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ObjectiveTo investigate the analgesic effect and mechanism of Osteoking （OK） on nerve compression in lumbar disc herniation. MethodThe rat model of chronic compression of dorsal root ganglion （CCD） was established to simulate clinical lumbar disc herniation. The CCD rats were randomly divided into model group， low， medium， and high dose OK groups （1.31， 2.63， 5.25 mL·kg-1·d-1）， and pregabalin group （5 mg·kg-1）， with eight rats in each group. Another eight SD rats were taken as the blank group， and the same volume of normal saline was given by gavage. Behavioral tests， side effect evaluation， network analysis， Western blot， immunofluorescence， and antagonist application were used to explore the effect. ResultCompared with the blank group， the mechanical hyperalgesia threshold， thermal hyperalgesia threshold， and the expression of inflammatory factors in the spinal dorsal horn of the model group are significantly increased （P<0.01）， and the related indicators of the affected foot footprints are significantly down-regulated （P<0.01）. The expression of signal transducer and activator of transcription 3 （STAT3）， vascular endothelial growth factor A （VEGFA）， and phosphorylated extracellular regulated protein kinase （p-ERK） in microglia in the spinal dorsal horn is significantly increased in the model group （P<0.01）. Compared with the model group， low， medium， and high dose OK groups can increase the mechanical hyperalgesia and thermal hyperalgesia thresholds of CCD rats （P<0.05， P<0.01） in a dose-dependent manner， improve the gait of CCD rats （P<0.05， P<0.01）， and reduce the expression of inflammatory factors in the spinal dorsal horn （P<0.05， P<0.01）. The expression of STAT3， VEGFA， and p-ERK in the spinal dorsal horn microglia of CCD rats is significantly decreased （P<0.05， P<0.01）， and the acetic acid-induced nociceptive response in rats is effectively reduced （P<0.05， P<0.01）. In addition， there is no tolerance. The results of the body mass test， organ index， forced swimming， and rotation show that OK has no obvious toxic or side effects. Further antagonist experiments show that MRS1523 and RS127445 can reverse the transient analgesic effect of OK compared with the high dose OK group （P<0.01）. ConclusionOK has a good analgesic effect on the CCD model without obvious toxic side effects， and its mechanism may be related to the activation of ADORA3 and HTR2B and the inhibition of STAT3， VEGFA， p-ERK， and other elements in microglia.
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@# Objective：To explore the targeting relationship between miR-377-5p and hypoxia inducible factor-1 (HIF-1α), and investigate the regulatory effect of miR-377-5p on proliferation, invasion and epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells through vascular endothelial growth factor (VEGF) signaling pathway. Methods： ：The expression of miR-377-5p in 35 pairs of human HCC tissues and para-cancerous tissues was detected by qPCR. Then, HepG2 cells were divided into control group, mimic-NC group and miR-377-5pmimicgroup.qPCRwasusedto detect the transfection efficiency; the effects of miR-377-5p over-expression on proliferation and invasion of HepG2 cells were examined by EdU staining and Transwell assay, respectively; and the effect of miR-377-5p over-expression on the expressions of proliferation-related protein Ki-67, proliferating cell nuclear antigen (PCNA) and epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin) were detected by Western blotting (WB); the effect of miR-377-5p over-expression on the expression of hypoxia inducible factor-1α (HIF-1α) in HepG2 cells was detected by qPCR and WB; and the targeting relationship between miR-377-5p and HIF-1α gene was determined by Luciferase reporter gene assay. Results: The expression of miR-377-5p in HCC tissues was significantly lower than that in para-cancerous tissues (P<0.01). Compared with the control group, the expression of miR-377-5p in HepG2 cells of miR-377-5p mimic group elevated significantly, and the proliferation, invasion and the expression of N-caderin proteins decreased,significantly (all P<0.01), while the expression of E-caderin increased significantly (P<0.01). At the same time, the mRNA and protein expressions of HIF-1α in miR-377-5p mimic group decreased significantly (P<0.01 or P<0.05). miR-377-5p targetedly inhibited the expression of HIF-1α gene and suppressed the activation of VEGF pathway (all P<0.05). Conclusion: miR-377-5p inhibits the proliferation, invasion and EMT of HepG2 cells via targetedly inhibiting HIF-1α expression and suppressing the activation of VEGF signaling pathway.
