OBJECTIVES: To explore the mechanisms of Qingre Lidan Jiedu Recipe (QLJR) for improving cognitive dysfunction in rats with high copper load. METHODS: Seventy-five male SD rats were randomized into normal control group, model group, QLJR group, penicillamine (PCA) group, and QLJR+ PCA group. Except for those in the control group, all the rats were fed a high-copper diet for 12 weeks. The effects of the treatments on cognitive function of the rats were assessed using the Barnes maze and passive avoidance tests. Hippocampal expressions of NIX, FUNDC1 and LC3 of the rats were detected using Western blotting and immunofluorescence staining, and changes in mitochondrial morphology were observed with transmission electron microscopy. RESULTS: Behavioral tests showed prolonged target hole latency, shortened latency to enter the dark chamber, and increased error counts of the rats in the model group, which were significantly improved in QLJR+PCA group; the error counts were significantly lower in QLJR+PCA group than in either QLJR or PCA group. Among all the groups, the hippocampal expressions of NIX and FUNDC1 were the lowest and LC3 I/II expression the highest in the model group; NIX and FUNDC1 expressions were significantly higher and LC3 I expression was lower in QLJR+PCA group than in QLJR group and PCA group. Immunofluorescence staining revealed weakened NIX and FUNDC1 expressions and enhanced LC3 expression in the hippocampus of the rats in the model group as compared with those in the normal control and QLJR+PCA groups, but their expressions did not differ significantly between QLJR and PCA groups. The rats in the model group showed obvious structural disarray of the mitochondria, which were improved in all the treatment groups. CONCLUSIONS QLJR improves cognitive dysfunction in rats with high copper load possibly by regulating mitophagy.
Animals ; Male ; Rats, Sprague-Dawley ; Rats ; Drugs, Chinese Herbal/therapeutic use* ; Copper/toxicity* ; Mitophagy/drug effects* ; Hippocampus/drug effects* ; Cognition Disorders/drug therapy* ; Cognitive Dysfunction/chemically induced*

Objective To investigate the repairing effect and mechanism of Chinese patent medicine Weiyangning pill on gastric mucosal injury in rats induced by anhydrous ethanol,and to establish a high-performance liquid chromatography(HPLC)method to determine the five main components of Weiyangning pill.Methods The five components of paeoniflorin,psoralen,atractylenolide Ⅲ,liquiritin and hesperidin in Weiyangning pill were detected by HPLC.SD male rats were randomly divided into normal control group,model control group,Weinaian group,and large and small dose group of Weiyangning pill.All rats were fasted for 24 hours without water fasting.The normal control group and the model control group were given purified water by gavage.While Weinaian group was given Weinaian(3 g·kg-1),the test group were given intragastric perfusion of Weiyangning(3,1.5 g·kg-1)respectively.After 2 hours,all the rats,except the normal control group,were intragastrically administered with anhydrous ethanol(5 mL·kg-1)to establish the model of gastric mucosal injury.An hour later,the experimental materials were collected,and the gross score of gastric mucosal injury was observed and calculated.The gastric mucosal slices were stained by hematoxylin-eosin(HE)to calculate the pathological scores.Immunohistochemistry was employed to detect the expression of gastric mucosa-related proteins.Results The high-performance liquid chromatogram of Weiyangning pill was obtained,and the absorption peaks with the same retention time as the five standard substances(paeoniflorin,psoralen,atractylenolide Ⅲ,liquiritin and hesperidin)were observed.The general score and pathological score of gastric mucosal injury in Weiyangning groups(3,1.5 g·kg-1)were lower than those of the model control group(P<0.05 or P<0.01).Weiyangning pill(3,1.5 g·kg-1)ameliorated the decrease expression of tight junction protein(Claudin-7),adhesion junction proteins(E-cadherin and β-catenin),mucins(MUC1 and MUC5AC),and the gastric transcription factor SOX2 in the gastric mucosa of the rats modeled in anhydrous ethanol(P<0.05 or P<0.01 compared with the model control group).Conclusion The repairing effect of Weiyangning pill on gastric mucosal injury induced by anhydrous ethanol in rats is related to the increase of the expression of tight junction protein,adhesion junction protein,mucin and gastric transcription factor.

