Clinical and animal studies on the expression of HMGB1 in Candida albicans sepsis and its role

HMGB1 and sepsis

OBJECTIVE:
Infectious fungal infection has become a serious threat to human health and plays an important role in bacterial sepsis. High-mobility group protein B1 (high mobility group box 1, HMGB1) It is unclear whether it plays an important role in invasive fungal infections. Candida albicans is also the main pathogen of invasive fungal infection. In this study, we will study the clinical and animal experiments of Candida albicans sepsis and study the expression of HMGB1 in order to explore the role of HMGB1 in invasive candidiasis infection and provide new treatment for the prevention and treatment of invasive Candida albicans infection Target. The levels of HMGB1, tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (interleukin-6) and interleukin-6 (IL-6) were detected by ELISA and q RT-PCR in the peripheral blood of patients with Candida albicans. -6, IL-6) and HMGB1 m RNA were analyzed and correlated with clinical indexes. The animal model of invasive Candida albicans infection was established. The experimental group was divided into normal group, immunosuppressive group, invasive Candida albicans infection group and invasive Candida albicans infection + ethyl pyruvate (EP) group, Mice were sacrificed at 1, 3, 5 and 7 days after infection. Liver and kidney cucumber and c-reactive protein (CRP) were measured by peripheral blood to determine the severity of the mice. The levels of HMGB1, TNF-alpha and IL-6 in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). The expression of HMGB1 m RNA in peripheral blood, lung, liver and kidney was detected by RT-PCR. Blot and immunohistochemistry were used to detect the expression of HMGB1 in lung, liver and kidney.

Results:
1. The peripheral blood of patients with Candida albicans q RT-PCR showed that patients with severe sepsis infected with Candida albicans, severe sepsis without fungal infection and peripheral blood of patients with sepsis The expression of HMGB1 m RNA was higher than that of normal people (P <0.01), while those with Candida albicans infection and severe sepsis without fungal infection were higher than those of general sepsis (P

2. Candida albicans patients with sepsis peripheral blood ELISA results showed that Candida albicans infected with severe sepsis patients, without fungal infection in patients with severe sepsis and sepsis serum HMGB1, TNF-alpha And IL-6 expression were higher than normal; and Candida albicans infection and non-fungal infection with severe sepsis patients and higher than the general sepsis patients; and Candida albicans infected with severe sepsis patients The expression of HMGB1 and TNF-alpha in serum was significantly higher than that in patients with severe sepsis without fungal infection, but the serum levels of IL-1 and IL-1 were significantly higher in patients with severe sepsis with Candida albicans infection and severe sepsis without fungal infection 6 expression was not significantly different. 3.Spearman correlation analysis showed that HMGB1 and procalcitonin (PCT), CRP and 1,3-beta-D-glucan (1,3-beta-D) in peripheral blood of patients with Candida albicans (R = 0.093, P <0.05), and there was no correlation (r = 0.093, P <0.05) with white blood cell (WBC) (r = 0.093, P <0.05). In mice, intraperitoneal injection of cyclophosphamide, the blood of white blood cells, red blood cells, hemoglobin and platelets were significantly reduced, of which the most significant reduction in leukopenia, indicating that immunosuppressive mouse model was successfully constructed. According to the survival curves of different concentrations of mice, the optimal concentration of invasive Candida albicans infection was 2 × 105 cells / ml. 6. Lung, liver and kidney tissue culture and pathological results showed: lung, liver and kidney tissue were Candida albicans growth, indicating that invasive candidiasis infection model was successfully constructed. Compared with the normal group, the body weight of the immunosuppressive group, the invasive Candida albicans infection group and the EP group was significantly decreased on the 4th day after immunosuppression (P <0.05). Compared with the immunosuppressive group, Candida albicans infection group and EP group began to lose weight on the 2nd day after infection (P <0.05). Compared with EP group, the mice infected with Candida albicans infection group decreased significantly at the third day after infection (P0 .05). The mortality rate of mice in invasive Candida albicans infection group was 33.33%, and the death rate of EP group was 13.33%. Compared with invasive Candida albicans infection group, EP could significantly reduce the mortality rate of mice. The difference was statistically significant (P <0.05). 9. Biochemical and pathological results showed that compared with the normal group and immunosuppressive group, invasive lung Candida infection group mice lung, liver and kidney tissue was impaired, CRP was significantly increased; and invasive Candida albicans Compared with the infection group, the damage of lung, liver and kidney tissue in EP group was significantly reduced and CRP was significantly decreased. The results of RT-PCR in peripheral blood, lung, liver and kidney tissues showed that compared with the normal and immunosuppressive groups, the peripheral blood, lung, liver and kidney tissues of the invasive Candida albicans infection group and the EP group (P

1. Immunohistochemical results of lung, liver and kidney showed that the expression of HMGB1 in the lung tissue of the infected Candida albicans infection group was significantly increased after the infection with Candida albicans, mainly in the nucleus; the expression of HMGB1 in the nucleus of the liver. The expression of HMGB1 in the cytoplasm of the liver cells was significantly increased with the prolongation of the number of days of infection. The expression of HMGB1 in the nucleus and cytoplasm of the renal tissue was significantly increased. Compared with the invasive Candida albicans infection group, EP can not only inhibit the expression of HMGB1 in the nucleus, but also inhibit the release of HMGB1 from the nucleus into the cytoplasm. The expression of HMGB1, TNF-alpha and IL-6 in peripheral blood of mice infected with Candida albicans and EP mice was significantly higher than that of the control group (P <0.05) The expression of HMGB1, TNF-alpha and IL-6 in peripheral blood of EP group was significantly lower than that of EP group.

2. HMGB1 in peripheral blood of patients with Candida albicans was positively correlated with PCT, CRP and BDG. According to other bacterial sepsis and the results of this study, we hypothesized that HMGB1 binds clinically different markers that help distinguish between bacterial sepsis and fungal sepsis. But this experiment to collect the sample size is small, this conjecture has yet to be further proved. The optimal infection concentration was 2 × 105 cells / ml, and the animal model of invasive Candida albicans infection was confirmed by tissue culture and histopathology. The expression of HMGB1m RNA and protein in peripheral blood, lung, liver and kidney of mice infected with Candida albicans was significantly higher than that of the control group. Compared with the invasive Candida albicans infection group, EP could significantly inhibit the expression and release of HMGB1, Reduce the degree of lung, liver and kidney damage, reduce the mortality of mice, suggesting that HMGB1 inhibitors may be invasive treatment of Candida albicans infection to provide a new target.

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