[Changing laws of serum high mobility group box 1 protein in septic rats and the intervention effect of xuebijing].

OBJECTIVE: To investigate the changing laws of serum high mobility group box 1 protein (HMGB1) in septic rats and intervention effect of Xuebijing on it. METHODS: Lipopolysaccharide (LPS) (5 mg/kg BW) was intravenously injected into the tail vein of healthy male Wistar rats to prepare the sepsis rat model. In Experiment 1: 50 Wistar rats were randomly divided into three groups, i.e., the normal group (A, n=10); the LPS model group (B, n=10), the LPS +Xuebijing treatment group (C, n=30). Rats in the C group were further divided into three subgroups, i.e., 2 h before LPS injection (group C1), 2 h after LPS injection (group C2), and 8 h after LPS injection (group C3), 10 in each group. Blood samples were collected from the caudal vein to detect serum HMGB1 levels by Western blot at 4, 12, 24, 48, and 72 h after LPS injection. Experiment 2: 30 Wistar rats were equally divided into the LPS model group (D) and the LPS + Xuebijing treatment group (E), 15 in each group. They were treated as rats in the B group and the C1 group respectively. Five rats were sacrificed at 12, 24, and 48 h after LPS injection in the two groups. Blood as well as the tissue samples were harvested to measure such indices as ALT, AST, Cr, and BUN, as well as pathological changes of liver, lung, and kidney. RESULTS: (1) Compared with the A group, serum HMGB1 levels were higher at various time points in the B group (P < 0.05). Compared with the B group, serum HMGB1 levels at 12,24,48, and 72 h decreased in the C1, C2, and C3 groups. Besides, the decrease was more obvious at 24 h and 48 h.The decrement in the C3 group was less than that in the C1 and C2 groups (P < 0.05). (2) In the D group, ALT, AST, Cr, and BUN were significantly higher than those in the A group and reached the peak at 24 h (P < 0.05). Compared with the E group, AST, Cr, and BUN at 24 and 48 h, and ALT at each time point decreased significantly in the E group (P < 0.05). (3)The results of pathological section of liver, lung, and kidney showed local congestion and hemorrhage, cell edema/necrosis/degeneration, infiltration of inflammatory cells, damage of characteristic structures and so on; particularly serious lesion occurred at 24 and 48 h in the D group. The microscopic lesion was obviously alleviated in the E group than in the D group at corresponding time points. CONCLUSIONS: The serum HMGB1 levels increased in septic rats, with late occurrence of peak value and longer duration of the high value. HMGB1 played an important role in excessive inflammatory response and multiple organ dysfunction. Xuebijing could reduce the serum levels of HMGB1, improve biochemical parameters, and attenuate severe inflammatory response of liver, lung, and kidney tissues in septic rats. Besides, the earlier use, the better effect obtained.

Xuebijing Protects Rats from Sepsis Challenged with Acinetobacter baumannii by Promoting Annexin A1 Expression and Inhibiting Proinflammatory Cytokines Secretion.

Xuebijing (XBJ) injection is a herbal medicine that has been widely used in the treatment of sepsis in China; however, its role in the development and progression of Acinetobacter baumannii sepsis and the underlying mechanisms remain uninvestigated. In the present study, fifty-four male Wistar rats were randomly assigned to normal-control group, sepsis-control group, and sepsis + XBJ group, each containing three subgroups of different treatment time periods (6, 12, and 24 hrs following injection, resp.). The sepsis model was established by intraperitoneal injection of A. baumannii ATCC 19606. For XBJ treatment, 4 mL/kg XBJ was administrated simultaneously by intravenous injection through caudal vein every 12 hrs. All animals demonstrated ill state, obvious intestinal dysfunction, histopathological lung damages, and overactive inflammatory responses after A. baumannii infection, and these events could be partially reversed by XBJ treatment from the beginning of infection. XBJ induced an increase in the expression of anti-inflammatory mediator annexin A1; however, two proinflammatory cytokines, interleukin-8 (IL-8) and tumor necrosis factor- α (TNF- α ), were decreased at the each monitored time point. These findings suggested that XBJ via its cytokine-mediated anti-inflammatory effects might have a potential role in preventing the progression of A. baumannii infection to sepsis by early administration.

[The effect of Xuebijing on liver translationally controlled tumor protein expression in Acinetobacter baumannii sepsis rats].

OBJECTIVE: To study the effect of Xuebijing on liver expressing translationally controlled tumor protein (TCTP) in Acinetobacter baumannii sepsis rats. METHODS: Among 42 healthy adult male Wistar rats of clean grade, 6 rats were randomly selected as the control group, others were randomly divided into two groups by the method of random digits table: sepsis group(n=18), Xuebijing group(n=18). Sepsis model was established through intraperitoneal injecting Acinetobacter baumannii suspension, and the Xuebijing injection was administrated through caudal vein 30 minutes later in Xuebijing group. After making model for 6, 12 and 24 hours, 6 rats were randomly selected from sepsis group and Xuebijing group, and then the rats were sacrificed, liver tissue samples were extracted for hematoxylin and eosin (HE) staining. Pathological changes of the liver were observed, and immunohistochemical analysis of liver tissue TCTP expression positive cells and the expression of TCTP in liver cells were detected by Western blotting method. RESULTS: HE staining of liver indicated that it was inflammatory injured in sepsis group, and inflammation decreased in Xuebijing group. Immunohistochemistry results showed that, compared with the control group, TCTP positive cells expression score at 6, 12 and 24 hours in sepsis group were significantly increased (7.33±0.82, 10.67±1.21, 7.67±1.21 vs. 2.50±1.05, all P<0.05). Compared with sepsis group, liver tissue TCTP positive cells expression score at 6, 12 and 24 hours in Xuebijing group (5.83±0.75, 7.50±1.05, 5.67±1.37) were significantly decreased (all P<0.05). Western blotting results showed that, compared with the control group, TCTP expression at 6, 12 and 24 hours in sepsis group were significantly increased (1.94±0.59, 3.20±0.72, 1.96±0.55 vs. 0.93±0.24, all P<0.05); compared with sepsis group, TCTP expression at 6, 12 and 24 hours in Xuebijing group (1.38±0.36, 2.03±0.49, 1.30±0.30) were significantly decreased (all P<0.05). CONCLUSIONS: Xuebijing can reduce inflammatory injury in liver of rats with Acinetobacter baumannii sepsis, and its mechanism may be associated with reduced hepatic cells expressed TCTP.