| The current version does not support copying Cyrillic text to the Clipboard. You should activate the programexpression, and the percentage of positive cells and ratios of light transmittance of cytoplasma and cell nucleus6. Adopting the method of RT-PCR to observe the effect of different concentrationsof different drugs on the expression of ICAM-1 mRNA and VCAM-1 mRNA of microvascular endothelial cells after hypoxia/reoxygenation injury.7. Adopting SPSS11.0 to carry out statistics and variance analysis.Result1. Some capillary segments can be found, cells growing monoptychially and mixing together in the course of cells extraction. Detected by immunofluorescence method , the correlated antigen of VIII factor is positive. Microvillus, fenestration, as well as tight cell conjunction can be observed through scanning electron microscope. All these facts prove that the cells extracted are cerebral microvascular endothelial cell of rat.2. The rats MVEC were cultured in vitro. The MVEC monolayers were exposed to hypoxia for 4h, reoxygenate for 12h. The hypoxia/reoxygenation model is successfully replicated.3. Synthesizing the result of MTT in normal and hypoxia/reoxygenation injury condition, it is determined in the experiment that the best final concentration of the drugs are jasminoidin 0.128umol/ml, 0. 064 ii mol/ml and 0. 032 u mol/ml, baicalin 0.028 u mol/ml, 0.014 u mol/ml and 0. 007 u mol/ml, berberine 0.024u mol/ml, 0. 012u mol/ml and 0. 006u mol/ml, and Qing Kai Ling 0.025%.4. Hypoxia/reoxygenation injury can significantly raise the amount of protein expression of microvascular endothelial molecules NF- k , promote the movement of NF- k from cytoplasm to nucleus, and enhance the activity of nucleus transcription. Jasminoidin, baicalin and berberine can significantly inhibit the activity and protein expression of NF-kB, inhibit the movement of NF- k. from cytoplasm to nucleus and the activity of nucleus transcription.5. Hypoxia/reoxygenation injury can induce the high expression of ICAM-1 and VCAM-1 on the surface of microvascular endothelial molecules, with the expression of ICAM-1 stronger than that of VCAM-1. The experimental drugs can inhibit the expression of the protein of ICAM-1 and VCAM-1. 0.128y mol/ml jasminoidin and 0.007 p mol/ml baicalin can significantly inhibit the expression of VCAM-lmRNA. They can reduce the expression of VCAM-lmRNA but have no statistics significance. While drugs of other groups can significantly inhibit ICAM-lmRNA and VCAM-lmRNA expression . It may relate that the expression of mRNA precede proteinum andICAM-1 precede VCAM-1 and instability of baicalin solute .6. The inhibiting effect of Jasminoidin, baicalin and berberine on the expression of ICAM-1, VCAM-1mRNA promote gradually from lower concentration to higher one.ConclusionThe Hypoxia/Reoxygenation model can imitate ischemic-reperfusion injury fairly good. Hypoxia/Reoxygenation injury can significantly enhance the activity of NF- k B , promote the movement of NF- k B nucleus, enhance the activity of nucleus transcription of NF-kB, and induce the high expression of ICAM-1 and VCAM-1. The effective components of Huang Lian Jie Du Tang and Qing Kai Ling can significantly inhibit the activity and protein expression of NF- k B, reduce it' s movement from kytoplasm to nucleus , inhibit the activity of nucleus transcription of NF-k B ,and reduce the high expression of ICAM-1 and VCAM-1. All of these indicate that Jasminoidin, baicalin, berberine and Qing Kai Ling can inhibit the expression of ICAM-lmRNA and VCAM-lmRNA, reduce the inflammatory reaction due to hypoxia/reoxygenation injury, thus antagonize the cranial nerve injury due to inflammatory reaction and brain injury due to hypoxia/reoxygenation. The mechanism maybe related to the inhibition of the activity and protein expression of NF- k B and reduction of the movement of NF- k B from kytoplasm to nucleus. The study provide a new idea and a new way for studying the mechanism of herbs compounds and monomers treating brain injury caused by hypoxia/reoxygenation. |
PDF Full Text Request