This study aims to review the protective effect and mechanism of carnosol on intestinal oxidative stress. by up-regulating the expression of Nrf2 and inhibiting p21 protein order AVN-944 to promote the expression of CCND1 and SOD. 0.05 was considered to be statistically significant. Results Carnosl protect ZYM-SIEC02 cells against t-BHP induced cell injury To measure the extent of t-BHP-induced damage to ZYM-SIEC02 cells, cell viability was detected. As shown in Physique 1A and ?and1B,1B, the order AVN-944 percentage of Edu positive cells in t-BHP treated group was lower than that of the control group ( 0.05). In carnosol treated group, ZYM-SIEC02 cells were pretreated with 10 M carnosol for 24 h and then treated with 200 M t-BHP for 3 h. The percentage of Edu positive cells in carnosol treated group was 88.95%, which was higher than that in the control group (82.64%) and t-BHP treated group (66.67%). Open in a separate window Physique 1 Carnosol protects ZYM-SIEC02 cells and reduces the effects of t-BHP on cell proliferation and viability. A. Edu staining results; B. Edu positive cell percentage statistics; C. MTT detection; D. CCK8 detection. We further verified the effects of t-BHP and carnosol on cells by MTT and CCK8 assays. The MTT test results showed that this OD values of the control group, t-BHP group, and carnosol group were 0.55, 0.08, and 0.56, respectively (Figure 1C). The CCK8 test results showed that this OD values of the control group, t-BHP group and carnosol group were 0.66, 0.05 and 0.72, respectively (Physique 1D). These results showed that t-BHP reduces cell proliferation and reduces cell order AVN-944 viability, while carnosol protects cells against t-BHP induced damage. Carnosol enhanced the ability of antioxidant in ZYM-SIEC02 cells Oxidative stress is an important mechanism of different type of cell damage. To clear the effect of carnosol on cellular oxidative stress, we examined the expression levels of ROS, MDA, SOD, and NO in three groups of cells. The results showed that this expression levels of ROS in the control group, t-BHP treatment group, and carnosol group were 24.32 RFU, 57.66 RFU, and 25.11 order AVN-944 RFU, respectively; the MDA expression levels were 0.2145 nM, 0.8744 nM, and 0.2454 nM; The expression levels of SOD were 50.57 U, 26.22 U, and 58.56 U, respectively; IL4R the expression levels of NO were 0.45 M, 0.95 M, and 0.47 M, respectively (Determine 2A-D). Our results showed that there was a significant increase in level of ROS, MDA, NO and decreased the production of SOD after treatment of t-BHP compared with the control group ( 0.05). We found that oxidative stress in ZYM-SIEC02 induced by t-BHP caused a cell damage and this condition alleviated by carnosol through regulating the content of several important antioxidant enzyme activities and key factors. Open in a separate window Number 2 Carnosic acid increases the antioxidant capacity of ZYM-SIEC02 cells. A. ROS; B. MDA; C. SOD; D. NO. Carnosol suppressed the oxidative stress by up-regulating the manifestation of Nrf2 and HO-1 We examined the manifestation of transcription factors related to oxidative stress, and recognized the expression levels of HO-1, FoxO3a, FoxM1, FoxO1, CDX2, E2F1, Nrf-2, and NF-B by q-PCR. The mRNA level of HO-1, Nrf2 were down-regulated after treatment of t-BHP compared with the control group ( 0.05). Moreover, pretreatment with carnosol could increase the expression level of HO-1, Nrf2 compared with the t-BHP group ( 0.05). These results showed that carnosol takes on an anti-oxidative part against t-BHP probably through up-regulating the manifestation of HO-1, Nrf2 to enhance antioxidant activities (Number 3A). Open in a separate windows Number 3 Detection of gene and protein manifestation by qPCR and western blot. A. qPCR results, * represents a significant difference between the t-BHP group and the control group; # represents a significant difference between the carnosol group and the.