HSP70 inhibits burn serum-induced apoptosis of cardiomyocytes via mitochondrial and membrane death receptor pathways.

The objective is to determine if Hsp70 is involved in the mechanism by which serum from burn rats induces apoptosis of cardiomyocytes. Cardiomyocytes in primary culture were assigned to the following groups: normal controls, normal rat serum, burn serum, burn serum plus empty vector control, and burn serum plus Hsp70 transfection. Each group, except normal controls, was cultured with the corresponding serum for 2 hours. The empty vector controls and the Hsp70 groups were transfected, respectively, with the empty vector or pcDNA3.1-Hsp70 recombinant plasmid 36 hours previously. Hsp70 protein levels were assessed by Western blot. Apoptotic cells were counted after Hoechst 33258 staining. Apoptosis of cardiomyocytes was also detected by the presence of "ladder" bands in agarose gels following electrophoresis of extracted DNA. Activation of caspase-3, -8, and -9 and Bid cleavage were assessed by Western blot. Hsp70 overexpression inhibited burn serum-induced apoptosis of cardiomyocytes; and burn serum-induced activation of caspases-3, -8, and -9; and Bid cleavage into tBid. Hsp70 inhibits burn serum-induced apoptosis by interfering with death receptor and mitochondrial pathways.

Influence of heat shock factor 1 gene transfection on the expression of inflammatory mediators in macrophages induced by burn serum / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of heat shock factor1 (HSF1) on gene expression of inflammatory mediators in RAW264.7 murine macrophage cells induced by burn serum.</p><p><b>METHODS</b>Sera were separated from blood of normal rats and rats with severe burns, and the recombinant vector pcDNA3. 1/HSF1 was constructed. RAW264.7 macrophages were divided into non-transfection group, vacant vector group (with burn and normal sera stimulation, respectively after vacant vector transfection) and recombinant vector group (with burn and normal sera stimulation, respectively after recombinant vector transfection). Some recombinant vector transfected macrophages without serum stimulation were prepared for the determination of HSF 1 expression with Western blotting. The mRNA expressions of TNF-alpha, HMGB 1 and IL-10 were determined with RT-PCR.</p><p><b>RESULTS</b>The cell line attained after recombinant vector transfection was comparatively stable,with partial activation of HSF 1. Burn sera markedly upregulated TNF-alpha, HMGB1 mRNA expression (0.910 +/- 0.100, 0.860 +/- 0.020, respectively), but downregulated IL-10 expression (0.430 +/- 0.010, respectively) in normal macrophages, while these genes maintained in a very low level in normal macrophages with normal serum stimulation . macrophages with recombinant vector transfection and burn serum stimulation could obviously inhibit the expression of TNF-alpha and HMGB 1, but enhance the IL-10 gene expression (0.130 +/- 0.100, 0.450 +/- 0.020 , 0.450 +/- 0.020, respectively )when compared with that with vacant vector transfection and burn serum stimulation (0.800 +/- 0.050, 0.880 +/- 0.030, 0.420 +/- 0.010, respectively).</p><p><b>CONCLUSION</b>HSF1 can inhibit the expression of some pro-inflammatory mediators in macrophages after a severe burns, indicating that appropriate upregulation of anti-inflammatory mediators might exert protective effects on the organism.</p>

Relationship between angiogenesis and expression of HO-1 of scar / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationshion between the angiogenesis of different kinds of scar and expression of HO-1.@*METHODS@#The expression of heme oxygenase-1 and vessel counted by CD34 of biopsies from different kinds of scars such as hypertrophic scar, keloid, surgical scar and normal skin of 24 cases was valued by immunochemical method, and the relationship was compared between them.@*RESULTS@#The vessel count of hypertrophic scar, keloid was significantly abundant compared with surgical scar or normal skin (P < 0.01). While the expression of HO-1 of hypertrophic scar, keloid was obviously higher than that in surgical scar or normal skin (P < 0.01), decreased from hypertrophic scar, keloid, surgical scar to normal skin. There existed a positive correlation between vessel count and the expression of HO-1 (r = 0. 761, P < 0.01) as well as the number of fibroblastic cells (r = 0. 731, P < 0.01) in the study groups.@*CONCLUSION@#HO-1 might play a important role in the angiogenesis of scar formation. The cause of these changes may be local. Over angiogenesis is one symbol of pathological scar.

Effect of Chinese drugs mixture on immune function of patients with extremely severe burn / 中国中西医结合杂志

<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>Chinese drugs mixture shows the regulatory effect on both cellular and humoral immune function in patients with ESB.</p>