XBJ Group Research Articles

The mechanism of Xuebijing injection in preventing and treating lung injury induced by cardiopulmonary bypass by regulating the apoptosis of alveolar polymorphonuclear neutrophil

To investigate the protective effect of Xuebijing injection on acute lung injury (ALI) associated with cardiopulmonary bypass (CPB) by regulating the apoptosis of polymorphonuclear neutrophils (PMN). Thirty male Sprague-Dawley (SD) rats were randomly divided into sham operation group (Sham group), CPB model group (CPB group) and Xuebijing pretreatment group (XBJ group) according to the random number table method, with 10 rats in each group. Rats in the CPB group and XBJ group undergoing CPB procedures for 60 minutes. Rats in the Sham group did not undergo CPB. Rats in the XBJ group received intraperitoneal injection of 4 mL/kg Xuebijing injection 2 hours before CPB. Rats in the Sham group and CPB group were injected with an equal amount of normal saline. 4 hours after CPB, arterial blood was collected for blood gas analysis to calculate respiratory index (RI), and lung tissue of rats was collected for determination of lung index (LI) and pulmonary water containing rate. PMN in bronchoalveolar lavage fluid (BALF) were collected and the activity of caspase-3 was detected. The apoptosis rate was detected by flow cytometry. The expressions of microRNA-142-3p (miR-142-3p) and FoxO1 mRNA were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The protein expression of FoxO1 was detected by Western blotting. In addition, HL-60 cells were divided into control oligonucleotide transfection group, miR-142-3p mimics transfection group, and miR-142-3p inhibitor transfection group. After 48 hours of transfection, the activity of miR-142-3p binding to FoxO1 was detected using dual luciferase reporter genes. Compared with Sham group, RI, LI and pulmonary water containing rate were significantly increased in CPB group. The caspase-3 activity and apoptosis rate of PMN obtained from BALF were significantly decreased, the expression of miR-142-3p was decreased, and the expression of FoxO1 protein was increased. However, compared with CPB group, RI, LI and pulmonary water containing rate were significantly decreased in XBJ group [RI: 0.281±0.066 vs. 0.379±0.071, LI: 4.50±0.26 vs. 5.71±0.42, pulmonary water containing rate: (80.31±32.50)% vs. (84.59±3.41)%, all P < 0.01]. The caspase-3 activity and apoptosis rate of PMN obtained from BALF were significantly increased [caspase-3 activity: 0.350±0.021 vs. 0.210±0.014, apoptosis rate: (15.490±1.382)% vs. (8.700±0.701)%, both P < 0.01], the expression of miR-142-3p was significantly up-regulated (2-ΔΔCt: 2.61±0.17 vs. 0.62±0.05, P < 0.01), and the protein expression of FoxO1 was decreased [FoxO1/GAPDH (relative expression level): 0.81±0.04 vs. 1.22±0.06, P < 0.01]. However, there was no statistically significant difference in FoxO1 mRNA expression among the three groups. The bioinformatics analysis results showed that miR-142-3p can bind to the FoxO1 3'untranslated region (3'UTR). In HL-60 cells, compared with control oligonucleotide transfection group, the transfection of miR-142-3p mimics could reduce the expression of FoxO1 protein [FoxO1/GAPDH (relative expression level): 0.48±0.06 vs. 1.00±0.05, P < 0.01], however, the transfection of miR-142-3p inhibitor increased the expression of FoxO1 protein [FoxO1/GAPDH (relative expression level): 1.37±0.21 vs. 1.00±0.05, P < 0.05]. But, transfection with miR-142-3p mimics or inhibitor had no effect on FoxO1 mRNA expression. The luciferase reporter gene showed that miR-142-3p could bind to the FoxO1 3'UTR to inhibit FoxO1 expression. Xuebijing injection may promote the apoptosis of pulmonary alveolar PMN through the miR-142-3p/FoxO1 axis, and play a role in the prevention and treatment of CPB-induced ALI.

