Koumine regulates macrophage M1/M2 polarization via TSPO, alleviating sepsis-associated liver injury in mice.

BACKGROUND: Translocator protein (TSPO) is an 18-kDa transmembrane protein found primarily in the mitochondrial outer membrane, and it is implicated in inflammatory responses, such as cytokine release. Koumine (KM) is an indole alkaloid extracted from Gelsemium elegans Benth. It has been reported to be a high-affinity ligand of TSPO and to exert anti-inflammatory and immunomodulatory effects in our recent studies. However, the protective effect of KM on sepsis-associated liver injury (SALI) and its mechanisms are unknown. PURPOSE: To explore the role of TSPO in SALI and then further explore the protective effect and mechanism of KM on SALI. METHODS: The effect of KM on the survival rate of septic mice was confirmed in mouse models of caecal ligation and puncture (CLP)-induced and lipopolysaccharide (LPS)-induced sepsis. The protective effect of KM on CLP-induced SALI was comprehensively evaluated by observing the morphology of the mouse liver and measuring liver injury markers. The serum cytokine content was detected in mice by flow cytometry. Macrophage polarization in the liver was examined using western blotting. TSPO knockout mice were used to explore the role of TSPO in sepsis liver injury and verify the protective effect of KM on sepsis liver injury through TSPO. RESULTS: KM significantly improved the survival rate of both LPS- and CLP-induced sepsis in mice. KM has a significant liver protective effect on CLP-induced sepsis in mice. KM treatment ameliorated liver ischaemia, improved liver pathological injuries, and decreased the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and proinflammatory cytokines in serum. Western blotting results showed that KM inhibited M1 polarization of macrophages and promoted M2 polarization. In TSPO knockout mice, we found that TSPO knockout can improve the survival rate of septic mice, ameliorate liver ischaemia, improve liver pathological injuries, and decrease the levels of ALT, AST, and LDH. In addition, TSPO knockout inhibits the M1 polarization of macrophages in the liver of septic mice and promotes M2 polarization and the serum levels of proinflammatory cytokines. Interestingly, in TSPO knockout septic mice, these protective effects of KM were no longer effective. CONCLUSIONS: We report for the first time that TSPO plays a critical role in sepsis-associated liver injury by regulating the polarization of liver macrophages and reducing the inflammatory response. KM, a TSPO ligand, is a potentially desirable candidate for the treatment of SALI that may regulate macrophage M1/M2 polarization through TSPO in the liver.

[Effects of day and night temperature difference on growth, development, yield and fruit quality of tomatoes].

Abstract: The effects of day and night temperature difference (DIF) on tomato's growth were studied in three precisely controlled units in phytotron. Set DIF as 6 °C (25/19 C), 8 °C (26/18 °C), 10 °C (27/17 °C) respectively, with the same diurnal mean temperature as 22 °C. The results showed that, different tomato varieties needed different suitable DIF at different growth stages. Before flouring, compared with DIF 6 °C , DIF 8 °C could significantly improve the growth and development of the wild currant tomato LA1781, increasing the plant height by 23.1%, fastening leaf development by 1-2 leaves, advancing flowers by 7 d. DIF 10 °C had similar effects with DIF 8 °C on LA1781. As to the cultured ordinary tomatoes LA2397 and LA0490, DIF 6 °C made the seedlings grow well, DIF 8 °C had no significant improved effects on seedlings, DIF 10 °C depressed the seedling's growth and flouring, decreasing the plant height by 12.0%-18.3%, lowering the leaf development by 2-3 leaves, delaying flouring by 2-4 d. But DIF 10 °C increased the dry aboveground mass of these three varieties by 25.2%-44.2%. After flouring, compared with DIF 6 °C, DIF 10 °C could significantly improve the yield and fruit quality of LA1781, increasing fruit number by 34.7%, yield per plant by 92.1%, single fruit mass by 40.0%, soluble sugar content by 16.3%, lycopene content by 95.6%. Compared with DIF 6 °C, LA2397 and LA0490 had higher yields and better fruit quality under DIF 8 °C, and lycopene content increased more than twice as that under DIF 6 °C. Under DIF 10 °C, yields of LA2397 and LA0490 slightly decreased (5.0%), soluble sugar contents of fruit decreased, but fruit size and lycopene content increased. The results showed that, DIF should not be very great in the seedling period of tomatoes, and a moderate DIF in flower and fruit periods could improve the yield and fruit quality, but a too high DIF would result in poor growth and yield reduction.

Relationship between nm23H1 genetic instability and clinical pathological characteristics in Chinese digestive system cancer patients.

AIM: To study the relationship between nm23H1 gene genetic instability and its clinical pathological characteristics in Chinese digestive system cancer patients. METHODS: Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze the microsatellite instability (MSI) and loss of heterozygosity (LOH). Immunohistochemistry was employed to detect the expression of nm23H1. RESULTS: The MSI was higher in TNM stage I + II than in stage III + IV of gastric, colonic and gallbladder carcinomas. The LOH was higher in TNM stage III + IV than in stage I + II of gastric, colonic and hepatocellular carcinomas. Lymphatic metastasis was also observed. The expression of nm23H1 protein was lower in TNM stage III + IV than in stage I + II of these tumors and in patients with lymphatic metastasis.The nm23H1 protein expression was higher in the LOH negative group than in the LOH positive group. CONCLUSION: MSI and LOH may independently control the biological behaviors of digestive system cancers. MSI could serve as an early biological marker of digestive system cancers. Enhanced expression of nm23H1 protein could efficiently inhibit cancer metastasis and improve its prognosis. LOH mostly appears in late digestive system cancer.

