1.The effect of recombinant human erythropoietin on the function of brain mitochondria after brain injury
Zhian ZHU ; Dongfu FENG ; Yanbin MA ; Hui WU ; Xiaoyan SHEN ; Yingfan XIONG ; Fangfang WU ; Shenghua CHU
Chinese Journal of Emergency Medicine 2008;17(5):503-506
Objective To investigate the therapeutic effects of recombinant human erythropoietin(rhEPO)on brain mitochondrial energy metabolism and mitachondrial respiratory functionin after brain injury in rats.Method A total of 63 Sprague-Dawley rats were divided randomly into three groups:the rhEPO treated group(n =28),the control group(n=28),the shanl group(n=7).The models of contusion of brain caused by freefalling were set up in rhEPO treated group(n=28).The recombinant human erythropoietin was intraperitoneally injected in dose of 10 U/g immediately after brain injury and it was repeated every 10 hours in rhEPO group treated.The same models of contused brain were made without rhEPO treatment as control group(n=28).In control group,the same volume of normal saline was used in replacemem of rhEPO.Aburr hole was made on the skull of the sham group(n=7),but the brain tissue was not wounded.The mitochondria were isolated at 6 h,12 h,24h,48 h after trealment,respectively.The activity of ATPase and SOD,the content MDA and brain mitochondrial respiratory function were measured by biochemical technique.The data were analyzed with the F-test and t-test.Results The activity of ATPase(P<0.05),SOD(P<0.01)and brain mitochondrial respiratory function(P <0.05)were increased.and the levd of MDA in brain mitochondria was reduced markedly in rats treated with rhEPO.Conclusions Treatment with rhEPO can alleviates the secondary brain injury by affecting mitochondrial function.
2.Effects of Bushen Huoxue Formula (补肾活血方) on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice
Xiaorong QI ; Feiran HAO ; Xianglin TANG ; Fagen LI ; Yujia WANG ; Liang WANG ; Yingfan SHEN ; Minghui YANG ; Min LI
Journal of Traditional Chinese Medicine 2024;65(10):1038-1045
ObjectiveTo explore the possible mechanism of Bushen Huoxue Formula (补肾活血方, BHF) in the treatment of Parkinson's disease (PD) from the the perspective of intestinal flora. MethodsSeventy-two male C57/BL6J mice were randomly divided into blank group, model group, Madopar group and low-, medium- and high-dose BHF groups, with 12 mice in each group. The mice in the blank group were intraperitoneally injected with 10 ml/kg of normal saline, and those in the other groups were intraperitoneally injected with 30 mg/kg of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) at a concentration of 3 mg/ml to induce PD mice model, both once a day for 7 consecutive days. After successful modeling, the low-, medium-, and high-dose BHF groups were given 7.5, 15, and 30 g/(kg·d) of BHF by gavage, respectively, while the Madopar group was given 112.5 mg/(kg ·d) of Domedopar tablets by gavage, and the blank group and the model group were given 15 ml/(kg·d) of distilled water, all once a day for 14 consecutive days. The rod climbing test, rotating rod test, grip strength test and weight-bearing swimming test were used to evaluate the behavioral indicators of mice. Western blotting was used to measure the protein expression levels of Toll-like receptor (TLR)/nuclear factor kappa B (NF-κB) pathway inflammatory factors in the mouse ileum, including Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), NF-κB, tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 17 (IL- 17). 16S rRNA high-throughput sequencing was used to analyze changes in mouse intestinal flora. ResultsCompared to those in the blank group, the mice in the model group had longer bottoming time when climbing the pole, reduced grip strength, shortened rotary pole duration and swimming duration, and increased protein expression levels of TLR2, TLR4, NF-κB, TNF-α, and IL-6 in the ileal tissue (P<0.01). Compared to the model group, the Madopar group and the low-, medium- and high-dose BHF groups had shortened bottoming time of the climbing pole and increased grip strength; the Madopar group and the high-dose BHF group had prolonged rotary pole duration, and reduced protein expressions of TLR2, TLR4, NF-κB, TNF-α, IL-6, and IL-17 levels; and only the high-dose BHF group had prolonged swimming duration (P<0.05 or P<0.01). Compared to those in the low-dose BHF group, the bottoming time of the climbing pole were shorter in the moderate- and high-dose groups (P<0.05 or P<0.01), and the grip strength increased while the protein expression levels of TLR2, TLR4 and IL-17 decreased in the high-dose group (P<0.05 or P<0.01). The intestinal flora results showed significant differences between the blank group and the model group in the Dominance index, Pielou_e index, Shannon index, and Simpson index (P<0.05 or P<0.01). Compared to those of the model group, the Shannon index, Chao1 index, and Observed_otus index of the Madopar group, as well as the Chao1 index, Observed_otus index, Dominance index, Pielou_e index, Shannon index, and Simpson index of the high-dose BHF group all showed significantly statistical differences (P<0.05 or P<0.01). At the phylum level, the relative abundance categories of bacterial phyla with statistically significant differences in each group included Proteobacteria, Bacteroidetes, and Firmicutes (P<0.05 or P<0.01). At the genus level, the relative abundance categories of bacterial genera with statistically significant diffe-rences among each group included Muribaculaceae, Akkermansia, and Helicobacter pylori (P<0.05 or P<0.01). ConclusionThe possible mechanism of BHF in treating PD may be to reconstruct the disordered intestinal flora structure and improve the inflammatory response.