| ObjectivePopulation aging has become a global public health problem. With the increasing elderly population, elderly patients with liver and kidney disease also increases. Liver and kidney are vital organs in humans which have significant morphological and functional changes associated with aging. It has a very important significance for the public health development of aging society to study the pathogenesis of liver and kidney damage of aging, explore the effective drugs for prevention and treatment of liver and kidney damage of aging, provide basic research data for clinical treatment. This paper aims to explore the ameliorative effect of CO2 supercritical fluid extract of Ligusticum Chuanxiong Hort (CX) and Angelicae Sinensis Radix (DG) against D-galactose (D-gal) induced injury in the mouse liver and kidney.MethodsFirstly, the gas chromatography-mass spectrometry technology(GC-MS) was selected to analyzed the chemical composition of CO2 supercritical fluid extract of Ligusticum Chuanxiong Hort and Angelicae Sinensis Radix, and the determination of their in vitro antioxidant activity was accomplished.Our research selected Kunming male mice (SPF grade, eight weeks old), subcutaneous injection of D-gal 200 mg/(kg) on the back to induce the model of liver and kidney damage in aging mice. By comparing the weight of mice and organs index, determination of liver and kidney function index, observation of the histologic changes, we evaluated the ameliorative effect of CO2 supercritical fluid extract of Ligusticum Chuanxiong Hort and Angelicae Sinensis Radix against D-galactose induced injury in the mouse liver and kidney. And we attempted to discuss its mechanism through antioxidant and anti-inflammatory.1 The evaluation of the ameliorative effect CX and DG against D-galactose Induced Injury in the Mouse Liver and KidneyMice were randomly divided into eight groups,10 mice each group, namely the normal group, D-galactose model group (D-gal), the positive control vitamin E group (80 mg/kg) (VE), CO2 Supercritical Fluid Extract of Ligusticum Chuanxiong Hort (50,100 mg/kg) (CX), CO2 Supercritical Fluid Extract of Angelicae Sinensis Radix three dose groups20,40,80 mg/kg (DG). Subcutaneous injection of D-ga1200mg/(kg · d) mice in each group, at dose of 0.1ml/10g bodyweight, normal group receive saline,0.1 ml/10g body weight. Gavage at the same time modeling, the control group and D-gal model group were given an equal volume of saline, both once daily for 8 weeks. The weight of the mice was record once a week.24 hours after the last time administrate the drug, take the mice orbital blood, measure the liver function index (ALT, AST) and renal function index (BUN, CRE) according to the kit method. Mice were sacrificed and then dissected to get the thymus, spleen, liver and kidneys, weighed and recorded and viscera index, liver and kidney paraffin-embedded sections with HE staining were made to observe the pathological changes.2 The mechanism of the ameliorative effect of CX and DG against D-galactose induced injury in the mouse liver and kidney.The lipid peroxide malondialdehyde (MDA) in liver and kidney was measure by using biochemical quantitative analysis as well as the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px). The RN A expression of these three antioxidant enzyme was analyzed by real-time quantitative detection of nucleic acid amplification. In vitro anti-oxidation test was carried out to verificate the antioxidant capacity of CX and DG. The protein expression of iNOS, COX2, NF-κB was analyzed by using western blot.Results1 The chemical composition of CX and DG analyzed by GC-MSThe main chemical compositions of CX and DG is ligustilide, the content of ligustilide in DG was higher than that in CX, respectively,84.18% and 59.221%. As major pharmacologically active ingredient of both CX and DG, the content of ligustilide may affect the strength of their pharmacological effects.2 The determination of CX and DG in vitro antioxidant activityBoth CX and DG has a certain antioxidant activity in vitro, and the antioxidant activity of DG is slightly higher than CX.3 The evaluation of the ameliorative effect CX and DG against D-galactose Induced Injury in the Mouse Liver and KidneyThe data showed that the body weight and organ index of model group is lower than those of the normal group. Both CX and DG increased the weight and organ index of mice, made it close to normal, but did not reflect the concentration dependent manner. CX and DG reduced the liver and kidney function index, and reflected the concentration dependent manner. Pathological histology observation found that the model groups appear damage in the liver and kidney, CX and DG ameliorative the damage of the tissue of liver and kidney.4 The mechanism of the ameliorative effect of CX and DG against D-galactose induced injury in the mouse liver and kidney.In vitro antioxidant experimental results indicate that CX and DG has a certain antioxidant capacity; CX and DG significantly decreased the lipid peroxide malondialdehyde (MDA) level in mice liver and kidney; and raise the activity of SOD, CAT and GSH-Px; Real-time fluorescent quantitative nucleic acid amplification testing results showed that the CX and DG increased the RNA expression of SOD, CAT and GSH-Px; Western blot result showed that CX and DG significantly lower the expression of iNOS, COX2 and NF-K B protein.ConclusionThe main chemical compositions of CX and DG is ligustilide.They both ameliorated the injury in the mouse liver and kidney induced by D-galactose, its potential mechanism may be related to the improvement of inflammation and the regulation of oxidative stress. |
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