Abstract Hepatic ischemia/reperfusion injury (IRI) is tissue damage resulting from return of the blood supply to the tissue after a period of ischemia or lack of oxygen. Much of the morbidity associated with liver transplantation and major hepatic resections is, in part, due to IRI. Both innate immunity and autophagy play important roles in hepatic IRI. With regard to innate immunity, one factor that plays a key role is NOD1, an intracellular pattern recognition receptor. NOD1 has recently been shown to be associated with autophagy, but the mechanisms involved with this process remain obscure. This relationship between NOD1 and autophagy prompted us to examine the role and potential mechanisms of NOD1 in regulating autophagy as related to hepatic IRI. We found that NOD1 was upregulated during hepatic IRI and was associated with an activation of the autophagic signaling pathway. Moreover, levels of Atg5, a critical protein associated with autophagy, were decreased when NOD1 was inhibited by NOD1 small interfering RNA. We conclude that NOD1 appears to exert a pivotal role in hepatic IRI by activating autophagy to aggravate hepatic IRI, and Atg5 was required for this process. The identification of this novel pathway, that links expression levels of NOD1 with Atg5‐mediated autophagy, may provide new insights for the generation of novel protective therapies directed against hepatic IRI.
Content RFamide‐related peptide‐3 (RFRP‐3), the mammalian ortholog of gonadotropin‐inhibiting hormone, has been implicated as a mediator between reproduction and energy balance. This study aimed to investigate the physiological effects of RFRP‐3 on the process of ovarian development in food‐restricted pre‐pubertal ewes. The results showed that food restriction significantly inhibited the ovarian development and follicular growth. The data of qPCR in the hypothalamic–pituitary–ovarian (HPO) axis showed that food restriction not only upregulated RFRP‐3 mRNA expression but also downregulated the mRNA expression of gonadotropin‐releasing‐hormone receptor, follicle‐stimulating hormone receptor and luteinizing hormone receptor (LHR). Immunohistochemistry of RFRP‐3 in the ovaries suggested that RFRP‐3 may regulate the follicular development. These results suggested that the changes of RFRP‐3 in response to food restriction might influence the HPO axis and inhibit ovarian development.
Recent study found psychosocial factors play some important roles in carcinogenesis and development of malignant tumors, but its mechanisms remain unclear. This study was to investigate the impact of chronic restraint stress on splenocyte immunity and growth of mouse forestomach carcinoma (MFC) xenografts in Kunming mice, and provide evidences for exploring the mechanisms of psychosocial factors function on malignant tumors.A total of 60 Kunming mice were randomized into normal control group, restraint stress group, tumor-bearing group and tumor plus restraint stress group; each group contained 15 mice. Chronic restraint stress models were established in restraint stress group and tumor plus restraint stress group; MFC xenograft models were established in tumor-bearing group and tumor plus restraint stress group 4 weeks later. Mice were killed 10 days after inoculation of MFC cells. The weight of MFC xenografts were measured. The proliferation and cytotoxicity of splenocytes were detected by MTT assay. The level of interleukin-2 (IL-2) in splenocyte culture supernants was detected by enzyme-linked immunoabsorbent assay (ELISA).The weight of MFC xenografts was (1.39+/-0.39) g in tumor-bearing group and (2.10+/-0.52) g in tumor plus restraint stress group; MFC xenografts grew faster in tumor plus restraint stress group than in tumor-bearing group (P<0.01), with a tumor growth rate of 51.08%. In normal control group, restraint stress group, tumor-bearing group, and tumor plus restraint stress group, the stimulus indexes (SI) of T lymphocytes were 1.77+/-0.22, 1.70+/-0.17, 1.69+/-0.18, and 1.22+/-0.15, respectively; the SI of B lymphocytes were 1.73+/-0.14, 1.65+/-0.17, 1.64+/-0.21, and 1.33+/-0.11, respectively; the inhibition rate of MFC cell proliferation were (23.01+/-4.76)%, (19.47+/-3.70)%, (16.81+/-3.68)%, and (7.14+/-5.00)%, respectively, when the effector/target ratio was 5:1 and (33.03+/-3.91)%, (28.34+/-4.58)%, (24.94+/-2.97)%, and (13.49+/-7.94)%, respectively, when the effector/target ratio was 10:1; the levels of IL-2 in splenocyte culture supernatants were (260.03+/-14.96) pg/mL, (239.78+/-10.93) pg/mL, (238.11+/-13.50) pg/mL, and (186.34+/-10.42) pg/mL, respectively. Both chronic restraint stress and MFC xenografts impaired the proliferation, cytotoxicity, and IL-2 secretion of splenocytes; the two factors showed interactive effect (P<0.01), but the effect of chronic restraint stress was much more obvious.Chronic restraint stress may impair the immune function and promote the growth of MFC xenografts in mice.
To understand the underlying mechanism(s) for the effect of exercise at different intensities on T cell and DNA vaccination responses, we treated mice in a training protocol with regular moderate‐intensity exercise (MIE) or prolonged, exhaustive high‐intensity exercise (HIE). After 6 weeks of training, splenocytes were isolated to evaluate cytokine expression and T‐regulatory (Treg) cell proportion by RT‐PCR and FACS, respectively. Another set of mice that completed the same training protocol were used to determine DNA vaccination responses. These mice were immunized three times with HBV DNA vaccine at 2‐week intervals and euthanized on day 14 after the last immunization. Serum and splenocytes were isolated to determine humoral and cell‐mediated immunity (CMI). Results showed that HIE increased anti‐inflammatory cytokine expression and CD4 + CD25 + Treg cell proportion. Further, HIE decreased IFN‐γ expression, T‐lymphocyte proliferation, and antigen‐specic cytotoxic response in HBV DNA vaccine‐immunized mice. MIE did not change anti‐inflammatory cytokine expression or CD4 + CD25 + Treg cell proportion but increased pro‐inflammatory cytokine expression and augmented antigen‐specific CMI. Thus, MIE lower the risk of cancer and infectious illness through enhancing the pro‐inflammatory responses. By contrast, HIE might increase the risk of common infections, such as upper respiratory tract infection, due to an up‐regulation of CD4 + CD25 + Treg cells and anti‐inflammatory responses.
