Abstract
Objective: To determine the astragaloside content in Qishen Huoxue Granule (QHG) by HPLC and to investigate the protective effect of astragaloside in septic rats. Methods: Dikma Diamonsil TM C 18 column was used. Mobile phase was methanol: acetonitrile (42:58) and detection was performed at 205 nm. Healthy rats were randomly divided into three groups, group A: sham operation (SO) group; group B: cecal ligation and puncture (CLP) group; and group C: CLP QHG treatment group. 72 h after model establishment, plasma tumor necrosis factor-α (TNF-α), plasma and peritoneal albumin concentrations, as well as mesenteric venular flow velocity in each group were measured and degree of intestinal mucosal injury was observed. Results: Average recovery was 100.7%, with a RSD of 2.20%. Sample concentration exhibited good linearity with peak area within a 0.01~0.2 mg/mL range. After CLP, plasma TNF-α concentration and albumin leakage in rats increased markedly; such increases were more significant in group B than group C (P<0.01). Changes in intestinal mucosal scores also presented the same trend (P<0.01). Mesenteric venular flow slowed down and plasma albumin concentration lowered; such decreases were more significant in group B than group C (P<0.05). Conclusion: QHG can reduce the inflammatory response and intestinal mucosal injury in rats with CLP-induced sepsis.
Highlights
Sepsis is a systemic inflammatory response syndrome (SIRS) caused by infectious factors
Astragaloside content is determined by HPLC and its sample concentration is found linear with peak area within a 0.01~0.2 mg/mL range
We found during the experiment that negative peak appeared under such mobile phase, which had poor peak shapes and were affected largely by the mobile phase, making it difficult to grasp the retention time
Summary
Sepsis is a systemic inflammatory response syndrome (SIRS) caused by infectious factors. Clinical features of sepsis are systemic high metabolic state, hyperdynamic circulatory state and excessive inflammatory response secondary to various severe infections; severe sepsis is complicated by one or more organ dysfunctions and accompanies varying degrees of coagulation disorders (WEYCKER et al, 2003; NICOLAI et al, 2015). From the perspective of traditional Chinese medicine, this study establishes a rat model of sepsis by CLP; determines and compares indices such as plasma TNF-α concentration, peritoneal albumin leakage, mesenteric venular flow velocity and intestinal mucosal injury score; observes the protective effect of QHG in septic rats; and explores its mechanisms of action. Astragaloside content is determined by HPLC and its sample concentration is found linear with peak area within a 0.01~0.2 mg/mL range. The method proposed is highly sensitive and reproducible, which can accurately determine astragaloside content in QHG. We aimed to determine the astragaloside content in Qishen
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