell score were measured by Western blot assay and immunohistochemistry.RESULTS Compared with control group,the myocardial cells in the model group were enlarged,the arrangement of myocardial fibers was disordered,the matrix metabolism was significantly increased,the CVF in myocardial tissue was significantly increased,and the mRNA and protein expression levels of ColⅠ,α-SMA,TLR4,NLRP3,caspase-1 and GSDMD were elevated and protein-positive cell score was increased significantly(P<0.01).Compared with model group,the myocardial cell morphology was clearer,myocardial fibrosis was alleviated,and the levels of the above indi
作者
温方军
高磊
胡益敏
石开虎
WEN Fangjun;GAO Lei;HU Yimin;SHI Kaihu(Dept.of Cardio-Thoracic Surgery,Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Nanjing University of Chinese Medicine,Nanjing 210028,China;Jiangsu Province Academy of Chinese Medicine,Nanjing 210028,China;College of Traditional Chinese Medicine/College of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210046,China;Dept.of General Surgery,Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Nanjing University of Chinese Medicine,Nanjing 210028,China)
出处
《中国药房》
CAS
北大核心
2023年第9期1053-1059,共7页
China Pharmacy
基金
江苏省基础研究计划(自然科学基金)资助项目(No.BK20191503)
江苏省中医药研究院青年科学研究项目(No.QNKXYJ202109)。
关键词
红景天苷
心肌纤维化
焦亡
TOLL样受体4
NOD样受体蛋白3
salidroside
myocardial fibrosis
pyroptosis
Toll-like receptor 4
NOD-like receptor pyrin domain containing 3