Objective:To investigate the form of programmed cell death (PCD) induced by severe fever with thrombocytopenia syndrome virus (SFTSV) infection in Vero cells and further explore the existence of pyroptosis, so as to provide new ideas for studying the pathogenic mechanism of SFTSV.Methods:Vero cells were infected with SFTSV at different multiplicity of infection (MOI), cytopathic effect (CPE) was observed daily, cell viability was detected by CCK-8 method, and cell membrane damage was detected by LDH release test to determine the optimal amount of virus infection and cell death time. Vero cells were pretreated with different PCD inhibitors and infected with SFTSV. CCK-8 kit was used to detect the cell viability and determine the death form of PCD caused by SFTSV. The expression of pyroptosis related proteins was detected by Western blotting to further explore the existence of pyroptosis.Results:At 48 h and 72 h after SFTSV infected Vero cells with MOI=10, the optimal infection amount and time of subsequent experiments were observed. At this time, the CPE of cells was obvious, the cell viability decreased to 51% and 41% of the control group ( P<0.001, P<0.001), and the LDH release amount reached 24% and 37% of the maximum enzyme activity release wells, were 3.8, 3.4 times of LDH release in the control group ( P<0.001, P<0.001). The inhibition of SFTSV-induced cell death by different PCD inhibitors showed that pan-caspase inhibitor and receptor-interacting serine/threonine-protein kinase 3 (RIP3) inhibitor had inhibitory effects at 48 h and 72 h. The cell viability was 2.1 and 1.6 times of the viral control group at 48 h, 2.3 and 1.7 times of the viral control group at 72 h, and the effect of pan-caspase inhibitor was significantly higher than that of the other inhibitor groups. Caspase-1 and caspase-3 inhibitors only had inhibitory effect at 48 h, and the cell viability was 1.5 and 1.3 times of the viral control group. After SFTSV infection of Vero cells, the expressions of caspase-1 and IL-1β increased gradually with the prolongation of time, and reached 3.4 and 9.5 times of the control group at 48 h, respectively ( P<0.001, P<0.001). Conclusions:SFTSV infection of Vero cells can lead to various forms of PCD, including apoptosis, pyroptosis and programmed necrosis, and pyroptosis related protein activation can be detected in the process of PCD, which further explored the existence of pyroptosis.

Objective:To study the inactivation effects of microwave on human adenovirus 2 (HAdV-2) in simulated infectied wastes, and to explore its molecular mechanism.Methods:25 μl of HAdV-2 virus suspension was dripped with medical disposable gloves, masks and cotton swabs to simulate infectied wastes, and irradiated under different microwave conditions: gloves and masks were irradiated for 30 s, 60 s, and 90 s at 300 W, 500 W, and 700 W, respectively. Cotton swabs irradiate 60 s, 90 s, 120 s at 500 W and 700 W respectively. Temperature changes were recorded, and the inactivation logarithmic values were calculated by the 50% endpoint method to evaluate the microwave inactivation effects, and the proliferation ability of the virus was detected by qPCR. The damage of Penton, Fiber, Hexon and E2 B genes was detected by PCR. The virus was treated with the highest temperature of 76 ℃ during microwave irradiation to study whether there was non-thermal effect during microwave disinfection. Results:After microwave irradiation of infectied waste, the temperature of masks and gloves carriers rises rapidly, with the highest temperature of 76 ℃. The temperature of the cotton swab carriers rose slowly, and the highest temperature is 65 ℃. The inactivation effect of microwave on HAdV-2 was positively correlated with microwave power and irradiation time. In the mask and glove group, microwave power of 700 W irradiated for 60 seconds, and the inactivation logarithm value could reach 3.0, In the cotton swab group, microwave power of 700 W irradiated for 120 seconds, and the inactivation logarithm value was still less than 3.0. This indicated that there were differences in the conditions for microwave inactivation of the virus in different carriers. The qPCR result showed that microwave irradiation could weak the proliferation ability of the virus. Microwave irradiation had no effect on the virus's Penton and Fiber genes, but caused some damage to the Hexon and E2 B genes. The inactivation effect of individual heat treatment on HAdV-2 was weaker than that of microwave irradiation, and there was no damage to the Hexon, Penton, Fiber, and E2 B genes. This indicated the presence of non thermal effects during the microwave inactivation process. Conclusions:Microwave irradiation can inactivate HAdV-2 in simulated infectied wastes through thermal and non-thermal effects, and its damage to viral DNA is one of the mechanisms of virus inactivation.