ABSTRACT
Objective:To observe the effects of the Chinese medicine prescription Xiao-Cheng-Qi decoction (XCQD) on acute brain edema and inflammatory factors in rats with severe traumatic brain injury (sTBI).Methods:A total of 108 male Sprague-Dawley (SD) rats were divided into control group, sham operation group, sTBI model group, and XCQD low, medium, high dose groups by random number table method, with 18 rats in each group. sTBI rat model was prepared according to the modified Freeney method. At 6 hours after injury, the XCQD low, medium, and high dose groups were given XCQD 1.80, 2.78, and 4.59 g/kg by gavage, respectively, and the other three groups were given the same amount of normal saline, once a day for 3 days. After 3 days of injury, rats in each group were sacrificed after the modified neurologic severity score (mNSS) assessed. Pathological changes of brain tissue were observed under light microscope after hematoxylin eosin (HE) staining, water content of brain tissue was measured by dry-wet specific gravity method, and the expressions of aquaporin 4 (AQP4), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue were detected by Western blotting. Serum TNF-α and IL-1β levels were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the normal group, the mNSS score of rats increased significantly, the structure of brain tissue was disordered, and pathological changes appeared such as inflammation, edema, pyknosis of nerve nuclei, water content, the protein expressions of AQP4, TNF-α and IL-1β in brain tissue, and the contents of TNF-α, IL-1β in serum were significantly increased. After XCQD intervention, the above indexes were significantly improved. Compared with sTBI model group, the mNSS score of XCQD medium and high dose groups significantly decreased (6.94±1.16, 6.88±1.02 vs. 8.61±1.09, both P < 0.05), and the pathological changes such as brain edema and inflammation were alleviated. Brain tissue water content, AQP4 protein expression and contents of serum TNF-α, IL-1β in XCQD low, medium, and high dose groups significantly decreased compared with sTBI model group [brain tissue water content: (78.25±0.71)%, (77.62±0.44)%, (76.70±0.74)% vs. (80.08±0.66)%; the expression of brain AQP4 protein (AQP4/β-actin): 0.86±0.13, 0.84±0.22, 0.65±0.13 vs. 1.08±0.14; serum TNF-α (ng/L): 106.34±15.07, 95.75±17.26, 89.00±17.36 vs. 141.96±29.47; serum IL-1β (ng/L): 90.41±12.88, 72.82±13.51, 71.32±16.79 vs. 128.57±22.56, respectively, all P < 0.05]. The protein expressions of TNF-α,IL-1β in brain tissue of XCQD medium and high dose groups also significantly decreased compared with sTBI model group [TNF-α (TNF-α/β-actin): 0.90±0.24, 0.79±0.35 vs. 1.17±0.15; IL-1β (IL-1β/β-actin): 0.91±0.21, 0.68±0.28 vs. 1.23±0.08, respectively, all P < 0.05]. Brain tissue water content, the expression of brain AQP4 protein, the levels of brain tissue and serum IL-1β in XCQD high dose group improved more significant than those of XCQD low dose group. Conclusions:XCQD can alleviate the acute brain edema in sTBI rats, and it is dose-dependent. The mechanism may be relevant to reduce the secondary inflammatory response of sTBI by inhibiting the expression of inflammatory factors TNF-α and IL-1β.
ABSTRACT
Objective To investigate the effect of LMO2 gene,which is overexpressed in prostate peripheral zone than transitional zone,on proliferation and invasion of prostate cancer PC-3 cell line or normal prostate epithelial BPH-1 cell line.Methods Lentivirus overexpression system was used to establish the LMO2 protein overexpressed prostate WPMY-1 stromal cell line.The LMO2 mRNA and protein expression level of those cells was evaluated by qRT-PCR and western blot.The PC-3 or BPH-1 cells was cocultured with prostate stromal cells and the in vitro proliferation and invasion activity were detected by Cell Counting Kit-8 (CCK-8),EdU and Matrigel invasive assays.Results The stable LMO2 overexpressed prostate stromal cells WPMY-1-LMO2 was successfully established.The results of CCK-8,EdU experiments indicated the proliferation and invasion activity of PC-3 or BPH-1 cells were both enhanced through cocultured with WPMY-1-LMO2 cells.The proportion of EdU positive cells of PC-3 cultured in WPMY-1-LMO2 supernatant was (42.67 ± 6.03) ％,and the difference was significant compared with the control group (29.33 ± 3.51) ％ (P ＜ 0.05).The BPH-1 culturcd in the supernatant was (35.00 ± 2.52) ％,aud the difference was significant compared with the control group (23.33 ± 2.52) ％ (P ＜ 0.05).The number of invaded PC-3 or BPH-1 cells after co-cultured with WPMY-1-LMO2 cells was 38 ± 5 and 43 ± 3,respectively,which was significantly different compared with the control group (P ＜ 0.05).Conclusions Tbe LMO2 overexpressed prostate stromal cells could induce proliferation and invasion of PC-3 or BPH-1 cells.The propensity of benign prostatic hyperplasia and prostate cancer at different zones may probably be related to distinctive gene expression between peripheral zone and transitional zone derived stromal cells.