Effects of Dachengqi Decoction on the Expression Levels of Bcl-2 and Bax Gene of Smooth Muscle Cells of Small Intestine in Rat with Multiple Organ Dysfunction Syndrome
|School||Dalian Medical University|
|Course||Traditional Chinese Medicine|
|Keywords||ultiple Organ Dysfunction Syndrome(MODS) Smooth Muscle Cells Apoptosis Bcl-2 Bax Dachengqi Decoction (DCQD)|
Background:Multiple organ dysfunction syndrome (MODS) is a clinical syndrome, which is induced by the primary disease, such as serious injury ( including shock, acute severe pancreatitis, et al), severe infections, extensive burns, surgical and obstetric major operations and oriented from systemic inflammatory response syndrome (SIRS). In MODS there are two or more organs dysfunction simultaneously or successively. It is the main cause of the death of patients with abdominal surgical diseases. The gastrointestinal tract is a reservoir of bacteria and considered to be the trigger and target site of SIRS and MODS. MODS may cause damage to gastrointestinal motility, bacterial translocation and impaired intestinal epithelial barrier function. The condition of gastrointestinal function is also thought to be the important criterion of evaluation the prognosis of critically ill patients. Promoting the recovery of gastrointestinal motility function could effectively prevent the patient from MODS deteriorating to multiple organ failure (MOF). The network of enteric nerve-ICC-smooth muscle is the basic unit of gastrointestinal motility. Cell apoptosis is the capacity of cells to undergo programmed cell death, which is controlled by a series of genes in the ontogeny course. Smooth muscle cells is the executor of the gastrointestinal motility, its normal structure and function has important significance for gastrointestinal motility. For smooth muscle cells, the original cancer albumen Bcl-2(B cell lymphoma/leukemia 2) protein family is important factors to the apoptosis, connecting and interacting with each other in the apoptosis to regulate the apoptosis, with the two opposite function, inhibition and facilitation. The two kinds of proteins are represented in the Bcl-2 and Bax(Bcl-2 associated X protein). Domestically, The Chinese Medicine are used in treatment of MODS leading to better therapeutic effect which displays distinctive predominance. Recent research has discovered that some Chinese Medicine can produce effect on several aspects of pathogenesis of MODS. Thus, it is important to prevention and cure of MODS that we find out more effective Chinese Medicine.Objective:To observe the distribution of Bcl-2 and Bax and the structure changes of smooth muscle cells in intestinal muscle of rats with MODS, and the effect of Dachengqi Decoction (DCQD) on Bcl-2 and Bax in intestinal muscle with MODS, to explore the mechanism of gastrointestinal motility disorders leaded by MODS, and to investigate the theory of effect of DCQD on MODS.Methods:One handred Wistar rats of both sexes weighing about 250g were randomly divided into three groups: control group(n=20), MODS group(n=40) and MODS+DCQD treated group(n=40). The model of MODS was established according to previous study. Briefly, one millilitre suspension of 8×10~8cfu/ml of Escherichia coli strain O127 H6, which contained 10%BaSO4, was injected under sterile conditions into the peritoneal cavity of the rats in the MODS and MODS+DCQD treated group. The rats of control group were injected 1ml of normal saline. The rats of DCQD treated group were administrated by gavage with DCQD (twice a day) before the suspension was injected. Twenty-four hours after injection, the proximal small intestine was taken.After some of the small intestine was put in 2.5% glutaraldehyde fixative, the tissue was cut into 1mm×2mm pieces, fixed, dehydrated, impregnated, embeded, semi-thin section was prepared, located on the light microscope, the thin slice was prepared. The structural changes of smooth muscle cells were observed, the images were collectted for statistical analysis.Other intestinal was for formaldehyde fixation, paraffin embedding, immunohistochemistry (SABC method) staining Bcl-2 and Bax, then coverslipping. The expression and distribution of Bcl-2 and Bax in the intestinal muscle were observed, The images were collected for statistical analysis.Results:Anatomical changes The gastrointestinal tracts were extremely dilated in the rats with MODS compared with in control group. Compared with the MODS group, the dilated gastrointestinal tracts was markedly alleviated in MODS+DCQD treated group.TEM observation Compared with the control group, the ultrastructural features of the smooth muscle cells in the MODS group displayed severe ultrastructural abnormalities. Injury was displayed in both the perinuclear cytoplasm and the smooth muscle cells processes, the nuclei of smooth muscle cells were shrunken, and the plasma membrane bleb was observed, smooth muscle cells appear apoptosis. Compared with the MODS group, the shape of ultrastructural features of smooth muscle cells in the DCQD group were better, the nuclei was nearly normal.Immunohistochemistry study Bcl-2 and Bax expression in cytoplasm of smooth muscle cells were observed in rats of various group, The characteristic of Bcl-2 and Bax immuno- reactivity was brown sediment.In control group, the brown sediment was evenly distributed in circular muscularis and longitudinal muscle layer. In MODS group, the expression of Bax was more than the control group (p<0.05), and the expression of Bcl-2 was not obvious different between the control group(p > 0.05). In the MODS+DCQD treated group, the expression of Bcl-2 was more than MODS group and control group(p<0.05), but the expression of Bax was less than the MODS group(p<0.05) and was not obvious different between the control group(p>0.05).Conclusion:There was extreme expansion of gastrointestinal and obvious signs of intestinal obstruction in MODS rat induced by bacterial peritonitis; DCQD can significantly alleviate the signs of paralytic intestinal obstruction.The structure of smooth muscle cells of small intestine in rat with MODS was damaged and smooth muscle cells appeared apoptosis. DCQD can protect structure of smooth muscle cells from destructing in rat with MODS.The expression of Bax was increased greatly in rat with MODS, DCQD could reduce the expression of Bax and increase the expression of Bcl-2 in rat with MODS. DCQD could protect gastrointestinal smooth muscle cells in MODS.