[Protective effect of Danzhi-xiaoyao San on rat brain energy or material metabolism (correction of matebolism) dealt with D-galactose].

OBJECTIVE: To research the mechanism of Danzhi-xiaoyao San (DZXYS) for treating Alzheimer's disease model of rats dealt with D-galactose. METHOD: An Alzheimer's disease-like model of rats has been set up with sc. D-galactose 150.0 mg kg-1 D-1 x 49 d. Comparing with Acricept in 0.54 mg kg-1 D-1 dosage as a positive control drug, DZXYS in 12.636 g kg-1 D-1 x 49 d dosage has orally been administrated orally to treat the injury in the Alzheimer's disease-like model of rats. The energy charge in the cerebral tissues had been detested with waters liquid chromatography; the protein content and DNA content in the cerebral tissues had been detested with ultraviolet assay, the relative content of aldose reductase-mRNA is detested with RT-PCR. The difference was analyzed between the control rats without D-galactose, the model rats dealt with D-galactose, the model rats treated with Aricept and the model rats treated with DZXYS, it is significant as P<0.05. RESULT: 1) DZXYS can not affect the energy charge in their cerebral tissues. 2) DZXYS can increase the protein content from 0.3139 +/- 0.019468 to 0.3213 +/- 0.015528 (ni=10, P>0.05) in their cerebral tissues. 3) DZXYS can increase the total DNA content from 1.093 +/- 0.267 to 1.488 +/- 0.341 (ni=10, P<0.01) in their cerebral tissues. 4) DZXYS can increase the content of AR-mRNA in their cerebral tissue from 0.732 +/- 0.159 to 1.418 +/- 0.277 (ni=5, P<0.01). CONCLUSION: It suggests that DZXYS could be effective in human Alzheimer's disease for its stabling gene expression, maintaining protein characteristics, recovering signal transduction in the Alzheimer's disease-like model rats dealt with D-galactose.

[Study on cardiac gene expression microarray and recipe for activating blood circulation and supplementing qi on heart of rats with post-infarction heart failure].

OBJECTIVE: To observe the effect of recipe for activating blood circulation and supplementing Qi (RAS) on cardiac functional structure in rats with post-infarction heart failure (PIHF). METHODS: Rat model of PIHF was established by left coronary artery ligation. Left ventricular samples of model rats from infarcted or peri-infarcted area were obtained at PIHF formation stage and stable stage (10 days and 8 weeks respectively after operation), the total RNA extracted and detected using 6 pieces of rat's 40s gene microarray (4096 genes/microarray), the data were analyzed using software as Genespring, Treeview, Clustering and SOM. Besides, RAS was used to treat the model rats beginning from 4 weeks after modeling and lasted for 4 weeks, changes of heart function and cardiac coefficient before and after treatment were observed by impedance method with Captopril as positive control. RESULTS: (1) Genespring analysis showed thousands of genes differential expression (upper or down regulated), including 13 kinds of gene involving energy metabolism, myocardial cytoskeleton, fibrosis, etc. which, in the infarcted area at heart formation stage were 1086 genes and at the stable stage, 724 genes, while in the peri-infarcted area, formation stage 196 genes and stable stage 97 genes. (2) After RAS or Captopril treatment, the heart function improved significantly, with the stroke volume, cardiac output and cardiac index increased significantly (P < 0.01). RAS could also improve the cardiac coefficient of model rats, as compared with that in untreated model, P < 0.01, compared with that in the sham-operated rats, P < 0.05. CONCLUSION: PIHF is a kind of overload heart disease with multiple genes abnormality. RAS could improve the heart function and histologic indexes, so as to treat the heart failure.

[Different effects of illumination with xenon and sulfur lamp on growth and development of cotton plants].

Experiments were carried out with cotton (Gossgpium hirsutum cv. Xuzhou 142) plants to study the effects of illumination with xenon and sulfur lamp on development of cotton plants. The results showed that, compared with xenon lamp, illumination with sulfur lamp inhibited excessive elongation of hypocotyl via promotion of longitudinal elongation of epidermis and cortex cells, increased the numbers of branches, buds and bolls significantly. It suggested that illumination with sulfur lamp rendered cotton photomorphogenesis more favorable to yield formation.

[Effect of different drug dosage to activate blood circulation and to nourish qi on cardiac function and structure of congestive heart failure rats after acute myocardial infarction].

OBJECTIVE: To observe the effect of different drug dosage to activate blood circulation and to nourish Qi on cardiac function and structure of heart failure rats after acute myocardial infarction. METHOD: The congestive heart failure post cardiac infarction rats models subjected to left coronary occlusion were made and given different dosage of drug of Huoxue Yiqi and captopril (positive control) from 4 weeks to 8 weeks after operation. The cardiac function before and after using drugs were observed by impedance methods. RESULT: Treated by different dosage of drug of Huoxue Yiqi and captopil, cardiac function of heart failure rats after acute myocardial infarction improved, SV, CO, CI(P < 0.01); at the same time, prescription of Huoxue Yiqi and Qixue could improve the model rats' heart coefficient, VS SHAM(P > 0.05), but prescription of Yiqi Huoxue and captopril could not improve it. CONCLUSION: Drug to activate blood circulation and to increase qi can treat heart failure rats after acute myocardial infarction, through improving their function and structure. At the same time, more dosage of drug to activate blood circulation and to increase qi(Huoxue/Yiqi drug) may improve the model rats' heart coefficient.