ABSTRACT
Objective:To study the mechanism of Tiaogeng decoction in improving apoptosis of hypothalamic neurons through c-Jun N-terminal kinase (JNK) pathway. Method:The GT1-7 cell line of hypothalamic neuron cells treated by tributyltin chloride (TBTC) was used to induce a model of neuronal apoptosis, with 17β-estradiol as a positive drug. They were divided into control group, model group (1 mg·L-1), 17β-E2 group(100 nmol·L-1) and low, middle and high-dose Tiaogeng decoction groups (31.25,62.5,125 mg·L-1). The cell morphology was observed under a fluorescent inverted microscope, and cell counting kit-8 (CCK-8) was used to detect the cell viability. The cell nuclear morphology and the apoptosis rate of hypothalamic neurons were detected by Hoechst 33258 and Annexin V-FITC/propidium iodide(PI) double staining. Western blot was used to detect the expression levels of apoptosis signal-regulating kinase 1(ASK1), JNK, p53 and cleaved Caspase-3 of JNK pathway in GT1-7 cell. Western blot was used to analyze apoptosis-associated protein apoptosis signal-regulated kinase 1 (ASK1), JNK, p53, cleaved Caspase-3 expressions in JNK pathway and phosphorylation of JNK after intervention with JNK inhibitor SP600125, protein expression level of cleaved Caspase-3. Result:Compared with normal control group, the cell viability of the model group was significantly decreased (PPPPP-1)treatment significantly inhibited the expressions of p-JNK and cleaved Caspase-3 in GT1-7 cells (PConclusion:Tiaogeng decoction can improve the apoptosis of hypothalamic neurons through JNK pathway, and provide a theoretical basis for the treatment of menopausal syndrome and central nervous system protection.
ABSTRACT
Background and purpose:As a member of the catenin family, Delta-catenin protein could promote proliferation and invasion of tumor cells, but the accurate mechanism of Delta-catenin promoting cell proliferation is not clear. In the present study, we illustrated that Delta-catenin’s effect on cell apoptosis and their relationship with mitogen-activated protein kinase (MAPK) signaling pathway, and the possible mechanism was also explored for Delta-catenin promoting invasion and proliferation of tumor cells. Methods:The alterations of p38 and c-jun N-terminal rinasel JNK protein activity were detected in SPC and SK lung cancer cell lines with Delta-catenin overexpression or not, by Western blot method. At the same time, the apoptotic number of tumor cells was also examined by FCM method. Furthermore, the number of invasive tumor cells was examined by Matrigel invasive experiment. Results:Compared with untreated group and empty vector group, the activity of p38 protein was unchanged in lung cancer cell lines with Delta-catenin overexpressed (P>0.05), but the activity of JNK protein was decreased signiifcantly (P<0.05),meanwhile, apoptotic proportion of tumor cells were also reduced (P<0.05), and invasive ability of tumor cells was enhanced signiifcantly (P<0.05). Conclusion:Delta-catenin probably decreases apoptosis number of lung cancer cells via inhibiting the activity of JNK pathway, and then promotes invasive ability of tumor cells.