摘要
目的探讨苏木酮A(SA)对顺铂(DDP)引起人肾小管上皮(HK 2)细胞损伤的保护作用及其机制。方法将HK-2细胞随机分为5组:A组为仅加入相同体积二甲基亚砜(DMSO)溶液的对照组,B组为加入5 μmol/L的DDP组,C组为加入5 μmol/L DDP和10 μmol/L SA组,D组为加入 5 μmol/L DDP 和 20 μmol/L SA 组,E 组为加入 5 μmol/L DDP 和 30 μmol/L SA 组。采用嗟醴蓝(MTT)法检测HK-2细胞存活率,流式细胞术检测其凋亡率,酶联免疫吸附试验(ELISA)法检测细胞培养上清中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β蛋白水平,生物化学法检测细胞中丙二醛(MDA)、轻自由基(0H·)蛋白含量;蛋白质印记法(Western blot)检测磷酸化(p) p65、p-核因子抑制蛋白(P-IkB)蛋白表达水平及核转录因子E2相关因子2(Nrf2)核蛋白的表达水平。结果与B组比较,SA预处理各组(C、D、E组)HK-2细胞存活率均升高[(46. 23 ±6.52)%比(60.67 ±4.51)%.(66.56 ±6.58)%.(77.54 ±6.38)%,P<0.05],凋亡率均降低[(36.16 ±3.28)%比(29.43 ±1.31)%.(28.23 ±3.43)%.(21.04 ±3. 56)%,P <0.05],HK-2 细胞中炎症因子 TNF-a和IL-1β蛋白水平明显降低(P<0.05),MDA和OH·蛋白含量明显降低(P <0. 05),p-p65 ,p-IkB蛋白表达水平明显降低(P<0.05),Nrf2核蛋白表达水平明显升高(P<0.05).结论SA具有抗DDP所致HK-2细胞的细胞毒性作用,其机制可能与激活NH2通路或抑制通过NF-kB发挥抗氧化应激、抗炎及抗凋亡作用有关。
Objective To investigate the mechanism of sappanone A protecting human kidney tubular epithelial( HK-2) cells from cisplatin( DDP) induced injury. Methods HK-2 cells were randomly divided into 5 groups: Add the same volume of Dimethyl sulfoxide ( DMSO) as group A, add 5 μmol/L DDP as group B ,add 5 μmol/L DDP and 10μmol/L SA as group C ,add 5 μmol/L DDP and 20 μmol/L SA as group D,add 5 μmol/L DDP and 30 μmol/L SA as group E. MTT was used to detect the survival rate of HK-2 cells. Flow cytometry was used to detect the cell apoptosis rate. Enzyme-linked immunosorbent assay( ELISA) was used to detect the level of tumor necrosis factor( TNF)-α and interleukin ( IL)-1β protein in the supernatant of HK-2 cells. The protein level of Malondialdehyde ( MDA) and OH ? in HK-2 cells were detected by using colorimetry. Western blotting was used to detect the protein expression level of p-p65,p-IκB and nuclear factor related factor 2(Nif2). Result Compared with group B[(46.23 ±6.52)%], the survival rate of HK-2 cells increased in SA pretreatment groups[ group C,D,E were(60. 67 ±4. 51)%,(66.56 ±6.58)%,(77.54 ±6. 38)% respectively,P < 0. 05 ],the apoptosis rate of HK-2 cells decreased in SA pretreatment groups[ group C,D,E were(29.43 ±1.31)%,(2& 23 ± 3. 43)%,(21.04 ±3. 56)% respectively, P < 0. 05 ], the protein levels of TNF-a and IL-1 p decreased in SA pretreatment groups (P <0.05),the protein levels of MDA and OH ? decreased in SA pretreatment groups(P <0.05),the protein expression level of p p65 and p-IkB decreased in SA pretreatment groups ( P < 0. 05 ), the nucleprotein expression level of Nrf2 increased in SA pretreatment groups(P < 0.05). Conclusion SA has the cytotoxic effect against DDP on HK-2 cells. The mechanism may be related to the activation of Nrf2 pathway or the inhibition of antioxidant stress, anti-inflammatory and anti-apoptotic effects through NF-kB.
作者
康林
赵静
杨帆
徐明堂
王涛
赵焕芬
Kang Lin;Zhao Jing;Yang Fan;Xu Mingtang;Wang Tao;Zhao Huanfen(Department of Physiology,Hebei General Hospital, Shijiazhuang 050051, China)
出处
《临床内科杂志》
CAS
2019年第8期557-560,共4页
Journal of Clinical Internal Medicine
基金
河北省卫生和计划生育委员会医学科学研究重点课题计划(20170362).