Positive effects of Xuebijing injection on intestinal microbiota and metabolite spectrum in septic rats

To explore the effect of Xuebijing injection on inflammation in sepsis by regulating intestinal microbiota and its metabolites. A total of 45 male Sprague-Dawley (SD) rats were randomly divided into Sham operation group (Sham group), cecal ligation and perforation (CLP) induced sepsis group (CLP group), and Xuebijing intervention group (XBJ group, 4 mL/kg Xuebijing injection was injected intraperitoneally at 1 hour after CLP), with 15 rats in each group. The survival of rats was observed at 24 hours after operation and sacrificed. Feces were collected for 16S rRNA gene sequencing and liquid chromatography-mass spectrometry (LC-MS) analysis. At 24 hours after operation, all rats in the Sham group survived, the mortality of rats in the XBJ group was lower than that in the CLP group [47% (7/15) vs. 60% (9/15), P > 0.05]. Compared with the Sham group, the diversity of gut microbiota in the CLP group decreased, the dominant flora changed, and the abundance of inflammation-related flora increased. Xuebijing improved the changes in gut microbiota caused by sepsis, and α diversity showed an increasing trend (Ace index: 406.0±22.5 vs. 363.2±38.2, Chao1 index: 409.7±21.8 vs. 362.4±42.5, both P > 0.05). Restrictive constrained principal coordinate analysis (cPCoA) showed a high similarity in gut microbiota among the same group of rats. The CLP group was dominated by Bacteroidetes, while the Sham and XBJ groups were dominated by Firmicutes. In addition, compared with the CLP group, Xuebijing treatment increased the abundance of beneficial bacteria in septic rats, such as Verrucomicrobia, Akkermansia and Lactobacillus. LC-MS and orthogonal partial least squares discriminant analysis (OPLS-DA) showed that there were 12 main differential metabolites among the three groups, and there were certain correlations between these metabolites, which were related to amino acid and lipid metabolism. Correlation analysis showed a significant correlation between changes in metabolites and microbial communities. Xuebijing can improve the survival rate of septic rats, regulate the composition of intestinal flora and related metabolites, which provides a new pathophysiological mechanism for Xuebijing in the treatment of sepsis.

Xuebijing injection protects against sepsis-induced myocardial injury by regulating apoptosis and autophagy via mediation of PI3K/AKT/mTOR signaling pathway in rats.

Apoptosis and autophagy are significant factors of sepsis induced myocardial injury (SIMI). XBJ improves SIMI by PI3K/AKT/mTOR pathway. Present study is devised to explore the protective mechanism of XBJ in continuous treatment of SIMI caused by CLP. Rat survival was first recorded within 7 days. Rats were randomly assigned to three groups: Sham group, CLP group, and XBJ group. The animals in each group were divided into 12 h group, 1 d, 2 d, 3 d and 5 d according to the administration time of 12 hours, 1 day, 2 days, 3 days or 5 days, respectively. Echocardiography, myocardial injury markers and H&E staining were used to detect cardiac function and injury. IL-1β, IL-6 and TNF-α in serum were measured using ELISA kits. Cardiomyocyte apoptosis was assayed by TUNEL staining. Apoptosis and autophagy related proteins regulated by the PI3K/AKT/mTOR signaling pathway were tested using western blot. XBJ increased the survival rate in CLP-induced septic Rat. First of all, the results of echocardiography, H&E staining and myocardial injury markers (cTnI, CK, and LDH levels) showed that XBJ could effectively improve the myocardial injury caused by CLP with the increase of treatment time. Moreover, XBJ significantly decreased the levels of serum inflammatory cytokines IL-1β, IL-6 and TNF-α in SIMI rats. Meanwhile, XBJ downregulated the expression of apoptosis-related proteins Bax, Cleaved-Caspase 3, Cleaved-Caspase 9, Cytochrome C and Cleaved-PARP, while upregulated the protein levels of Bcl-2 in SIMI rats. And, XBJ upregulated the expression of autophagy related protein Beclin-1 and LC3-II/LC3-I ratio in SIMI rats, whereas downregulated the expression of P62. Finally, XBJ administration downregulated the phosphorylation levels of proteins PI3K, AKT and mTOR in SIMI rats. Our results showed that XBJ has a good protective effect on SIMI after continuous treatment, and it was speculated that it might be through inhibiting apoptosis and promoting autophagy via, at least partially, activating PI3K/AKT/mTOR pathway in the early stage of sepsis, as well as promoting apoptosis and inhibiting autophagy via suppressing PI3K/AKT/mTOR pathway in the late stage of sepsis.

