Apolipoprotein A-1 protected hepatic ischaemia-reperfusion injury through suppressing macrophage pyroptosis via TLR4-NF-κB pathway.

BACKGROUND AND AIMS: Apolipoprotein A-1 (ApoA-1), the major apolipoprotein of high-density lipoprotein, plays anti-atherogenic role in cardiovascular diseases and exerts anti-inflammation effect in various inflammatory and infectious diseases. However, the role and mechanism of ApoA-1 in hepatic ischaemia-reperfusion (I/R) injury is unknown. METHODS: In this study, we measured ApoA-1 expression in human liver grafts after transplantation. Mice partial hepatic I/R injury model was made in ApoA-1 knockout mice, ApoA-1 mimetic peptide D-4F treatment mice and corresponding control mice to examine the effect of ApoA-1 on liver damage, inflammation response and cell death. Primary hepatocytes and macrophages were isolated for in vitro study. RESULTS: The results showed that ApoA-1 expression was down-regulated in human liver grafts after transplantation and mice livers subjected to hepatic I/R injury. ApoA-1 deficiency aggravated liver damage and inflammation response induced by hepatic I/R injury. Interestingly, we found that ApoA-1 deficiency increased pyroptosis instead of apoptosis during acute phase of hepatic I/R injury, which mainly occurred in macrophages rather than hepatocytes. The inhibition of pyroptosis compensated for the adverse impact of ApoA-1 deficiency. Furthermore, the up-regulated pyroptosis process was testified to be mediated by ApoA-1 through TLR4-NF-κB pathway and TLR4 inhibition significantly improved hepatic I/R injury. In addition, we confirmed that D-4F ameliorated hepatic I/R injury. CONCLUSIONS: Our study has identified the protective role of ApoA-1 in hepatic I/R injury through inhibiting pyroptosis in macrophages via TLR4-NF-κB pathway. The effect of ApoA-1 may provide a novel therapeutic approach for hepatic I/R injury.

Apolipoprotein A-1 Accelerated Liver Regeneration Through Regulating Autophagy Via AMPK-ULK1 Pathway.

BACKGROUND & AIMS: Apolipoprotein A-1 (ApoA-1), the main apolipoprotein of high-density lipoprotein, has been well studied in the area of lipid metabolism and cardiovascular diseases. In this project, we clarify the function and mechanism of ApoA-1 in liver regeneration. METHODS: Seventy percent of partial hepatectomy was applied in male ApoA-1 knockout mice and wild-type mice to investigate the effects of ApoA-1 on liver regeneration. D-4F (ApoA-1 mimetic peptide), autophagy activator, and AMPK activator were used to explore the mechanism of ApoA-1 on liver regeneration. RESULTS: We demonstrated that ApoA-1 levels were highly expressed during the early stage of liver regeneration. ApoA-1 deficiency greatly impaired liver regeneration after hepatectomy. Meanwhile, we found that ApoA-1 deficiency inhibited autophagy during liver regeneration. The activation of autophagy protected against ApoA-1 deficiency in inhibiting liver regeneration. Furthermore, ApoA-1 deficiency impaired autophagy through AMPK-ULK1 pathway, and AMPK activation significantly improved liver regeneration. The administration of D-4F could accelerated liver regeneration after hepatectomy. CONCLUSIONS: These findings suggested that ApoA-1 played an essential role in liver regeneration through promoting autophagy in hepatocytes via AMPK-ULK1 pathway. Our findings enrich the understanding of the underlying mechanism of liver regeneration and provide a potential therapeutic strategy for liver injury.

[Transformation and Distribution of Soil Organic Carbon and the Microbial Characteristics in Response to Different Exogenous Carbon Input Levels in Paddy Soil].

The turnover of soil organic carbon (SOC) and the activity of soil microbes can be influenced by exogenous carbon. However, microbial response characteristics of the transformation and distribution of available organic carbon under different levels remain unclear in paddy soils. 13C-labeled glucose was used as a typical available exogenous carbon to simulate indoor culture experiments added at different levels of soil microbial biomass carbon (MBC) (0×MBC, 0.5×MBC, 1×MBC, 3×MBC, and 5×MBC) to reveal the process of C-transformation and distribution. The characteristics of microbial response in the process of exogenous carbon turnover was also monitored. The 96-well microplate fluorescence analysis was adopted to determine the activities of cellobiose hydrolase (CBH) and ß-glucosidase (ß-Glu). The results showed that, in 2 d of incubation, the ratio of labeled glucose carbon to dissolved organic carbon (13C-DOC/DOC) or to SOC (13C-SOC/SOC) was positively correlated with the amount of glucose added. The incorporation of glucose C (13C) into MBC reached the highest value (18.96 mg·kg-1) at 3×MBC treatment but decreased thereafter. The 13C allocation rate was mainly positively correlated with MBC, Olsen-P, and DOC. At 60 d, 13C-DOC, 13C-MBC, and 13C-SOC decreased significantly to less than 0.02 mg·kg-1, 2 mg·kg-1, and 10 mg·kg-1 in soil, and it was positively correlated with the amount of glucose added. Compared with CK, CBH enzyme activity increased significantly after the addition of glucose, and for the 3×MBC treatment it was increased by 22.6 times, which was significantly higher than those of other treatments (P<0.05). However, ß-Glu enzyme activity increased only in the 3×MBC and 5×MBC treatments, wherein it decreased with increasing amounts of added glucose. NH4+-N, pH, ß-Glu, and CBH were the primary factors affecting the distribution rate of 13C. In conclusion, the conversion of exogenous carbon to SOC increased with increased amounts of added organic carbon. This changed the activity of soil enzymes; however, microbial utilization of exogenous carbon may have a saturation threshold. Within the saturation threshold, the conversion rate of organic matter was directly proportional to the amount of added organic matter. When the saturation threshold was exceeded, the conversion rate of organic matter decreased. Therefore, the appropriate addition of exogenous carbon is beneficial, as it can increase SOC in rice fields and improve the quality of the crop growth environment.

