The impact of ulinastatin on lymphocyte apoptosis and autophagy in sepsis patients

Abstract

This study aimed to assess the influence of ulinastatin (UTI) on lymphocyte apoptosis and autophagy in sepsis patients, as well as its effect on inflammatory factors and vital organ function, with the goal of providing insights for improved clinical management of sepsis. A total of 40 sepsis patients were randomly assigned to the UTI group or the control group. The UTI group received standard treatment plus intravenous UTI, while the control group received standard treatment alone. Peripheral blood samples were collected at multiple time points for analysis of lymphocyte apoptosis, autophagy, inflammatory markers, and organ function. Various experimental techniques including Hoechst staining, transmission electron microscopy, and Western blot analysis were utilized to assess lymphocyte apoptosis, autophagy, and related protein expression levels. The study revealed that UTI treatment significantly inhibited lymphocyte apoptosis and promoted autophagy in sepsis patients. The levels of autophagy-related proteins LC3-II and Beclin-1 were substantially elevated, while the ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax was increased following UTI treatment. Furthermore, the levels of inflammatory markers IL-6, procalcitonin, and C-reactive protein were markedly reduced in the UTI group compared to the control group. Additionally, UTI treatment led to improved liver, kidney and cardiac function as evidenced by reduced levels of liver enzymes and creatinine, and cardiac enzymes. The findings of this study demonstrate that UTI exerts a protective effect on septic patients by inhibiting lymphocyte apoptosis, promoting autophagy, and attenuating systemic inflammation. Moreover, UTI treatment was associated with improved liver, kidney, and cardiac function in septic patients. These results contribute to a better understanding of the clinical management of sepsis and underscore the potential of UTI as a therapeutic intervention in septic patients.

Keywords: Autophagy; Inflammatory response; Lymphocyte apoptosis; Sepsis; Ulinastatin (UTI).

Fig. 1

Hoechst staining revealed a decrease in lymphocyte apoptosis in the study group. The cells indicated by the red border are apoptotic cells. (A) Apoptosis of lymphocytes was observed, which showed cell size reduction, wrinkled, nucleus shrinkage and dense staining; (B) The apoptotic lymphocytes increased from day 0; (C) The apoptotic lymphocytes were significantly higher than those on day 1, and were at the highest level in the experimental observation period; (D) The apoptotic lymphocytes were lower than that on day 3, but still higher than that on day 0; (E) Apoptosis of lymphocytes was observed, which showed cell size reduction, wrinkled, nucleus shrinkage and dense staining; (F) Apoptotic lymphocytes decreased from day 0; (G) The apoptotic lymphocytes were at the lowest level, with almost no apoptotic lymphocytes; (H) Although apoptotic lymphocytes were higher than those on day 3, they were still at a low level.

Fig. 2

Images of autophagy and apoptosis under transmission electron microscopy. The red arrow indicates cell apoptosis, while the yellow arrow indicates autophagy. (A) Lymphocyte swelling, disappearance of organelles, incomplete cell membrane, autophagosomes and apoptotic bodies can be observed; (B) The number of lymphocyte autophagosomes decreased compared with day 0, and cell apoptosis increased; (C) The lymphocyte has the least autophagosomes, almost no autophagosomes, and the apoptosis or necrosis of cells is the most serious; (D) There were more autophagosomes than on day 3, but the level was still low; (E) Lymphocyte swelling, disappearance of organelles, incomplete cell membrane, autophagosomes and apoptotic bodies can be observed; (F) The number of lymphocyte autophagosomes increased compared with day 0; (G) The lymphocytes had the most autophagosomes and no signs of apoptosis; (H) Although the level of autophagy was lower than that on day 3, autophagosomes were still observed.

Fig. 3

Immunoblotting images of autophagy and apoptosis proteins LC3II, Beclin-1, Bcl-2 and Bax alongside corresponding statistical charts. The bar chart displays the statistical results of the expression levels of relevant proteins in the control group and the study group at different time points (Day 0, Day 1, Day 3, and Day 7). *indicates p < 0.05 between the control and study groups; ***indicates p < 0.001 between the control and study groups; ****indicates p < 0.0001 between the control and study groups.

Fig. 4

Statistical chart of inflammatory factors IL-6, PCT, and CRP. The bar chart displays the statistical results of inflammatory factors in the control group and the study group at different time points (Day 0, Day 1, Day 3, and Day 7). *indicates p < 0.05 between the control and study groups; **indicates p < 0.01 between the control and study groups.

Fig. 5

Statistical chart of liver, kidney and cardiac function indicators: ALT, AST, Crea, CK-MB, and cTnI. The bar chart displays the statistical results of function indicators in the control group and the study group at different time points (Day 0, Day 1, Day 3, and Day 7). *indicates p < 0.05 between the control and study groups; **indicates p < 0.01 between the control and study groups; ***indicates p < 0.001 between the control and study groups.