To explore the preventive and therapeutic effects of Xuebijing injection on acute lung injury induced by cardiopulmonary bypass in rats by regulating the expression of microRNA-17-5p and its mechanism

To investigate the preventive effect of Xuebijing injection on acute lung injury induced by cardiopulmonary bypass (CPB) and the underlying mechanism. (1) In vivo experiment: 30 Sprague-Dawley (SD) rats were randomly divided into sham group, CPB group, Xuebijing pretreatment group (XBJ+CPB group) with 10 rats in each group. CPB model was reproduced in rats; and CPB was not performed in sham group, but only through arteriovenous puncture. In the XBJ+CPB group, 4 mL/kg Xuebijing injection was injected intraperitoneally 2 hours before CPB, sham group and CPB group were injected with equal volume of normal saline at the same time. The blood from femoral artery was analyzed 4 hours after operation, and the oxygenation index (PaO2/FiO2) was calculated. Then the rats were sacrificed to collect bronchoalveolar lavage fluid (BALF), and the lung permeability index (PPI) was calculated. The lung tissues were harvested, and the wet/dry weight ratio (W/D) of lung tissue was measured. The index of quantitative evaluation of alveolar injury (IQA) was measured. The levels of interleukins (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) in lung tissue and BALF were measured by enzyme-linked immunosorbent assay (ELISA). The content of malondialdehyde (MDA) and the activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in lung tissue were detected by biochemical method. The microRNA-17-5p (miR-17-5p) expression in lung tissue was determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). (2) In vitro experiments: type II alveolar epithelial cells (AEC II) were cultured in vitro, and they were randomly divided into control group (the cells were treated by preoperative serum of CPB in patients with ventricular septal defect), CPB group (the cells were treated by serum after CPB in patients), and XBJ+CPB group (Xuebijing injection 10 g/L+serum after CPB in patients). After 12 hours of culture in each group, the expression of miR-17-5p was detected by RT-qPCR. AEC II cells were transfected with miR-17-5p mimic, inhibitor or corresponding control oligonucleotide (negative control), respectively, to observe the effect of miR-17-5p on Xuebijing regulating CPB-induced apoptosis rate and caspase-3 activity. (1) In vivo experiment: compared with the sham group, the PPI, lung W/D ratio, IQA, and IL-1, IL-6, TNF-α in lung tissue and BALF, as well as MDA content and MPO activity in lung tissue were significantly increased, PaO2/FiO2 and SOD activity in lung tissue were significantly decreased. The parameters of the XBJ+CPB group were significantly improved, suggesting that Xuebijing pretreatment could improve CPB-induced ALI in rats. The expression of miR-17-5p in lung tissue of the CPB group was significantly down-regulated as compared with sham group (2-ΔΔCt: 0.48±0.13 vs. 1.00±0.11, P < 0.05); while the expression of miR-17-5p in the XBJ group was significantly up-regulated as compared with the CPB group (2-ΔΔCt: 1.37±0.09 vs. 0.48±0.13, P < 0.05), indicating that the improvement of Xuebijing injection on lung injury after CPB might be related to miR-17-5p. (2) In vitro experiment: the changes in miR-17-5p expression in each group of AEC II cells confirmed in vivo results. After transfection of miR-17-5p mimic, the apoptotic rate and caspase-3 activity of each group were significantly lower than those transfected with negative control, and the decrease was more significant in the XBJ+CPB group [apoptotic rate: (7.37±0.95)% vs. (12.60±1.90)%, caspase-3 (A value): 0.82±0.09 vs. 1.37±0.08, both P < 0.05]. After transfection of miR-17-5p inhibitor, the apoptotic rate and caspase-3 activity of each group were significantly more than those transfected with negative control [in the XBJ+CPB group: apoptotic rate was (16.30±1.86)% vs. (12.60±1.90)%, caspase-3 (A value) was 1.78±0.13 vs. 1.37±0.08, both P < 0.05]. This indicated that the apoptosis of AEC II cells cultured in serum after CPB was significantly reduced by miR-17-5p, and further reduced by the pretreatment with Xuebijing. Xuebiing injection can reduce the inflammatory reaction and oxidative stress of lung tissue in rats with ALI induced by CPB, and improve oxygenation. The mechanism may be related to up-regulation of miR-17-5p expression in AEC II cells and inhibition of apoptosis of AEC II cells.

