文献类型：期刊文献

中文题名：当归红芪超滤物对H9C2心肌细胞氧糖剥夺/恢复诱导细胞凋亡的干预作用

英文题名：Intervention of ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix on apoptosis induced by oxygen and glucose deprivation/recovery of H9C2 cardiomyocytes

作者：王艳平[1];赵信科[1];蒋虎刚[1];王新强[1];孔珊[1];韩金晏[1];李应东[2]

第一作者：王艳平

机构：[1]甘肃中医药大学中西医结合学院,甘肃兰州730000;[2]甘肃中医药大学附属医院心血管一区,甘肃兰州730000

第一机构：甘肃中医药大学中西医结合学院

年份：2022

卷号：38

期号：9

起止页码：899

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金会地区基金资助项目(81860786);甘肃省中医药研究中心基金资助项目(zyzx-2020-zy9,zyzx-2020-zx5);甘肃省中医药防治慢性疾病重点实验室开放基金资助项目(zyzx-2020-35)。

语种：中文

中文关键词：当归红芪超滤物;H9C2心肌细胞;氧糖剥夺/恢复;细胞凋亡;尼可地尔;肌钙蛋白T;C-反应蛋白

外文关键词：ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix;H9C2 cardiomyocytes;oxygen and glucose deprivation/recovery;cell apoptosis;nicorandil;troponin T;C-reactive protein

摘要：目的 探讨当归红芪超滤物对H9C2心肌细胞氧糖剥夺/恢复损伤的影响,并探索其可能机制。方法 H9C2心肌细胞进行氧糖剥夺处理6 h,建立心肌细胞缺血再灌注损伤模型。成年Wistar大鼠口服灌胃给药制备药物血清。H9C2心肌细胞造模处理后分为4组:模型组、阳性对照组、当归红芪超滤物组和联合组。空白组为正常H9C2细胞,模型组、阳性对照组、当归红芪超滤物组、联合组分别使用完全培养基、7.5 mg·kg^(-1)·d^(-1)尼可地尔药物血清培养基、10 mg·kg^(-1)·d^(-1)当归红芪超滤物药物血清培养基和联合药物血清(7.5 mg·kg^(-1)·d^(-1)尼可地尔+10 mg·kg^(-1)·d^(-1)当归红芪超滤物)培养基在常氧条件下培养4 h。用酶联免疫吸附实验测定肌钙蛋白T、肌钙蛋白Ⅰ及C-反应蛋白含量;用实时定量-聚合酶链反应检测B淋巴细胞瘤-2(Bcl-2)和胱天蛋白酶-3(Caspase-3)的基因表达;用5-乙炔基-2′-脱氧尿苷(EdU)荧光染色法检测细胞增殖;用激光共聚焦显微镜检测Bcl-2、 Caspase-3相关蛋白的表达。结果 空白组、模型组、当归红芪超滤物组、阳性对照组、联合组的肌钙蛋白T含量分别为(11.72±1.13),(99.34±14.85),(68.50±10.42),(65.50±9.98)和(54.83±9.08)ng·L^(-1);肌钙蛋白Ⅰ含量分别为(7.53±0.59),(38.33±3.94),(26.17±6.67),(23.17±4.84)和(19.83±2.97)ng·L^(-1);C-反应蛋白含量分别为(3.12±0.80),(32.17±4.56),(20.67±2.49),(18.17±3.48)和(13.83±2.67)mg·L^(-1);EdU阳性率分别为(69.61±5.03)%,(31.66±1.90)%,(55.81±2.11)%,(57.47±3.67)%和(67.86±4.54)%;Caspase-3基因表达水平分别为0.80±0.13,2.81±0.56,2.03±0.12,2.01±0.12和1.49±0.35;Bcl-2基因表达水平分别为1.56±0.09,1.07±0.13,1.24±0.07,1.24±0.13和1.28±0.12;Bcl-2和Caspase-3相关蛋白的表达的趋势与基因一致。上述指标,模型组与空白组相比,或当归红芪超滤物组、阳性对照组、联合组与模型相比,差异均有统计学意义(均P<0.05)。结论 当归红芪超滤物降低了H9C2心肌细胞氧糖剥夺/恢复刺激后的细胞损伤、细胞凋亡,缓解了心肌细胞氧糖剥夺/恢复损伤。
Objective To study ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix (RAS-RH) content on H9C2 cardiomyocytes oxygen and glucose deprivation / recovery damage,and explore its possible mechanism. Methods H9C2 cardiomyocytes were treated with oxygen and glucose deprivation for 6 h to establish myocardial ischemia reperfusion injury model.Adult Wistar rats were orally administered intragastrically to prepare drug serum. H9C2 cardiomyocytes were divided into 4 groups after modeling: model group,positive control group,RAS-RH group and combine group;the blank group was normal H9C2 cells. The model group,positive control group,RAS-RH group and combine group were cultured in complete medium,7. 5 mg ·kg^(-1)·d^(-1)nicorandil drug serum medium,10 mg ·kg^(-1)·d^(-1)RAS-RH ultrafiltration drug serum medium and combined drug serum (7. 5 mg·kg^(-1)·d^(-1)nicorandil + 10 mg·kg^(-1)·d^(-1)RAS-RH ultrafiltration) medium under normoxia for 4 h,respectively. Enzyme-linked immunosorbent assay was used to determine the contents of troponin T,troponin I and C-reactive protein;real-time quantitative polymerase chain reaction was used to detect the gene expression of B cell lymphoma 2 (Bcl-2) and Caspase-3;EdU fluorescence staining was used to detect cell proliferation;the expression of Bcl-2 and Caspase-3 related proteins was detected by laser confocal microscopy. Results The blank group,model group,RAS-RH group,positive control group and combine group of troponin T levels were (11. 72±1. 13) ,(99. 34±14. 85) ,(68. 50±10. 42) ,(65. 50±9. 98) ,and (54. 83±9. 08) ng·L^(-1);troponin Ⅰ protein content were(7. 53±0. 59) ,(38. 33±3. 94) ,(26. 17±6. 67) ,(23. 17±4. 84) ,and (19. 83±2. 97) ng·L^(-1);C-reactive protein content were (3. 12±0. 80) ,(32. 17±4. 56) ,(20. 67±2. 49) ,(18. 17±3. 48) ,and (13. 83±2. 67);positive rate of EdU cells were(69. 61±5. 03) % ,(31. 66±1. 90) % ,(55. 81±2. 11) % ,(57. 47±3. 67) % ,and (67. 86±4. 54) %;Caspase-3gene expression were 0. 80±0. 13,2. 81±0. 56,2. 03±0. 12,2. 01±0. 12,and 1. 49±0. 35;Bcl-2 gene expression were 1. 56±0. 09,1. 07±0. 13,1. 24±0. 07,1. 24±0. 13,and 1. 28±0. 12. The trend of Bcl-2 and Caspase-3 related protein expression was consistent with its gene expression. The above indicators: model group compared with blank group,positive control group,RAS-RH group and combine group compared with the model group,the difference had statistical significance (P < 0. 05) . Conclusion RAS-RH decreased H9C2Cardiomyocytes oxygen and glucose deprivation / recovery after stimulation of cell damage,cell apoptosis,and relieve the myocardial damage of oxygen and glucose deprivation / recovery.