姜黄素通过Nrf2/HO-1通路缓解氟致BHK-21细胞氧化损伤的研究

Curcumin alleviates oxidative damage induced by sodium fluoride in BHK-21 cells via Nrf2/HO-1 signal pathway

本试验通过培养BHK-21细胞系,设定3个小组:对照组、NaF组、Cur5+NaF组。通过CCK-8法检测BHK-21细胞的增殖活性,借助微板法测定乳酸脱氢酶(LDH)的生成量。利用荧光探针DCFH-DA观察活性氧(ROS)的生成,用比色法检测丙二醛(MDA)和还原型谷胱甘肽(GSH)的含量以及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性,进一步采用荧光定量PCR和Western-blot评估Nrf2/HO-1通路相关基因与蛋白的表达水平。结果显示,NaF处理后,BHK-21细胞形态呈梭形,数量减少,LDH漏出率显著升高(P<0.01);氟处理显著增加了细胞中ROS和MDA的含量(P<0.01),降低了Nrf2、HO-1的表达水平以及SOD、GSH、GSH-Px的活性(P<0.01)。姜黄素处理后,BHK-21细胞排列紧密,边缘清晰,LDH漏出率显著下降(P<0.01);Nrf2和HO-1的表达水平显著增加,减少了ROS与MDA的生成(P<0.01),增强了SOD、GSH、GSH-Px的活性(P<0.01)。结果表明,姜黄素可以通过激活Nrf2/HO-1通路缓解NaF诱导的BHK-21细胞氧化损伤。

In the study,BHK-21 cell lines were cultured and set for three group:control group,NaF group and Cur5+NaF group.The proliferative activity of BHK-21 cells was detected by the CCK-8 method,and the production of LDH was measured with the microplate method.the production of ROS was observed using the fluorescent probe DCFH-DA,and the levels of MDA and GSH as well as the activities of SOD and GSH-Px was detected by the colorimetric method.The expression of Nrf2/HO-1 pathway-related genes and proteins were further assessed by real-time fluorescence PCR and Western blot,respectively.The results showed that after NaF treatment,the morphology of BHK-21 cells with spindle shaped,the number decreased,and the leakage rate of LDH significantly increase d(P<0.01).NaF treatment significantly increased the levels of ROS and MDA(P<0.01)and decreased the expression levels of Nrf2 and HO-1,as well as the activities of SOD,GSH and GSH-Px(P<0.01).After curcumin treatment,BHK-21 cells tightly arranged with clear edges and the LDH leakage rate was significantly decreased(P<0.01).the expression levels of Nrf2 and HO-1 significantly increased reducing the production of ROS and MDA(P<0.01),and enhancingthe activities of SOD,GSH,and GSH-Px(P<0.01).The results suggest that curcumin can alleviate NaF-induced oxidative damage of BHK-21 cells by activating the Nrf2/HO-1 pathway.