Efficacy of Xuebijing Injection () on Cardiopulmonary Bypass-Associated Pulmonary Injury: A Prospective, Single-center, Randomized, Double Blinded Trial.

To evaluate the efficacy of Xuebijing Injection (, XBJ) on the lung injury induced by cardiopulmonary bypass (CPB). Fifty patients undergoing CPB were randomized to either the saline group or XBJ group according to a random number table (25 cases in each group). The patients in the saline group received saline and patients in XBJ group received XBJ at 12 h prior to the operation, at the beginning of the operation, and at 12 h after the second injection. The PaO2/FiO2 at extubation 3 days post-operation, duration of ventilation in the intensive care unit (ICU), and lengths of stay in the ICU and hospital were recorded. The levels of inflammatory mediators including interleukin (IL)-1β, IL-8, IL-10, and C-reactive protein (CRP) in bronchoalveolar lavage fluid (BALF) and plasma were measured. The neutrophil count and elastase neutrophil elastase in BALF were also measured. In addition, adverse events were monitored. The PaO2/FiO2 in the XBJ group was higher than that in the saline group from 12 to 72 h post-operation (all P<0.05). The blood levels of IL-1β, IL-8, and CRP in the XBJ group from 12 to 72 h were all significantly lower than those in the saline group (all P<0.05). In contrast, the level of the anti-inflammatory cytokine IL-10 was significantly higher in the XBJ group than in the saline group (P<0.05). In addition, 4 patients presented with atelectasis in the saline group and none in the XBJ group. Ten patients experienced mild acute respiratory distress syndrome (ARDS) during hospitalization, and 5 patients with mild ARDS were in the XBJ group (P<0.05). XBJ shows protective potential against lung injury in patients who undergo CPB surgery, possibly through the downregulation of inflammatory mediators, reduction in neutrophil infiltration, and upregulation of IL-10 (Trial registry: ChiCTR-TRC-14004628).

UHPLC-Q-TOF MS-Based Metabolic Analysis for the Therapeutic Efficacy of "Xuebijing Injection" against Sepsis-Induced Acute Lung Injury.

“Xuebijing Injection” (XBJ) is a traditional Chinese medicine and has been wildly used in the treatment of sepsis in China. However, few studies have reported the use of XBJ in sepsis with acute lung injury (ALI). This study aimed to investigate the therapeutic efficacy of XBJ against sepsis-induced ALI. Generally a total of 27 mice were equally randomized into three groups: a sham group was given saline before sham operation. A sepsis group received the cecal ligation and puncture (CLP) operation only. A sepsis+XBJ group, XBJ, was injected at 72, 48, and 24 h before CLP operation. The lung tissue was collected for UHPLC-Q-TOF/MS profiling analysis, biomarker identification, and pathway analysis. With the analysis of principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), forty-five purine, amino acid, and sphingolipid metabolites in lung tissues were identified as potential biomarkers of sepsis-induced ALI, among which 22 were reversed in the sepsis+XBJ group significantly. Conclusively, our results suggest that purine metabolic pathway, glutathione metabolic pathway, sphingomyelin metabolic pathway, arachidonic acid metabolic pathway, and phospholipid metabolic pathway may be the potential therapeutic pathways to overcome sepsis-induced acute lung injury and we provided the potential mechanisms of protective effects of XBJ against ALI.

