Effects of Shen-Fu Injection on the Expression of T-Cell-Specific Transcription Factors T-bet/Gata-3 in Porcine Postresuscitation Lung Injury

Abstract

Shen-Fu injection (SFI) derived from the ancient traditional Chinese medicine. In this study, the effects of SFI on the expression of T-bet/GATA-3 and its potential mechanisms causing the shift of T cells from Th2 to Th1 on postresuscitation lung injury were examined in a porcine model of cardiac arrest. 30 pigs were randomly divided into SHAM (n = 6) and three return of spontaneous circulation (ROSC) groups (n = 8 per group); 24 pigs were subjected to 8 min of electrically induced cardiac arrest and 2 min of basic life support, which received central venous injection of Shen-Fu (SFI), epinephrine (EP) or saline (SA). After successful ROSC, 18 surviving pigs were sacrificed at 24 h after ROSC (n = 6 per group). The levels of serum and lung tissue interleukin (IL)-4 and interferon (IFN)- γ were measured by ELISA, and the protein and mRNA levels of GATA-3 and T-bet in the lung tissue were determined by western blotting and quantitative real-time polymerase chain reaction, respectively. Compared with the EP and SA groups, SFI treatment reduced the levels of IL-4 (P < 0.05), increased levels of IFN- γ (P < 0.05), and induced T-bet mRNA upregulation and GATA-3 mRNA downregulation (P < 0.05). SFI attenuated lung injury and regulated lung immune disorders. Therefore, SFI could protect postresuscitation lung injury by modulating a Th1/Th2 imbalance.

Figure 1

Lung histopathology photographs. (a)–(d) Light microscope images (HE, ×200); (e)–(h) electron microscope images; (a) and (e) photographs from the sham-operated group; (b), (c), (f), and (g) photographs from the EP group; (d) and (h) photographs from Shen-Fu injection group. (b) and (c) Pulmonary arterioles contain erythrocytes, oedema fluid, and inflammatory cells (arrows). (d) Congestion, inflammatory cell infiltration and edema of the lung tissue were considerably alleviated (arrows). (e) normal type II epithelial cells structure was observed in the SHAM group (arrows). (f) Type II epithelial cells are heavily injured, exhibiting necrosis and disintegration (arrows). (g) The structure of the lamellar bodies in type II epithelial cell presents vacuolar change (arrows) inside the cell. (h) The lamellar body of type II epithelial cells shows minor damage in SFI group (arrows).

Figure 2

ELISA analysis of (a)–(c) IL-4, IFN-γ, and IFN-γ/IL-4 levels in the serum; (d)–(f) IL-4, IFN-γ, and IFN-γ/IL-4 levels in the lung tissue: significant difference (*P < 0.05; **P < 0.01 versus SHAM) between the SHAM group, SA, EP, and SFI group (n = 6), ^▲ P < 0.05, ^▲▲ P < 0.01 versus EP and SA.

Figure 3

ELISA analysis of IL-4, TNF-α, and IFN-γ levels in the lung tissue: (a) western blots of GATA-3 and T-bet protein expressions in the lung tissue at 24 h after ROSC. (b) Western blots of quantification of GATA-3 and T-bet protein levels. (c) mRNA expressions of GATA-3 and T-bet in the lung tissue at 24 h after ROSC. The values represent mean ± SE (n = 6) *P < 0.05 and **P < 0.01 versus SHAM; ^▲ P < 0.05, ^▲▲ P < 0.01 versus EP and SA. (one-way repeated-measures ANOVA).