1.Effect and Underlying Mechanism of Harmine on Proliferation and Apoptosis of Gastric Cancer Cells
Ting ZHANG ; Shiye JIANG ; Xingxing JIN ; Wenling ZHANG ; Na YU ; Xiaolin LI ; Guoqin ZHU ; Yichan ZHOU ; Yun SHAO ; Weihao SUN
Chinese Journal of Gastroenterology 2018;23(4):221-225
Background:Previous study has found that harmine inhibited the proliferation of gastric cancer cells by down-regulating cyclooxygenase-2(COX-2)expression. However,its molecular mechanism is not fully clear. Aims:To investigate the effect of harmine on proliferation and apoptosis of gastric cancer cells,and explore the role of PTEN/Akt/MDM2 signaling pathway in this process. Methods:Human gastric adenocarcinoma cell line SGC-7901 and MKN-45 were treated with harmine at different concentrations(2,4,8,16,32 μg/mL)for 24,48,and 72 hours. The cell proliferation and apoptosis were detected by MTT assay and Hoechst staining,respectively. The expressions of PTEN,COX-2, phosphorylated Akt(p-Akt)and p-MDM2 were measured by Western blotting. Results:Harmine dose- and time-dependently inhibited proliferation and induced apoptosis of SGC-7901 and MKN-45 cells. Also,harmine dose-dependently increased PTEN expression,and inhibited p-Akt,p-MDM2 and COX-2 expressions in SGC-7901 and MKN-45 cells. Conclusions:Harmine may inhibit proliferation and induce apoptosis of gastric cancer cells via down-regulating COX-2 expression through PTEN/Akt/MDM2 signaling pathway.
2.Effects of Tuina on P2Y12/RhoA/ROCK2 Pathway and c-Fos Protein Expression of Microglia in Spinal Dorsal Horn of Sciatic Nerve Chronic Compressive Injury Rats
Jingjing JIANG ; Zhigang LIN ; Hongye HUANG ; Huanzhen ZHANG ; Lechun CHEN ; Hui LIN ; Shiye WU ; Shuijin CHEN
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(7):100-105
Objective To observe the effects of tuina on P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression of microglia in spinal dorsal horn of sciatic nerve chronic compressive injury(CCI)rats;To explore the mechanism of tuina in treating lumbar disc herniation.Methods A CCI model of right sciatic nerve was used to simulate neuropathic pain in lumbar disc herniation.24 male SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.After 4 days of modeling,massage intervention was used to in the tuina group for 14 days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats before and on the 4th,10th,and 17th day after modeling were observed,immunofluorescence staining was used to detect the expression of Iba1 and P2Y12 protein in right spinal dorsal horn,Western blot was used to detect the expressions of RhoA and ROCK2 protein in right spinal dorsal horn,immunohistochemistry staining was used to detect the number of c-Fos positive cells in right spinal dorsal horn.Results Compared with the blank group,the PWT and PWL of the model group were significantly reduced on the 4th,10th,and 17th day after modeling(P<0.001),while the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly increased(P<0.001,P<0.05),the number of c-Fos positive cells significantly increased(P<0.001).Compared with the model group,the PWT and PWL of the tuina group significantly increased on the 10th and 17th day after modeling(P<0.05,P<0.01,P<0.001),and the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly decreased(P<0.001,P<0.01,P<0.05),and the number of c-Fos positive cells significantly reduced(P<0.001).Conclusion Tuina may inhibit the activation of microglia by regulating P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression in spinal dorsal horn,reduce neuronal excitability,and exert analgesic effects on lumbar disc herniation.