ABSTRACT
Objective:To evaluate the effects of different concentrations of ropivacaine on the growth and migration of lung cancer cells.Methods:Human lung adenocarcinoma cell strain A549 cells and human lung squamous cell strain H520 cells were divided into 4 groups ( n=24 each) using a random number table method: control group (group C) and different concentrations of ropivacaine groups (Ⅰ-Ⅲ groups). Cells were commonly cultured in group C. Ropivacaine 3, 5 and 7 mmol/L were added and then the cells were cultured in Ⅰ-Ⅲ groups, respectively. The cell survival rate was determined using the CCK-8 method at 24, 48 and 72 h of treatment (T 1-3). The cell cycle and apoptosis were detected at T 1 using flow cytometry. The expression of Cyclin D1, cyclin-dependent kinase 4 (CDK4), cleaved poly (ADP-ribose) polymerase-1 (PARP-1) and cleaved caspase-3 was detected using Western blot. Wound healing assay was used to measure cell migration distance. The activities of RhoA and Rac1 were detected by microplate spectrophotometry. Results:The cell viability of A549 and H520 cells sequentially decreased at T 1-3, the proportion of G0/G1 phase and apoptosis sequentially increased, the expression of Cyclin D1 and CDK4 was down-regulated sequentially at T 1, the expression of cleaved PARP-1 and cleaved caspase-3 was up-regulated sequentially, and the cell migration distance, RhoA, and Rac1 activity decreased sequentially in C, Ⅰ, Ⅱ and Ⅲ groups ( P<0.05). Conclusions:Ropivacaine can inhibit the growth and migration ability of lung cancer cells in a concentration-dependent manner, which is related to induction of cell cycle arrest and apoptosis.
ABSTRACT
Objective:To evaluate the role of Nod-like receptor family pyrin domain-containing 3 (NLRP3) in the dorsal root ganglion (DRG) in persistent postoperative pain in rats.Methods:Forty-eight male Sprague-Dawley rats, in which IT catheters were successfully implanted, weighing 200-250 g, aged 2-3 months, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (S group), persistent postoperative pain group (P group), persistent postoperative pain+ NLRP3-siRNA group (P+ siRNA group) and persistent postoperative pain+ NLRP3-scrRNA group (P+ scrRNA group). A persistent postoperative pain model was established by skin/muscle incision and retraction (SMIR) in anesthetized animals.Normal saline 10 μl were intrathecally injected in S group and P group, NLRP3-siRNA 10 μl and NLRP3-scrRNA 10 μl were intrathecally injected in CP+ siRNA group and CP+ scrRNA group, respectively, at 3 days before operation.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before operation (T 0) and 1, 5, 10, 15 and 20 days after operation (T 1-5). Six rats in each group were randomly selected and sacrificed at T 3, and the L 4-5 DRGs on the operated side were harvested for determination of the expression of NLRP3 and caspase-1 (by Western blot), NLRP3 mRNA expression (by real-time polymerase chain reaction) and content of interleukin-1beta (IL-1β) (by enzyme-linked immunosorbent assay). Results:Compared with group S, the MWT was significantly decreased at T 2-5, the expression of NLRP3 protein and mRNA in DRGs was up-regulated, and the content of IL-1β was increased in group P ( P<0.05). Compared with group P, the MWT was significantly increased at T 2-5, the expression of NLRP3 protein and mRNA and caspase-1 was down-regulated, and the content IL-1β was decreased in group P+ siRNA ( P<0.05), and no significant change was found in the parameters mentioned above in group P+ scrRNA ( P>0.05). Conclusion:NLRP3 in DRGs is involved in the development of persistent postoperative pain, and the mechanism may be related to the development of NLPR3 inflammasomes which further induces peripheral neuroinflammatory response in rats.