Xuebijing Injection (血必净注射液) increases early survival rate by alleviating pulmonary vasopermeability in rats subjected to severe burns

To investigate the effects of Xuebijing Injection (, XBJ) on survival rate and pulmonary vasopermeability in a rat model of severe scald injury. Rats were divided into two experiments: experiment 1 was monitored for 12 h post-injury for survival analysis after severe burns; in experiment 2, rats were killed for determination of pulmonary vascular permeability and pro-inflflammatory mediators. In both experiments, rats were subject to third-degree 50% total body surface area (TBSA) burns or sham injury followed by XBJ or normal saline (NS) treatment. In addition, rat pulmonary microvascular endothelium cells (PMECs) were pretreated with either XBJ or phosphate buffer saline (PBS), and then subjected to sham serum or scald serum stimulation for 2 or 6 h, followed by transwell examination for the permeability of PMECs. Meanwhile, pro-inflflammatory mediators in PMECs culture supernatant were also investigated. The average survival time in the scald+XBJ group was 582.1±21.2 min, which was signifificantly longer than that in the scald + NS group (345.8±25.4 min, P<0.01). Plasma levels of tumor necrosis factor-alpha (TNF-α), E-selectin, interleukin-6 (IL-6), vascular permeability and water content of lung tissues were signifificantly increased in animals after severe burns (P<0.01). However, administration of XBJ signifificantly decreased these levels in plasma and lung tissue. In in vitro cell experiments, XBJ markedly attenuated permeability in PMECs monolayer and reduced the levels of TNF-α, IL-6 and soluble E-selectin after stimulation with scald serum (P<0.01). XBJ increases early survival rate by alleviating pulmonary vasopermeability and inhibiting pro-inflflammatory mediators in rats subjected to lethal scald injury. XBJ may be a potent drug in treatment of severe burns.

Effects of xuebijing injection on cardiac function and structure in rats with myocardial hypoxia/reoxygenation injury

To investigate the protective effects of xuebijing (XBJ, Traditional Chinese Medicine Complex) injection on cardiac function in rats with myocardial hypoxia/reoxygenation(H/R) injury. The isolated langendorff perfused rat heart model was established. One hundred and thirty SD rats were randomly divided into sham group, hypoxia/reoxygenation group, low dose XBJ(XBJL) group, middle dose XBJ(XBJM) group and high dose XBJ(XBJH) group. All groups except sham group were divided into three subgroups according to reoxygenation time(0.5 h,1 h, 2 h) (n=10). In sham group, left ventricular development pressure(LVDP), maximal rates of increase/decrease of the left ventricular pressure(±dp/dtmax), left ventricular pressure (LVP), and heart rates (HR) were recorded after 20 minutes balance perfusion. The creatine kinase-MB (CK-MB) in myocardium was detected by ELISA. In other groups, after 20 minutes balance perfusion, we perfused ThomasⅡto stop the hearts from beating for 30 minutes, then reperfused the K-H until hearts recover beating. The microstructure of myocardium was observed under light microscopy. LVDP, ±dp/dtmax, LVP and HR were continuously recorded in other four groups and the concentrations of CK-MB in myocardium were measured by ELISA at different time points after reoxygenation. Microstructure of myocardium in each group were observed under light microscopy. LVDP, ±dp/dtmax, LVP and HR of other groups were significantly lower than those of sham group(P<0.05). The levels of CK-MB were higher than that of sham group(P<0.05). LVDP, ±dp/dtmax, LVP and HR of XBJL, XBJM and XBJH groups were higher than those of I/R group at corresponding time points after reoxygenation(P<0.05). The levels of CK-MB were lower than that of I/R group(P<0.05) and the cardiac function was improved. The middle dose of XBJ had the best protective effect. Xuebijing injection can effectively improve cardiac function and structure in rats with myocardial hypoxia/reoxygenation injury, and middle dose of XBJ is the best.