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0.05);HIF-1?positive rate of control groups was higher than that in normal group and treatment group with significant difference among them(P0.05);VEGF positive rate in control groups was higher than that in normal group and treatment group with significant difference among them(P
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0.05).The transfected NSCs were confirmed to have the latent ability of multi-directional differentiation.Conclusion:NSCs can express HIF-1? and GFP steadily after transfected with the recombinant adenovirus and the bionomics is normal.
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Objective:To investigate the influence of Bifidobacterium adolescence on the expression of HIF-1?and VEGF in rabbit VX2 tumor.Methods:Rabbit VX2 tumor models were established and were divided into two groups(n= 10).Animals in the experimental group were injected with Bifidobacterium adolescence and those in the control group received normal saline via ear vein.Animals were sacrificed and the liver,kidney,spleen,heart,lung and tumor tissues were removed,cultured and Gram stained.The changes of tumor tissue were observed by H-E staining;immunohistochemistry was used to detect the expression of HIF-1?and VEGF in tumor tissues.Results:Bifidobacterium adolescence could target the tumor tissue.H-E staining showed the necrotic area was obviously enlarged after treatment with Bifidobacterium. Immunohistochemistry demonstrated that the positive rates of HIF-1?and VEGF in the experimental group were significantly lower than those of the control group(P
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0.05).In spinal cord tissues of injury groups,the level of HIF-1? mRNA began to elevate at 3h after injury and reached the maximize on 3d,then it began to decline and returned to the basal level on 14d;the HIF-1? expression showed a time-dependent difference as followed: It began to increase at 3h after injury,peaked on 1d,then gradually decreased,and recovered to the normal level on 14d after injury.Conclusion The result suggested that hypoxia circumstance can enhance the stability of HIF-1? and the transcription of HIF-1? mRNA,and the time-dependent expression of HIF-1? and HIF-1? mRNA can be used to help the diagnosis of SCI and estimation of injury time.
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Objective:To study the effect of Hypoxia-inducible factor-1 alpha(HIF-1?)on the radiosensitivity of rat cervical carcinoma and discuss the possible mechanism.Methods:The transplantation tumor model of cervical carcinoma was established. 77 rats were randomly divided into five groups:control group,the large fraction irradiation group,the small fraction irradiation group,the large fraction irradiation combining HBO group and the low dose irradiation combining HBO group.Expressions of HIF-1? and bcl-2 in transplantation tumor after treatment were detected by immunohistochemical staining,and the tumor inhibitory rate by the change of tumor volume.The correlation between expressions of HIF-1 ? and bcl-2(and tumor inhibitory rate)was also observed.Results:Expression of HIF-1 ? and carcinoma growth inhibitory rate showed a reverse correlation(r=-5.147,P
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Objective:To investigate the effect of hypoxia on vascular endothelial growth factor expression in breast carcinoma cell line(MD-MBA-435S) and its mechanism.Methods:Hypoxia culture model in vitro was established to test the expression of VEGF and HIF-1? and their correlation in MD-MBA-435S cell line.After cultured in hypoxia circumstance for a given period of time(4 hours,8 hours,12 hours,16 hours,20 hours),VEGF expression was tested in mRNA level and protein level by RT-PCR and Western blot analysis,HIF-1 ? expression tested in protein level.Cell viability in twenty hours was tested by trypan blue exclusion assay.Results:Cell survival was affected a little within 20 hours under hypoxia.Two isoforms of VEGFmRNA,VEGF-121 and VEGF165,were identified by RT-PCR,and their expressions increased after treatment of hypoxia for 4 hours.VEGF and HIF-1 ? protein expressions increased equally with VEGFmRNA confirmed by immunocytochemistry and Western blot and its positive correlation was found by Pearson product-moment correlation analysis.Conclusion:VEGF gene and protein can be induced remarkably by hypoxia in breast carcinoma cell line,which is regulated mostly by transcription factor HIF-1 ?.This mechanism may be involved in the malignant proliferation and transformation of breast neoplasm.