[Characterization of Soil Organic Carbon Mineralization Under Different Gradient Carbon Loading in Paddy Soil].

Available carbon is the most active part of the soil carbon pool. It is also the main carbon source of soil microbes and plays an important role in the processes of soil organic carbon mineralization and accumulation. However, the mechanisms are still not clear how soil organic carbon mineralization and its priming effect (PE) are affected by different input levels of readily available carbon, based on the growth requirements of microbes in paddy soil. In this study, an incubation experiment was conducted by adding different levels (0.5, 1, 3, and 5 times of MBC) of exogenous source organic carbon (13C-glucose) to the soil. The mineralization dynamics of labile organic carbon and its priming effect was investigated. The mineralization rate of glucose-C increased significantly with the increasing carbon loading level. The distribution of glucose-C into rapid and slow C pools was also exponentially correlated with the carbon loading (R2=0.99, P<0.05 and R2=0.99, P<0.05, respectively). Negative PE was observed at high carbon loading (3×MBC and 5×MBC); while positive PE was induced by low carbon loading (0.5×MBC and 1×MBC). The cumulative PE was 160.0 mg·kg-1 and 325.1 mg·kg-1, respectively, at the end of the incubation. Redundancy analysis showed that the main factors affecting the cumulative PE were MBC, MBN, and DOC at the initial glucose mineralization stage, while ß-glucosidase, chitinase, and ammonium nitrogen were the main factors at later stages. Therefore, the readily available carbon loading has an important effect on the organic carbon mineralization and PE in paddy soil. Higher carbon loading was good for the accumulation of organic carbon sequestration in paddy soil. This study is of great scientific significance for revealing the activity of organic carbon in paddy fields and for its contribution to the development of sustainable agriculture.

Genetic diversity of Haemonchus contortus isolated from sympatric wild blue sheep (Pseudois nayaur) and sheep in Helan Mountains, China.

BACKGROUND: Haemonchus contortus is known among parasitic nematodes as one of the major veterinary pathogens of small ruminants and results in great economic losses worldwide. Human activities, such as the sympatric grazing of wild with domestic animals, may place susceptible wildlife hosts at risk of increased prevalence and infection intensity with this common small ruminant parasite. Studies on phylogenetic factors of H. contortus should assist in defining the amount of the impact of anthropogenic factors on the extent of sharing of agents such as this nematode between domestic animals and wildlife. METHODS: H. contortus specimens (n = 57) were isolated from wild blue sheep (Pseudois nayaur) inhabiting Helan Mountains (HM), China and additional H. contortus specimens (n = 20) were isolated from domestic sheep that were grazed near the natural habitat of the blue sheep. Complete ITS2 (second internal transcribed spacer) sequences and partial sequences of the nad4 (nicotinamide dehydrogenase subunit 4 gene) gene were amplified to determine the sequence variations and population genetic diversities between these two populations. Also, 142 nad4 haplotype sequences of H. contortus from seven other geographical regions of China were retrieved from database to further examine the H. contortus population structure. RESULTS: Sequence analysis revealed 10 genotypes (ITS2) and 73 haplotypes (nad4) among the 77 specimens, with nucleotide diversities of 0.007 and 0.021, respectively, similar to previous studies in other countries, such as Pakistan, Malaysia and Yemen. Phylogenetic analyses (BI, MP, NJ) of nad4 sequences showed that there were no noticeable boundaries among H. contortus populations from different geographical origin and population genetic analyses revealed that most of the variation (94.21%) occurred within H. contortus populations. All phylogenetic analyses indicated that there was little genetic differentiation but a high degree of gene flow among the H. contortus populations among wild blue sheep and domestic ruminants in China. CONCLUSIONS: The current work is the first genetic characterization of H. contortus isolated from wild blue sheep in the Helan Mountains region. The results revealed a low genetic differentiation and high degree of gene flow between the H. contortus populations from sympatric wild blue sheep and domestic sheep, indicating regular cross-infection between the sympatrically reared ruminants.