Effects of Xuebijing injection on microcirculation in septic shock

BackgroundThis study was conducted to investigate whether Xuebijing injection can rectify the dysfunction of microcirculation in septic shock and assessed the microcirculatory parameters directly via orthogonal polarization spectral and software AVA 3.0. Material and methodsAnesthetized and mechanically ventilated beagle dogs were modeled for septic shock via lipopolysaccharide (LPS) intravenous injection. They were divided randomly into four groups, control group accepted operation only for jejunostomy and cystostomy; LPS group accepted operation and LPS intravenous injection; saline group accepted operation, LPS intravenous injection, and saline resuscitation; XBJ group accepted operation, LPS intravenous injection, saline resuscitation, and Xuebijing injection infusion. The MAP was recorded via right femoral artery catheterization, at the same time, the blood gas analysis was taken via that pathway at the set time points. ScvO2 was obtained via right jugular vein catheterization at the related time points. Microcirculatory parameters were recorded by orthogonal polarization spectral via the jejunum stoma, and the microcirculation image was analyzed via the software AVA 3.0 later. ResultsXuebijing injection improved microcirculation of the jejunum villus in canine model of septic shock induced by endotoxin, especially for the proportion of perfused vessels. ConclusionsBased on the adequate fluid resuscitation, Xuebijing injection plays a helpful role of improving microcirculation in septic shock.

Pretreatment with Xuebijing injection alleviates systemic inflammatory response induced by severe heat-stroke via ameliorating intestinal injury in rats

To observe the effect of Xuebijing injection pretreatment on systemic inflammatory response induced by severe heat-stroke, and to investigate the mechanism of alleviation of intestinal injury in rats. Thirty-six healthy adult male Wistar rats with grade SPF were randomly assigned into three groups with randomized number method, namely sham group, severe heat-stroke model group, and Xuebijing pretreatment group ( XBJ group ), with 12 rats in each group. The animals were placed in a pre-warm chamber [ temperature ( 40±2 ) centigrade, humidity ( 65±5 )% ] in order to induce typical heat-stroke. The duration of heat-stress was 60 minutes, while the animals in sham group were exposed to ambient temperature of 25 centigrade. Arterial blood samples were collected at the beginning and the end of heat-stress, the concentrations of tumor necrosis factor-α( TNF-α), interleukins ( IL-1β, IL-6 ), and lipopolysaccharide ( LPS ) in peripheral blood were determined by enzyme linked immunosorbent assay ( ELISA ). The intestinal tissues were harvested after heat-stress, and the pathological changes in intestine tissues were observed after hematoxylin-eosin ( HE ) staining and under optical microscope. The pathological injury scores were calculated. Immunohistochemistry was performed to determine inducible nitric oxide synthase ( iNOS ) expression in intestinal tissue. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling ( TUNEL ) staining. Western Blot was used to measure the tight junction protein occludin expression. The concentrations of TNF-α, IL-1β, IL-6 and LPS in blood of the rats after heat-stress in model group were significantly higher than those of sham group [ TNF-α ( μg/L ): 443.00±110.10 vs. 98.36±44.61, IL-1β ( μg/L ): 436.37±163.64 vs.64.24±16.15, IL-6 ( μg/L ): 342.70±92.42 vs. 54.40±13.22, LPS ( μg/L ): 0.68±0.22 vs. 0.09±0.02, all P < 0.01 ], but the levels of these parameters in XBJ group were significantly lower than those of model group [ TNF-α ( μg/L ): 340.45±68.57 vs. 443.00±110.10, IL-1β ( μg/L ): 191.33±82.78 vs. 436.37±163.64, IL-6 ( μg/L ): 192.21±37.89 vs. 342.70±92.42, LPS ( μg/L ): 0.43±0.17 vs. 0.68±0.22, all P < 0.01 ]. Infiltration of inflammatory cells, necrosis and hemorrhage in intestinal mucosa were found in the intestine of heat-stroke animals in model group. The pathological lesions in XBJ group were milder than those of model group, with a decreased pathological injury score compared with model group ( 2.10±1.15 vs. 3.20±0.67, P < 0.01 ). The expression of iNOS and apoptosis of cells in intestinal tissue in model group were increased compared with that of sham group, but they were significantly less marked in XBJ group compared with model group [ iNOS ( adjusted A value ): 0.32±0.15 vs. 0.74±0.17, apoptotic index: 0.23±0.08 vs. 0.56±0.07, both P < 0.01 ]. The order of expression for occludin protein from high to low was sham group, XBJ group and model group ( A value was 0.96±0.25, 0.62±0.20, 0.33±0.11, respectively ). Furthermore, there was significant difference in the expression of occludin protein between model group and both XBJ group and sham group ( both P < 0.01 ). Xuebijing injection alleviates inflammation and endotoxemia produced by severe heat-stroke in rats. The mechanism may be related to amelioration of oxidative injury, apoptosis, and dysfunction of tight junction protein occludin expression.

