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Objective:To investigate the function, mechanism and therapeutic potential of macrophages in non-alcoholic steatohepatitis (NASH).Methods:Eight-week-old male foz/ foz (Alms mutant) mice were fed with a high fat diet (HFD) for 6, 8 and 10 weeks and 8-week-old male C57BL/6 mice were fed with a methionine and choline-deficient (MCD) diet for 7 d, 3 weeks and 4 weeks to establish NASH models. The mice of control group were fed with normal diet or MCD control diet. The expression of F4/80 mRNA level in the livers of mice of NASH model group and control group was detected by fluorescence quantitative polymerase chain reaction. Macrophages in the livers of mice of NASH group and control group were determined by immunofluorescence staining. After transgenic lysM-Cre/DTR mice were fed with MCD diet for 5 weeks, they were divided into transgenic experimental group (ablation of macrophages induced by diphtheria-toxin (DTox) injection) and transgenic control group (phosphate buffer saline injection). The levels of triglyceride and lipid peroxide in the livers of transgenic experimental group and transgenic control group were detected, and the inflammation of the livers of the mice was scored. The mechanism of macrophages regulating inflammation in NASH was investigated by cytokine profiliny analysis and Western blotting. The interaction between hepatocytes and macrophages were determined by co-culturing the conditional medium of hepatocytes AML-12 and macrophages RAW264.7. Macrophages of mice of control group and NASH model group were depleted by liposomal clodronate to confirm its value in NASH prevention. Independent sample t-test was used for statistical analysis. Results:F4/80 mRNA level in the livers of NASH model foz/ foz mice fed with HFD for 6 weeks, 8 weeks and 10 weeks was higher than that of control group (1.49±0.19, 1.70±0.15 and 1.93±0.04 vs.1.05±0.22), and the differences were statistically significant ( t=3.06, 4.92 and 7.92, all P<0.05). The expression of F4/80 mRNA level of the livers of NASH model mice fed with MCD for 7 d and 3 weeks was higher than that of control group (2.70±0.99 and 3.08±1.71 vs.1.00±0.83), and the differences were statistically significant ( t=3.43 and 3.54, both P<0.01). The results of immunofluorescence demonstrated that compared with that of control group, the number of F4/80 + inducible nitric oxide synthase (iNOS) + M1 macrophages were significantly increased, while F4/80 + CD206 + M2 macrophages were significantly decreased in the livers of NASH model mice fed with MCD for 4 weeks. After macrophages depletion, the inflammation score, the levels of triglyceride and lipid peroxide in the liver of transgenic experimental mice were all lower than those of transgenic control mice (0.69±0.32 vs. 1.95±0.74, (43.97±13.24) g/mg vs. (63.09±14.85) g/mg, (24.84±6.21) nmol/mg vs.(37.91±8.91) nmol/mg), and the differences were statistically significant ( t =3.14, 2.72 and 2.41, all P<0.05). The results of cytokine profiling analysis showed that macrophage depletion could lower the levels of interleukin (IL)-12 and macrophages inflammatory protein-1α (the difference between multiples: -3.98, -2.74, both P<0.05). CCAAT/enhancer binding protein β was defected in the nuclear of transgenetic experimental mice. In vitro study showed that RAW264.7 macrophages conditional medium could promote lipid accumulation in AML-12 hepatocytes, while conditional medium from MCD medium-treated AML-12 hepatocytes could promote RAW264.7 macrophages to M1 polarization. After treated with liposomal clodronate, the levels of triglyceride and lipid peroxidation in the liver of control mice were both lower than those of MCD-induced NASH model mice((45.33±14.59) g/mg vs. (63.10±16.02) g/mg, (2.11±0.48) nmol/mg vs. (2.73±0.17) nmol/mg), and the differences were statistically significant ( t=2.84 and 2.73, both P<0.05). The results of Western blotting indicated that after treating with liposomal clodronate, the relative content of phosphorylated protein kinase R-like endoplasmic reticulum kinase, inositol requiring enzyme-1α, protein disulfide isomerase, glucose regulatory protein 78, phosphorylated eukaryotic initiation factor 2α in the liver of NASH model mice were all lower than those of NASH model mice without liposomal clodronate treatment (1.84±0.36 vs. 3.05±0.83, 1.50±0.84 vs. 6.65±1.47, 0.87±0.12 vs. 2.28±0.52, 1.68±0.43 vs. 4.76±1.13, 1.42±0.19 vs. 2.75±0.79), and the differences were statistically significant( t=2.32, 5.28, 4.56, 4.41 and 2.85, all P<0.05). Conclusions:Macrophages are polarized into M1 phenotype in NASH. M1 macrophages contributed to NASH progression by interacting with hepatocyets to promote the secretion of inflammatory cytokines, up-regulation of lipogenic factors, oxidative stress and endoplasmic reticulum stress, resulting in the progression of NASH. Macrophages depletion by liposomal clodronate is a potential noval approach for NASH prevention.
