DCFH DA staining showed that the proportion of ROS positive cells and the power of green inflorescence were somewhat improved in the existence of homocysteine 300 mM for 24 h. More over, Gemcitabine Gemzar therapy of BMSCs with homocysteine for 24 h surely could cause the obvious depolarization of mitochondrial membrane potential. These suggest that ROS mediated mitochondrial dysfunction is involved with homocysteine induced BMSCs apoptosis. We used two certain antioxidants DMTU and NAC, to confirm whether ROS is needed for homocysteine induced apoptosis of BMSCs. The increase of ROS in BMSCs was clearly improved by homocysteine 300 mM after treatment for 24 h, which can be effectively corrected by specific pre-treatment with DMTU and NAC, as shown in Figure 4a. AO/EB double staining also showed that NAC and DMTU Gene expression can reverse homocysteine induced apoptosis of BMSCs. More over, the depolarization of mitochondrial membrane potential induced by homocysteine was effortlessly reserved after pretreatment with NAC and DMTU for 24 h, suggesting ROS mediated mitochondrial membrane depolarization takes part in homocysteine induced the impairment of BMSCs. A big human body of evidence shows that MAPK signal pathway is involved in ROS mediated apoptosis. Nevertheless, whether MAPK indication process also plays a critical role in homocysteine caused BMSCs apoptosis remain unknown. Here, we discovered that the specific JNK chemical, SP600125 can reverse homocysteine induced BMSCs apoptosis featured from the inhibition of mitochondrial membrane potential depolarization and nucleus injury, minus the affect intracellular ROS level. Neither p38 MAKP inhibitor SB203580 or ERK inhibitor PD98059 has the capacity to change Avagacestat molecular weight homocysteine caused apoptotic morphological changes. These results indicate that JNK transmission pathway is needed for homocysteine caused BMSCs apoptosis. To ensure that JNK process contributed to homocysteineinduced BMSCs apoptosis, american blot was useful to find the appearance of JNK, p38 and ERK1/2, as well as p p53, caspase 3, cleaved caspase 3, Bcl 2 proteins in BMSCs with or without homocysteine 300 mM treatment. Amount 6a showed that homocysteine 300 mM can enhance phosphorylated JNK expression. Furthermore, homocysteine treatment didn’t notably change phosphorylated p38 and ERK1/2 protein expression in BMSCs. To be able to confirm that homocysteine induced BMSCs apoptosis, we also detected the expression of p p53, caspase 3, cleaved caspase 3 and Bcl 2 proteins after treatment. As shown in Figure 6b, homocysteine did not affect the expression of p p53, but increased cleaved caspase 3 expression. Bcl 2 was considerably reduced by therapy in BMSCs. We further investigate whether homocysteine therapy leads to the changes of BMSCs capabilities. The VEGF and IGF 1 levels in the culture medium of BMSCs before and after homocysteine therapy were determined by ELISA assay.