血必净对脓毒症早期大鼠下丘脑–垂体–肾上腺轴的调节

目的：探讨脓毒症早期大鼠血浆中促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ACTH)、皮质醇(CORT)水平的变化，明确脓毒症时大鼠下丘脑-垂体-肾上腺(HPA)轴的变化以及血必净对其的调节作用。方法：选用健康雄性SD大鼠40只，随机分为正常对照组(Normal group)，假手术组(Sham group)，模型组(Sepsis group)和血必净治疗组(XBJ group)四个组，采用改良的盲肠结扎穿孔法(CLP)制作SD大鼠脓毒症模型。观察CLP术后动物的临床表现，术后6小时处死动物，利用放射免疫法检测血浆中促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ACTH)、皮质醇(CORT)水平。结果：XBJ组与Sepsis组相比，血浆中CRH、ACTH、CORT水平显著降低(P 0.05)。结论：脓毒症早期大鼠HPA轴处于过度激活的状态，血浆中CRH、ACTH、CORT的水平是升高的；血必净治疗后能够降低血浆中CRH、ACTH和CORT的水平，调节HPA轴的过度激活状态，减轻脓毒症大鼠的感染中毒表现。 Objective: To investigate the level changes of corticotropin-releasing hormone (CRH), adren- ocorticotropic hormone (ACTH) and cortisol (CORT) level in plasma in septic rats, without and with the use of Xuebijing injection. The main purpose is to reveal whether Xuebijing can affect the hypothalamus-pituitary-adrenal (HPA) axis in septic rats. Methods: Forty male Sprague- Dawley rats were randomly divided into 4 groups: control group, sham group, saline group (NS group), Xuebijing for treatment group. The septic rat models were challenged by CLP. We observe the clinical manifestation of animal after CLP. We use the radio-immunity method to detect the levels of CRH, ACTH and CORT in plasma. Results: Compared with the Sepsis group, the level of CRH, ACTH and CORT in XBJ group was significantly decreased (P 0.05). Conclusion: In septic rats, the levels of the CRH, ACTH and CORT in plasma were increased, which indicate that HPA axis was more active. Xuebijing can decrease the levels of CRH, ACTH and CORT in plasma, downregulate overactivation of the HPA axis, improves the clinical manifestation.