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Objective:To observe any effect of intermittent theta burst stimulation (iTBS) of the cerebellum on swallowing dysfunction after cerebellar infarction, and to explore its mechanism.Methods:Sixty-two cerebellar stroke survivors with dysphagia were randomly divided into an observation group and a control group, each of 29. In addition to the routine swallowing rehabilitation training, the observation group was treated with iTBS, while the control group was given sham iTBS. The incubation and amplitude of the bilateral suprahyoid muscle motor evoked potential (MEP) were recorded before and after 4 weeks of treatment. The exponential approximate entropy (ApEn) of different brain regions was compared between the two groups during reflex and autonomous swallowing. Swallowing function was evaluated using the penetration-aspiration scale (PAS).Results:MEP incubation in the bilateral suprahyoid muscles had decreased significantly after 4 weeks of treatment in the observation group, and the MEP amplitude in the bilateral suprahyoid muscles of the two groups had increased significantly. The average improvement in the amplitude and incubation in the observation group was significantly greater than in the control group. The average ApEn at C3, C4, P3, P4, T5 and T6 had increased significantly in both groups during both reflex and spontaneous swallowing, with the improvement in the observation group significantly greater. Swallowing function had improved significantly in both groups, but the average PAS grade of the observation group was again significantly better.Conclusions:iTBS can improve the swallowing function of dysphagic cerebellar stroke survivors. This may be due to iTBS improving the excitability of the cerebral cortex and improving motor control of the swallowing muscles.
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Objective@#To explore the effect of Dahuang-Zhidan decoction on postoperative peripheral inflammatory factors and prognosis in patients with hypertensive cerebral hemorrhage.@*Methods@#A total of 103 patients with hypertensive cerebral hemorrhage in our hospital from May 2015 to January 2017 were divided into two groups according to the randomly number tables method. The control group (n=51) were treated by conventional treatment of western medicine, and the observation group (n=52) were treated with Dahuang-Zhidan decoction based on the treatment of the control group.The inflammatory factors, including interleukin-6 (IL-6), tumor necrosis factor (TNF-α), highly sensitive C-reactive protein (hs-CRP), interleukin-1β (IL-1β) in hematoma lesions after treatment for 1st, 7th day between two groups were measured. The comparisons of awake rate after treatment for the 7th, 14th, 30th day between two groups was conducted. The National Institute of Health Stroke Scale (NIHSS) scores after treatment for 30th, 90th day between two groups was launched. The Glasgow Prognosis Score (GOS) between two groups was compared at the 90th day.@*Results@#After treatment for 7 d,the postoperative peripheral inflammatory factors IL-6 (90.55 ± 18.89 pg/ml vs. 122.71 ± 19.62 pg/ml, t=-6.892), TNF-α (172.26 ± 29.85 pg/ml vs. 229.02 ± 30.34 pg/ml, t=-10.332), hs-CRP (137.55 ± 24.60 pg/ml vs. 162.14 ± 23.98 pg/ml, t=-8.615), IL-1β (95.57 ± 18.92 pg/ml vs. 110.65 ± 17.38 pg/ml, t=-3.594) in the observation group was significantly lower than those in the control group (P<0.05). After treatment for 7, 14, 30 d, the awake rate was 44.23% (23/52), 69.23% (36/52), 90.38% (47/52) in the observation group, which was significantly higher than those in the control group (χ2=3.977, 4.355, 4.499, P<0.05). After treatment for 30, 90 d, the NIHSS scores (12.24 ± 6.38 vs. 16.27 ± 7.69, t=-2.906), (5.39 ± 6.24 vs. 9.94 ± 8.11 , t=-3.198) in the observation group were significantly lower than those in the control group (P<0.05). After treatment for 90 d, the distribution of GOS scores in the observation group was significantly different from the control group (Z=-2.400, P=0.016).@*Conclusions@#The Dahuang-Zhidan decoction in the treatment of patients with hypertensive cerebral hemorrhage significantly inhibit postoperative the inflammatory response around hematoma lesions, and improve the neurological function, the awake rate and the qualify of life.