Objective
To investigate the effect of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) on pyroptosis in hepatic ischemia-reperfusion injury.
Methods
An animal model of ischemia-reperfusion injury was established. Thirty healthy, male, and clean C57 BL mice were randomly divided into sham operation group (sham group) and ischemia-reperfusion group (IR, including 2 h, 6 h, 12 h, 24 h subgroups), 6 per group. Serum ALT and AST levels in each group were measured by blood biochemistry. HE staining and TUNEL were used to observe the pathological changes of liver and hepatocyte apoptosis. Immunohistochemistry was used to detect the expression and distribution of NOD1 in each group. Western blotting was used to detect NOD1, AIM2, pro-Caspase-1 and active-Caspase-1 expression. NOD1 siRNA and empty control siRNA were transfected into AML12 cells, then the hypoxia/reoxygenation model was established and cells were collected to detect the expression of NOD1, AIM2 and active-Caspase-1.
Results
The ALT and AST levels in IR group were significantly higher than those in sham group, and peaked at IR 12-h subgroup (P<0.05). HE staining showed that hepatic injury was the most severe at 12 h after reperfusion. TUNEL results showed that the number of apoptotic cells was the greated at 12 h after reperfusion. Western blotting showed that NOD1 protein expression was highest at 12 h after reperfusion. With the prolongation of reperfusion time, the expression of AIM2 and active-Caspase-1 gradually increased, and that of pro-Caspase-1 gradually decreased. The expression of NOD1, AIM2 and active-Caspase-1 decreased after transfection of NOD1 siRNA into AML12 cells.
Conclusions
NOD1 promotes liver ischemia-reperfusion injury, which may be related to NOD1 promoting liver injury by activating pyroptosis.
Key words:
Reperfusion injury; Liver; Pyroptosis
Owing to oversights during the production process, a pair of errors were introduced into the above article. First, in the Materials and methods section, Animals and cell lines subsection, on p. 1332, line 12, the sentence starting here should have read as: 'The non‑tumorigenic mouse hepatocyte alpha mouse liver 12 (AML12) cell line was purchased from the Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China)' (i.e., the abbreviation AML12 was expanded incorrectly). Secondly, the bottom row of data was accidentally omitted from Table I (pertaining to the Suzuki score for liver ischemia/reperfusion injury assessed to be severe).The fifth data row in the Table should have read, from left to right, as follows: Numerical assessment, '4'; Congestion, 'Severe (>60%)'; Vacuolization, 'Severe (>60%)'; and Necrosis, 'Severe (>60%)'.We regret that these errors were introduced into the published version of this paper, and apologize to the authors and the readership for the inconvenience caused. [the original article was published in International Journal of Molecular Medicine 43: 1331‑1342, 2019; DOI: 10.3892/ijmm.2019.4077].
Liver ischemia/reperfusion injury (LIRI) is a common complication of liver surgery, and affects liver function post‑transplantation. However, the precise mechanism underlying LIRI has not yet been completely elucidated. Previous studies have demonstrated the involvement of a number of microRNAs (miRNAs/miRs) in liver pathophysiology. The objective of the present study was to determine the potential function and mechanism of miR‑101‑mediated regulation of autophagy in LIRI. Compared with the sham‑treated group, a significant decrease in miR‑101 and mechanistic target of rapamycin (mTOR) expression levels following ischemia/reperfusion (IR) were observed, along with an increased number of autophagosomes (P<0.001). The exogenous overexpression of miR‑101 has been demonstrated to inhibit autophagy during the LIRI response and the levels of mTOR and phosphorylated (p)‑mTOR expression are correspondingly elevated. However, compared with the miR‑NC group, miR‑101 silencing was associated with reduced mTOR and p‑mTOR levels and increased autophagy, as indicated by the gradual increase in the levels of the microtubule‑associated protein 1 light II (LC3II). The peak levels of LC3II were observed 12 h subsequent to reperfusion, which coincided with the lowest levels of miR‑101. In addition, inhibition of autophagy by 3‑methyladenine significant enhanced the protective effect of miR‑101 against LIRI, compared with the IR group (P<0.001). Altogether, miR‑101 attenuates LIRI by inhibiting autophagy via activating the mTOR pathway.
[Objective]Observation on the clinical curative effects of patients with Diabetic Peripheral Neuropathy(DPN)treated by medica Xuebidan mixture.[Methods]Designed with prospective methods,40 patients was given Xuebidan mixture(preparations in Dandong Hospital of TCM)60ml,qd,before breakfast and dinner based on regular blood glucose and complications control. Make Sure all patients never take other medicine for DPN,reflect and deal with the adverse reactions and disease changes.One period of treatment was 2 weeks.Observation on physical signs,Clinical score and adverse reaction,evaluated the clinical curative ef fect and safety after 2 two periods.[Results]Clinical cured:2 cases,significantly effectiveness:23 cases, effectiveness:14 cases,ineffectiveness:1 case,the total effective rate(clinical cured rate + significantly effective rate + effective rate)was 97.5%.[Conclusions]Xuebidan Mixture had better curative effect and no side effect for DPN,it's well worth to spread in clinical.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)
