| Background Hypoxic-ischemic encephalopathy(HIE)can cause irreversible brain damage,during the disease brine can release a lot of inflammatory cytokines,which are important factor in the promotion of secondary brain damage occurs,however,in addition to hypothermia outside so far no satisfactory treatment measures,so finding a safe and effective treatment is very important.Study confirmed administered fish oil rich in omega-3 fat emulsion supplement can significantly improve ischemic brain disease progression and prognosis,and focus on adding DHA omega-3 fish oil fat emulsion is a brain hormone synthesis of neuroprotective D1(NeuroprotectinD1,NPD1)of important sources.HIE required when parenteral nutrition therapy to meet the energy needs of children,and therefore able to supply energy but at the same time curb the development of the disease,and explore the mechanism of anti-inflammatory ?-3 fish oil fat emulsion became the starting point of this experiment.Objective To discuss omega-3 fish oil fat emulsion the potential protective mechanism to Seven-day-old rats after hypoxic-ischemic brain damage.Methods 168 Seven-day-old SD rats were randomly divided into four groups(n=42),sham group,omega-3 fish oil fat emulsion group,20% ordinary fat milk group and model group.Neonatal hypoxic-ischemic encephalopathy model was produced by the method of Rice.Briefly,P7 rats were anesthetized with diethyl ether,and the left common carotid artery was ligated using surgical silk,followed by exposure to 8% oxygen for 2 h in a heated box at 37°C.Omega-3 fish oil fat emulsion,20% ordinary fat milk and model groups were handled with method of RICE.Sham group only exposed the left carotid artery.Rats were sacrificed at P8,P10 and P14 after the surgery.Hippocampus wereremoved for Real-time PCR and Western-Blot test,to detective expression of TLR4 and NF-?B mRNA and protein.TUNEL staining comparison between different groups to observe the number of cell apoptosis.Results1.Model evaluation: Omega-3 fish oil fat emulsion,20% ordinary fat milk and model groups after the surgery may occur standing firm and whirling towards left,and sham group has not these phenomenon,TTC staining shows that Omega-3 fish oil fat emulsion,20% ordinary fat milk and model groups brain tissues with large area of cortical infarct,but the sham group rat brain slices a uniform red,without white infarction area.It is suggesting that the model was successfully established.(Figure 1)2.Brain tissue pathological: HE staining of hippocampus CA1 region in 3 d shows that sham group brain tissues maintain normal structure.Omega-3 fish oil fat emulsion,20% ordinary fat milk and model groups were seen edema,disordered pyramidal cell and accumulated chromatin,etc.20% ordinary fat milk and model groups pathology represent much more severe than omega-3 fish oil fat emulsion group.(Figure 2)3.Real-time PCR results shows that omega-3 fish oil fat emulsion can reduce the mRNA expression levels of TLR4 and NF-?B: TLR4 and NF-?B mRNA expression levels in model group was higher than sham group in 1 d(P = 0.00,P <0.05);TLR4 and NF-?B expression levels of mRNA in omega-3 fish oil fat emulsion group [(4.89 ± 0.51),(1.15 ±0.10)] lower than 20% ordinary fat milk group [(17.58 ± 2.50),(2.56 ± 0.10)] and model group [(15.94 ± 2.52),(2.34 ± 0.11)] in 3d(P = 0.00,P <0.05),compared with sham group[(6.30 ± 1.52),(1.32 ± 0.10)] there is no significant difference(P = 0.376,0.21,P>0.05);TLR4 and NF-?B mRNA expression levels in group omega-3 fish oil fat emulsion group lower than that in 20% ordinary fat milk group(P = 0.03,0.04,P <0.05)and model group in 7 d(P = 0.00,P <0.05),compared with sham group there is no significant difference(P = 0.23,0.08,P> 0.05).(Figure 3,Table4)4.Western-bloting results shows that omega-3 fish oil fat emulsion can reduce theprotein expression levels of TLR4 and NF-?B: Western blotting was used to test the expression levels of TLR4 and NF-?B for four groups,GAPDH as internal control,the results shows that TLR4 and NF-?B protein expression levels in group model group was higher than sham group in 1 d(P = 0.00,P <0.05);TLR4 and NF-?B expression levels of protein in omega-3 fish oil fat emulsion group [(9.30 ± 1.53),(1.44 ± 0.14)] lower than20% ordinary fat milk group [(20.13 ± 1.00),(2.82 ± 0.09)] and model group [(26.21 ±3.00),(4.51 ± 0.36)] in 3 d(P = 0.00,P <0.05),compared with sham group [(5.32 ± 1.06),(2.42 ± 0.14)] there is no significant difference(P = 0.2,0.17,P> 0.05);TLR4 and NF-?B protein expression levels in omega-3 fish oil fat emulsion group lower than that in 20%ordinary fat milk group(P = 0.00,0.02,P <0.05)and model group in 7 d(P = 0.00,P <0.05),compared with sham group there is no significant difference(P = 0.21,0.33,P>0.05).(Figure 4,5,Table 5)5.The apoptotic cell numbers of brain tissues in 3 d: omega-3 fish oil fat emulsion group(13.67 ± 2.52)lower than 20% ordinary fat milk group(27.67 ± 2.52)and model group(41.00 ± 3.61)(P=0.00,P <0.05),compared with the sham group(6.00 ± 2.00),the difference was not statistically significant(P=0.09,P>0.05).(Figure 6)ConclusionOmega-3 fish oil fat emulsion play a important role in protecting neonatal rats from hypoxic-ischemic damage.The mechanisms were likely to reduce TLR4,NF-?B and cell apoptosis levels. |
PDF Full Text Request