血必净对脓毒症大鼠皮质酮及肾上腺组织GRmRNA水平的影响

目的：探讨在血必净干预下，脓毒症大鼠血浆皮质酮(CORT)水平以及肾上腺组织糖皮质激素受体(GR mRNA)水平的变化，明确血必净对脓毒症大鼠CORT及GR mRNA的调节作用。方法：选用健康雄性SD大鼠40只，随机分为正常对照组(Normal group)，假手术组(Sham group)，模型组(Sepsis group)和血必净治疗组(XBJ group)四个组，采用经典的盲肠结扎穿孔法(CLP)制作SD大鼠脓毒症模型。CLP术后6小时处死动物，利用放射免疫法检测血浆皮质酮(CORT)水平；RT-PCR法检测肾上腺组织GRmRNA水平。结果：Sepsis组血浆CORT含量(29.74 ± 1.65 ng/ml)明显高于Normal组(7.56 ± 0.89 ng/ml, P 0.05)；Sepsis组肾上腺组织GR mRNA含量(0.80 ± 0.11)明显低于Normal组(1.44 ± 0.07, P 0.05)。结论：脓毒症早期大鼠血浆CORT的水平是升高的，而肾上腺组织GRmRNA水平是较低的；血必净干预后能够降低血浆CORT的水平并且一定程度上能够提高肾上腺组织GRmRNA的水平，减轻脓毒症大鼠的过度应激状态，改善脓毒症大鼠的感染中毒症状。 Objective: To investigate the change of CORT and the level of GRmRNA of adrenal in CLP induced septic rats which were treated by Xuebijing, identify that Xuebijing can regulate the CORT and GRmRNA. Methods: 40 male rats were divided randomly into normal control group, sham group, saline group (Sepsis group), Xuebijing for treatment group. The model of sepsis was made by CLP. We observe the clinical manifestation of animal after CLP. We use the radio-immunity method to detect the levels of CORT; using the RT-PCR method to measure the level of GRmRNA in adrenal. Results: In Sepsis group, the plasma CORT concentration (29.74 ± 1.65 ng/ml) was significantly higher than that in the normal control group (7.56 ± 0.89 ng/ml, P 0.05). In Sepsis group, the level of the GRmRNA in the adrenal tissue (0.80 ± 0.11) was significantly lower than that in the normal control group (1.44 ± 0.07, P 0.05). Conclusion: In septic rats, the HPA axis was excessive activate. Xuebijing can decrease the level of CORT, increase the level of GRmRNA in adrenal, improve the excessive activate condition of HPA axis, and lessen the clinical manifestation.

Effects of Xuebijing on hypothalamus-pituitary-adrenal axis in early stage of septic rats

To explore the levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), cortisone (CORT) and ultrastructural abnormalities in hypothalamus, pituitary, adrenal specimens and potential effects of Xuebijing injection in septic rats. Sepsis was induced in adult male Sprague-Dawley rats by cecal ligation and puncture (CLP). A total of 40 rats were randomly divided into control group (n = 10), sham-operated group (n = 10), saline treatment group (NS, 4 ml/kg, n = 10), Xuebijing injection treatment group (XBJ, 4 ml/kg, n = 10). The histological abnormalities associated with sepsis were examined in hypothalamus, pituitary and adrenals. Radioimmunoassay (RIA) was used to detect the levels of CRH in hypothalamic tissue and the plasma levels of ACTH and CORT. Compared with normal control or sham group, the levels of CRH in hypothalamus tissue of CLP group and plasma levels of ACTH and CORT increased in early stage of sepsis. In NS group, the plasma CORT concentration ((30.66 ± 1.55) ng/ml) was significantly higher than that in normal control group ((7.63 ± 0.56) ng/ml, P < 0.01) and sham group ((11.85 ± 0.87) ng/ml, P < 0.01). In XBJ group, the plasma CORT concentration ((22.13 ± 1.49) ng/ml) was significantly lower than that in NS group (P < 0.01). There was no significant difference between normal control and sham groups (P > 0.05). In NS group, the plasma concentration of ATCH ((26.26 ± 1.63) pg/ml) was significantly higher than that in normal control group ((8.84 ± 1.39) pg/ml, P < 0.01) and sham group ((11.43 ± 0.47) pg/ml, P < 0.01). In XBJ group, the plasma concentration of ATCH ((22.13 ± 1.49) ng/ml) was significantly lower than that in NS group (P < 0.01). No significant difference existed between normal control and sham groups (P > 0.05). In NS group, the level of CRH in hypothalamus tissue ((101.92 ± 6.61) ng/ml) was significantly higher than that in normal control group ((61.65 ± 6.05) ng/ml, P < 0.05) and sham group ((66.65 ± 4.04) ng/ml, P < 0.05). In XBJ group, the concentration of CRH in hypothalamus tissue ((84.90 ± 2.54) ng/ml) was significantly lower than that in NS group (P < 0.05). There was no significant difference between normal control and sham groups (P > 0.05). The ultrastructures of hypothalamus, pituitary and adrenal changed. In CLP group, the ultrastructure of hypothalamus was as follows: rough endoplasmic reticulum expanded. There were degranulation and Golgi complex swelling. A large number of endocrine granules could be seen in ATCH cells in pituitary and a depletion of adrenal lipid droplets. Xuebijing injection can lessen the excessive activated status of hypothalamic-pituitary-adrenal axis.