Evidence of an anti-apoptotic effect of qinghuobaiduyin on intestinal mucosa following burn injury.

Burn injuries are common in wartime and in times of peace. The prevention and therapy of ischemia-reperfusion injury to the organs, in particular the intestine, during the burn shock and recovery process has become a popular yet challenging area of research. Studies concerning the apoptosis of the cells of the burned intestinal mucosa have gained considerable attention. Qinghuobaiduyin (QHBDY) is a traditional Chinese medicine that has been used as a clinical prescription since 1995 to treat burn patients due to its opsonization function in the immune system and favorable clinical therapeutic effect. The aim of this study was to investigate the effect of QHBDY on the apoptosis of intestinal mucosa following burn injury. An animal model was constructed comprising severely burned rats that were treated with various dosages of QHBDY. Tissues from the small intestine were collected to investigate the apoptosis rate by TUNEL assay and the protein expression levels of heat shock protein 70 (Hsp70) and caspase-3 by immunohistochemistry. In addition, IEC-18 cells treated with QHBDY and burn serum were investigated. The cell apoptosis rate was analyzed by flow cytometry (FCM), the protein expression levels of Hsp70 were measured by western blot analysis and caspase-3 activity was analyzed by a colorimetric assay. The results showed that in animal experiments, compared with the burned group, the apoptosis rates in the treatment group was decreased, the protein expression level of Hsp70 was increased while Caspase-3 was decreased. In cell experiments, after treatment with QHBDY, the cell apoptosis rate was lower than that of the burn serum group. In addition, Hsp70 protein expression was upregulated and caspase-3 activity was decreased. QHBDY may play an important role in the prevention of apoptosis at the whole animal and cellular levels.

Saikosaponin A mediates the inflammatory response by inhibiting the MAPK and NF-κB pathways in LPS-stimulated RAW 264.7 cells.

Saikosaponin A (SSA) is a major triterpenoid saponin isolated from Radix bupleuri (RB), a widely used Chinese traditional medicine to treat various inflammation-related diseases. The aim of this study was to investigate the anti-inflammatory activity, as well as the molecular mechanism of SSA in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In this study, we demonstrated that SSA markedly inhibits the expression of certain immune-related cytotoxic factors, including cyclooxygenase-2 (COX-2) and inducible nitric-oxide synthase (iNOS), as well as pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6. It also significantly upregulates the expression of IL-10, an important anti-inflammatory cytokine, suggesting its anti-inflammatory activity in LPS-stimulated macrophages. We further demonstrated that SSA inhibits the activation of the nuclear factor-κB (NF-κB) signaling pathway by suppressing the phosphorylation of inhibitory NF-κB inhibitor α (IκBα) and thus holding p65 NF-κB in the cytoplasm to prevent its translocation to the nucleus. In addition, SSA also inhibits the mitogen-activated protein kinase (MAPK) signaling pathway by downregulating the phosphorylation of p38 MAPK, c-Jun N-terminal kinase (c-JNK) and extracellular signal-regulated kinase (ERK), the three key components of the MAPK family. In conclusion, our study demonstrates that SSA has an anti-inflammatory effect by regulating inflammatory mediators and suppressing the MAPK and NF-κB signaling pathways in LPS-stimulated RAW 264.7 cells.

[Effect of qinghuobaiduyin on the expression of high mobility group box chromosomal protein 1 in macrophage].

OBJECTIVE: To observe the expression of high mobility group box chromosomal protein 1(HMGB1) in RAW264.7 macrophages after interfering with burning serum and qinghuobaidu-yin (QHBDY), and to find out the endogenous protection mechanism of QHBDY resisting inflammation reaction. METHODS: RT-PCR was used to detect the expression of HMGB1 in RAW264.7 macrophages after interfering RAW264.7 macrophages with normal SD rat serum, burning SD rat serum, and QHBDY feeding SD rat serum. RESULTS: Small quantity of HMGB1 mRNA was expressed in RAW264.7. The expression of HMGB1 mRNA fluctuated around the standard level after interfering with normal serum of SD rats. The expression of HMGB1 mRNA rose at 3 h, and then decreased to the standard level; at 18 h, it rose rapidly; at 36 h, it reached the peak; and at 48 h, it remained at the high level after interfering with burning serum. The expression of HMGB1 mRNA increased at 3 h, and then decreased to the standard level. At 24 h, it started to rise after interfering with herb serum, and was lower than that of; the burning serum group (P<0.05). CONCLUSION: Burning serum can increase the expression of HMGB1 mRNA in RAW264.7. QHBDY can decrease the high expression of HMGB1 mRNA in RAW264.7 caused by burning serum.

[Effect of Chinese traditional medicine mixture on inflammatory response in rats with severe burn].

OBJECTIVE: To investigate the regulatory effect of Chinese traditional medicine mixture (CTMM) on inflammatory response in rats with severe burn. METHODS: One hundred and ten rats were randomly divided into 3 groups:scalded rats inflicted by 30% III degree scald were treated with CTMM and SD-Ag (CTMM group), scalded rats inflicted by 30% III degree scald were treated with SD-Ag alone (scalded group), and healthy rats were treated with SD-Ag (normal group). The serum contents of TNF-alpha, IL-beta, IL-6, IL-8, IL-4, and IL-10 in rats in the 3 groups were dynamically monitored. RESULTS: The serum contents of TNF-alpha, IL-1beta, IL-6, IL-8, IL4, and IL-10 evidently increased in both the CTMM and scalded groups. But the contents of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6, and IL-8) in the CTMM group were much lower than those in the scalded group. However, the contents of anti-inflammatory cytokines (IL-4 and IL-10) in the CTMM group were much higher than those in the scalded group. CONCLUSION: CTMM has double-way regulatory effect on the inflammatory response in rats with severe burn.

[Effect of Chinese drugs mixture on immune function of patients with extremely severe burn].

OBJECTIVE: To study the regulatory effect of Qinghuo Baidu Yin (QHBDY, a mixture prepared with Chinese drugs) on immune function of patients with extremely severe burn (ESB). METHODS: Thirty patients with ESB were divided into two groups, conventional therapy was given to both groups, but QHBDY was given to the treated group additionally. Immunological indices, including peripheral blood T-lymphocyte subsets, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) were determined 3 weeks after treatment to evaluate and compare the therapeutical effect in the two groups. RESULTS: Compared with the control group, CD3, CD4, CD4/CD8, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) levels were markedly decreased in degree, and recovered earlier and quicker, with CD8 increased mildly (P < 0.01) and turned back more quickly. And so did the parameters of the treated group in comparing with that of the control group at anytime (P < 0.05 or P < 0.01). CONCLUSION: Chinese drugs mixture shows the regulatory effect on both cellular and humoral immune function in patients with ESB.