ABSTRACT
Objective To evaluate the effect of different doses of Astragalus membranaceus on the levels of vascular endothelial growth factor ( VEGF) and basic fibroblast growth factor ( bFGF) in serum and lung tissues of rats with pulmonary embolism. Methods Seventy-six clean-grade healthy male Sprague-Dawley rats, aged 8-9 weeks, weighing 140-170 g, were assigned to control group ( group C, n=11) and experimental group ( group E, n=65) by a random number table method. The rats with pulmonary em-bolism in group E were further divided into 4 subgroups using a random number table method: pulmonary embolism group (group P), low-dose Astragalus membranaceus group (group H1), median-dose Astraga-lus membranaceus group ( group H2 ) and high-dose Astragalus membranaceus group ( group H3 ) . The model of pulmonary embolism was established by injecting autologous blood clots into the right jugular vein. At 1 h and 1, 2, 3, 4, 5 and 6 days after successful establishment of the model, Astragalus membrana-ceus 20, 40 and 60 g∕kg were injected intraperitoneally in H1-3 groups, respectively, while the equal vol-ume of normal saline was given instead in group P. The chest was opened after anesthesia on day 7, and blood samples were collected from cardiac chambers for determination of concentrations of serum VEGF and bFGF by enzyme-linked immunosorbent assay. The pulmonary specimens were obtained from the upper lobe of right lungs for determination of the expression of VEGF and bFGF mRNA ( using real-time polymerase chain reaction) . Results Compared with group C, the concentrations of serum VEGF and bFGF were sig-nificantly increased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in the other four groups (P<0. 05). Compared with group P, the serum bFGF concentration was significantly in-creased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in H1-3 groups ( P<0. 05) . Compared with group H1, the serum bFGF concentration was significantly increased, the ex-pression of VEGF mRNA and bFGF mRNA in lung tissues was up-regulated in H2 and H3 groups ( P<0. 05) . Compared with group H2, the expression of VEGF and bFGF mRNA in lung tissues was significant-ly up-regulated in group H3 ( P<0. 05 ) . Conclusion Astragalus membranaceus can up-regulate the ex-pression of VEGF and bFGF in lung tissues in a dose-dependent manner, thus improving pulmonary embol-ism in rats.
ABSTRACT
Objective To investigate the signaling pathway and the key signal molecules of protein kinase (RIPK)3 in SH-SY5Y cells. Methods SH-SY5Y cells were transfected with RIPK3 expression plasmid vector to upregulate intracellular RIPK3, while the SH-SY5Y cells were transfected with empty vector plasmid, which was considered as control group. Western blot assay was used to check the expression of exogenous RIPK3 in cells. The proliferation rate of SH-SY5Y cells was determined by MTT assay at designated time to detect exogenous RIPK3 activity. Whole transcriptome sequencing (RNAseq) was used to detect the transcription of genes. Whole-transcriptomic gene transcription was measured by following Ingenuity Pathway Analysis (IPA) to obtain downstream signaling pathways and the key molecule, which were partly confirmed by following droplet digital PCR (ddPCR). Results Exogenous RIPK3 showed biological activity in SH-SY5Y, which inhibited the proliferation of cells. IPA showed that znic finger protein 36 (ZFP36) was significantly up-regulated as compared with that of the control group. The tran?scription levels of ZFP36 downstream genes such as tumor necrosis factor (TNF), brain derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF) and mRNA-decapping enzyme 2 (DCP2) were affected at the same time. Conclusion Within the limitations of this study, it seems that RIPK3 is notable for the development, inflammation and tumorigenesis of the nervous system as an independent regulator of ZFP36 gene and downstream effectors.
ABSTRACT
Objective To observe the changes of ischemic preconditioning on the expression of hypoxia inducible fac?tor (HIF)-1αand vascular endothelial growth factor (VEGF) in ischemia hippocampus CA1 region after focal cerebral isch?emia/reperfusion (I/R) in rats, and the mechanisms of brain protection from brain ischemia preconditioning (BIP) thereof. Methods The male SD rats were randomly divided into three groups:sham operation (SO) group,middle cerebral artery oc?clusion (MCAO) group and brain ischemia preconditioning (BIP) group. The MCAO group and BIP group were further divid?ed into six subgroups according to perfusion time after I/R including 2 h, 6 h, 12 h, 24 h, 48 h and 72 h. The ischemia pre?conditioning model rats were established. Immunohistochemistry and Western blot assay were used to observe the expres?sions of HIF-1αand VEGF in ischemia hippocampal CA1 region. Results Neurological function deficit was not observed in SO group. Compared with MCAO group, there was a lower neurological function deficit score in BIP group. In MCAO group and BIP group, the expressions of HIF-1αand VEGF positive cells and protein increased at 2 h after I/R, then gradu?ally increased from 6 h to12 h and reached the maximum level at 24 h, then gradually decreased. The levels were still higher at 72 h than those of SO group. The number of HIF-1αand VEGF positive cells and protein were significantly increased in MCAO group and BIP group than that of SO group (P<0.05). The number of HIF-1αpositive cells was higher in BIP group than that in MCAO group except 2 h and 6 h reperfusion groups. The expression of VEGF positive cells, HIF-1αand VEGF protein were significantly higher in BIP group than those in MCAO group at different time points (P < 0.05). Conclusion Ischemic preconditioning plays a protective role in brain, which may be related to up-regulation of HIF-1αand VEGF.