Research method for applying TLR4 to curing acute kidney injury caused by sepsis
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A technology for acute kidney injury and sepsis, which is applied in the medical field and can solve problems such as delay
Inactive Publication Date: 2018-09-11
邦世(苏州)生物医药科技有限公司
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However, at present, there are great controversies on what indicators should be used as the predictors of AKI in critically ill patients, especially those with sepsis, the signs of early AKI, a
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specific Embodiment approach 1
[0078] see as figure 1 Shown, the technical scheme that this specific embodiment adopts is: it carries out research analysis from cell experiment approach, and its research method is as follows:
[0079] 1. Establishment of septic AKI model in mice:
[0080] 1.1. A single intraperitoneal injection of LPS (10mL / kg) was used to prepare the model of septic AKI; 40 Balb / c mice were divided into two groups according to the random number table method, namely the control group and the model group, 20 in each group ; Normal saline (NS) control group: intraperitoneal administration of the same amount of normal saline; model group: mice were intraperitoneally injected with 10 mL / kg of LPS; fasted for 12 hours before modeling, free to drink, and free to drink and eat after modeling;
[0081] 2. Determination of animal modeling:
[0082] 2.1. The performance of the mice after stimulation: the control group was flexible in activities, free to drink water, erect hair, flexible in activiti...
specific Embodiment approach 2
[0134] see figure 2 , the technical scheme that this specific embodiment adopts is: it carries out research analysis from cell experiment approach, and its research method is as follows:
[0135] 1. Construct a mouse model of septic AKI, and transfect the lentiviral expression vector containing VEGF overexpression / silencing into septic AKI mice:
[0136] Balb / c mice were divided into 5 groups according to the random number table method, including 10 blank control groups; 4 groups of septic AKI group, 10 mice in each group, respectively: septic AKI control group, without Any intervention; Ad-GFP group; Ad-VEGF group; Ad-VEGF-ShRNA group. The model of septic AKI mice was the same as before, starting from the 10th day, Ad-VEGF and Ad-VEGF-ShRNA group mice were intraperitoneally injected with recombinant lentivirus containing Ad-VEGF and Ad-VEGF-ShRNA, Ad-GFP Group mice were transfected with a lentiviral vector containing only the GFP genome. The transfection method was once a...
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Abstract
The invention provides a research method for applying TLR4 to curing acute kidney injury caused by sepsis, and relates to the technical field of medical treatment. The method comprises the steps of mouse septic AKI model building; animal modelling judgement; specimen collection; normal and septic AKI mouse kidney cell primary culture; TLR4 determination by means of immunohistochemistry (SP method)and semi-quantitation; Real-time PCR detection of expression change of VEGF, sFLT-1, TNF-alpha and TLR4mRNA of septic AKI mouse primary kidney cells; detection of protein level expression change of VEGF, sFLT-1, TNF-alpha and TLR4 of septic AKI mouse primary kidney cells through a Western-blot method; constructing VEGF gene eukaryotic expression plasmids and optimal silence mouse VEGF gene ShRNAexpression plasmids; experimental grouping; TLR4 cell treatment; index detection; verifying TLR4 target regulation on VEGR expression; searching for protein in interaction with the VEGF; discussing molecule mechanisms which is possibly involved in TLRs signal path adjustment of the VEGF. The method speculates that the VEGF and the TLR4 signal path are in interaction for regulating the acute kidneyinjury caused by sepsis and play a part in protection.
Description
technical field [0001] The invention relates to the field of medical technology, in particular to a research method for applying TLR4 to the treatment of sepsis acute kidney injury. Background technique [0002] Sepsis is the leading cause of death in critically ill patients, and its incidence is increasing day by day. Studies have shown that the incidence of severe sepsis during hospitalization has almost doubled in the past decade, which has also greatly increased its mortality rate. increased risk. Sepsis can lead to multiorgan failure, such as acute kidney injury. For critically ill patients, sepsis is currently the leading cause of AKI. AKI often complicates sepsis or infectious disease. So-called sepsis-AKI is independently associated with increased risk of death and prolonged hospital stay. Correlation. Although some progress has been made in active anti-infection treatment, vital organ function support and resuscitation methods, the mortality rate of sepsis AKI is...
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