1.Effects of anthraquinones from Cassia obtusifolia L. on cholesterol biosynthesis in cells
Chuhua LI ; Xue LI ; Kunyang FANG ; Baoliang GUO
Chinese Journal of Tissue Engineering Research 2008;12(33):6593-6596
BACKGROUND: The effective chemical constituents and the precise action mechanism of Cassia obtusifolia L. seed to rats with hypercholesterolemia are not clear. OBJECTIVE: To investigate effects of anthraquinones from Cassia obtusifolia L. on hypolipidemic activity and endogenous cholesterol biosynthesis in rats with experimental hyperlipoidemia, and to explore the effective compound and the mechanism of Cassia obtusifolia L. seed on hypolipidemic action. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed from September 2003 to May 2004. All rats were raised and tested at the College of Life Sciences, South China Normal University. Blood sample was collected from the tail vein at the Institute of Biotechnology to detect blood lipid, survival rate, and to culture cells. MATERIALS: Forty-five male Sprague Dawley (SD) rats were used to establish animal models of hypedipoidemia by intragastrically with fat emulsion. Cassia obtusifolia L. seed was purchased from Guangzhou Dispensary, China, and further identified by South China Plant Institute. Anthraquinones were extracted from Cassia obtusifolia L. by the Institute of Biotechnology of South China Normal University. METHODS: Forty-five male SD rats were randomly divided into three groups (n=15): a control group, 80 mg/kg and 20 mg/kg anthraquinones groups. Rat models in each group were given fat emulsion in the morning and afternoon 2 days after model induction, once in morning. Rat modes were treated with corresponding doses of drugs in the two experimental groups. Rat models were administrated with the same volume of saline in the control group, once a day, for 20 days. MAIN OUTCOME MEASURES: Effect of anthraquinones on endogenous cholesterin in Chinese hamster oocytes was measured by amphotericin B cell models. The levels of serum total cholesterol, triglyceride, low-density iipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) were detected by enzyme endpoint method. Survival rate of Chinese hamster oocytes (A570) was tested by methyl thiazolyi tetrazolium (MTT) with spectrophotometric determination. RESULTS: Anthraquinones significantly reduced total cholesterol, triglyceride, LDL-C levels and increased HDL-C levels in hyperlipidemic rats in a dose-dependant pattern. Anthraquinones elevated the survival rate of Chinese hamster oocytes. CONCLUSION: Anthraquinones can decrease blood lipid levels. Inhibition of cholesterol synthesis of anthraquinones may be one of the underlying mechanism involved in decreasing blood lipid.
2.Effect and Mechanism of Action of Epimedii Folium Polysaccharides on Mice with Exercise-induced Fatigue Based on p38 MAPK/NF-κB Signaling Pathway
Yue ZOU ; Min XIAO ; Yuhao MENG ; Kunyang TANG ; Xiaocui JIANG ; Zhipeng FANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):20-28
ObjectiveTo study the effects of Epimedii Folium polysaccharides on mice with exercise-induced fatigue and explore its possible mechanism of action. MethodICR male mice screened by swimming training were randomly divided into a control group, model group, vitamin C group, and low, medium, and high dose groups of Epimedii Folium polysaccharides, with eight mice in each group. The exercise-induced fatigue model was established by weight-bearing swimming training in each group except for the control group. After two weeks of weight-bearing swimming, the Epimedii Folium polysaccharide groups were given 100, 200, 400 mg∙kg-1 of Epimedii Folium polysaccharides by gavage, and the vitamin C group was given 200 mg∙kg-1 of vitamin C by gavage. The control group and the model group were given equal amounts of saline for 14 d. At the end of the experimental period, the body mass of the mice in each group and the time of last swimming due to exhaustion were recorded. Serum urea nitrogen (BUN), lactic acid (LA), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidation (GSH-Px), myoglycogen (MG) in skeletal muscle, hepatic glycogen (HG) in the liver were detected by kits. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in muscle tissue. Western blot was used to detect the protein expression of p38 mitogen-activated protein kinase (p38 MAPK), phosphorylation (p)-p38 MAPK, extracellular signal-regulated kinase1/2 (ERK1/2), nuclear factor-κB (NF-κB), p-NF-κB, interleukin-1β (IL-1β), and interleukin-6 (IL-6) in muscle tissue. The immunofluorescence (IF) method was used to detect the expression of tumor necrosis factor-α (TNF-α) in skeletal muscle tissue of mice in each group. ResultCompared with the control group, the body mass of mice in the model group decreased, and the time of last swimming due to exhaustion decreased (P<0.01). In addition, there were significantly higher serum levels of the fatigue metabolites LA, LDH, BUN, and lipid peroxidation product MDA (P<0.01) and decreased levels of MG, HG, SOD, and GSH-Px (P<0.01). The protein expressions of p-p38 MAPK, ERK1/2, p-NF-κB, IL-1β, IL-6, and TNF-α in skeletal muscle tissue were significantly higher than those of the control group (P<0.01). Compared with the model group, the body mass and time of last swimming due to exhaustion of the mice in the low, medium, and high dose groups of Epimedii Folium polysaccharides and the vitamin C group were increased (P<0.05, P<0.01), and the contents of LA, LDH, BUN, and MDA were significantly decreased (P<0.05, P<0.01). The levels of MG, HG, SOD, and GSH-Px increased (P<0.05, P<0.01), and the protein expression levels of p-p38 MAPK, ERK, p-NF-κB, IL-1β, IL-6, and TNF-α in skeletal muscle tissue decreased (P<0.05, P<0.01). ConclusionEpimedii Folium polysaccharides can play a role in alleviating exercise-induced fatigue by inhibiting the p38 MARK/NF-κB signaling pathway, thereby reducing the accumulation of metabolites, improving the activity of antioxidant enzymes, increasing the glycogen content of the body, and reducing inflammation in skeletal muscle.