ABSTRACT
The present study investigated the therapeutic effect and mechanism of Di'ao Xinxuekang(DXXK) on non-alcoholic steatohepatitis(NASH) in mice. Sixty-five C57 BL/6 J mice were randomly divided into a normal group and an experimental group for model induction with the high-fat diet for 16 weeks. Then the mice in the experimental group were randomly divided into a model group, an atorvastatin group(4 mg·kg~(-1)·d~(-1)), and high-(200 mg·kg~(-1)·d~(-1)), medium-(60 mg·kg~(-1)·d~(-1)), and low-dose(20 mg·kg~(-1)·d~(-1)) DXXK groups, with 10 mice in each group. Drugs were administered by gavage for eight weeks. Serum lipid, liver lipid, serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione reductase(GSH-Px) were determined. Interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay(ELISA). The liver index was calculated. The liver pathological change and lipid accumulation were observed by HE and oil red O staining. The liver ultrastructure was observed by the transmission electron microscope. The mRNA and protein expression of nuclear factor-erythroid 2 related factor 2(Nrf2) and heme oxygenase-1(HO-1) was detected by real-time fluorescence-based quantitative PCR and Western blot, respectively. The results showed that compared with the normal group, the model group displayed serum lipid and liver lipid metabolism disorders, elevated transaminase, lipid deposition, steatosis, and inflammation, suggesting that the NASH model in mice was properly induced. Compared with the model group, the DXXK groups showed decreased serum lipid, liver lipid, ALT, AST, MDA, IL-1β, and TNF-α, increased SOD and GSH-Px, alleviated hepatic steatosis, ballooning, and inflammation, and up-regulated Nrf2 and HO-1 gene and protein expression. In conclusion, DXXK can significantly alleviate NASH in mice, which is related to the inhibition of oxidative stress and inflammatory damage by up-regulating the Nrf2/HO-1 signaling pathway.
Subject(s)
Animals , Mice , Drugs, Chinese Herbal , Inflammation/metabolism , Lipids , Liver , NF-E2-Related Factor 2/metabolism , Non-alcoholic Fatty Liver Disease/metabolism , Oxidative Stress , Signal Transduction , Superoxide Dismutase/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Objective:To investigate the effects of Di'ao Xinxuekang （DXXK） on NLRP3 inflammasome in mouse RAW264.7 macrophages and thoracic aorta of rats with atherosclerosis （AS）， so as to explore its anti-AS mechanism. Method:RAW264.7 cells were stimulated with oxidized low density lipoprotein （ox-LDL） and then intervened with MCC950 and DXXK. The contents of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） were determined by enzyme linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of Nod-like receptor protein 3 （NLRP3）， inflammasome adaptor protein apoptosis-associated speck-like protein containing CARD （ASC）， and cysteine-dependent aspartate-directed protease-1 （Caspase-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blotting. Sixty male SD rats were randomly divided into the normal group， model group， atorvastatin group （2.0 mg·kg<sup>-1</sup>）， as well as high-， medium-， and low-dose （100， 30， and 10 mg·kg<sup>-1</sup>） DXKK groups， with 10 rats in each group. The rats were exposed to the high-fat diet and vitamin D<sub>2</sub> for inducing AS. The blood lipid level was measured using an automatic biochemical analyzer， followed by the calculation of AS index （AI）. The contents of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> were determined by ELISA， and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in thoracic aorta were assayed by Real-time PCR and Western blotting. HE staining and Sirius red staining were conducted to observe the pathomorphological changes in the abdominal aorta and aortic sinus. Result:Compared with the normal group， the model group exhibited significantly increased TNF-<italic>α</italic> and IL-1<italic>β</italic> contents and up-regulated NLRP3， ASC， and Caspase-1 mRNA and protein expression in RAW264.7 cells （<italic>P</italic><0.01）. The above indexes in each drug administration group were significantly reduced in contrast to those in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. The comparison with the model group showed that cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and AI in each DXXK group significantly declined， while the high-density lipoprotein cholesterol （HDL-C） was significantly elevated （<italic>P</italic><0.05， <italic>P</italic><0.01）. The levels of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> and the mRNA and protein expression levels of NLRP3， ASC， and Caspase-1 in the thoracic aorta were decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Abdominal aortic lesions and fibrous hyperplasia of aortic sinus were significantly improved. Conclusion:DXXK has a significant anti-AS effect， which is possibly related to the inhibition of NLRP3 inflammasome.
ABSTRACT
Nonalcoholic fatty liver disease （NAFLD） is a metabolic stress-induced liver injury characterized by excessive lipid accumulation in hepatocytes， which is closely related to insulin resistance and genetic susceptibility. It falls into the category of "liver lump" in traditional Chinese medicine （TCM）. NAFLD affects about 25% of the population worldwide and has become a major burden of the world health care system. However， its exact pathogenesis remains unclear. Conducting the basic research on NAFLD is of great clinical significance and social value. As an important tool for NAFLD research， animal model plays a particularly important role in clarifying the pathophysiological mechanism of NAFLD. In recent years， the modeling methods for NAFLD in China and abroad have been constantly updated， and in particular， certain progress has been made in the duplication of TCM syndrome models. By consulting and sorting out the relevant literature published in recent years in China and abroad， the author summarized the replication methods of NAFLD animal models. This paper reviewed the advantages and disadvantages of models established via dietary induction （high-fat feed， high-fat and high-fructose feed， high-fat and high-cholesterol feed， and methionine choline-deficient feed）， models with genetic defects ［leptin-deficiency （Lepob/Lepob）， autosomal recessive diabetes gene homozygous deficiency （ob/ob）， Alms1 gene （foz/foz） mutation， and FATZO mice］ and exposure to special diets， and models for TCM syndromes （liver depression and spleen deficiency syndrome， phlegm-dampness syndrome， blood stasis syndrome， combined phlegm and stasis syndrome， and qi stagnation and blood stasis syndrome）， in order to provide reference for the preparation of more scientific， reasonable， economical， and convenient animal models of NAFLD， thus laying a foundation for in-depth study of the pathogenesis， prevention， and treatment of NAFLD.
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Objective::To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-κB (TLR/NF-κB) signaling pathway induced by monosodium urate in THP-1 cells, in order to explore the possible mechanism of anti-gout arthritis. Method::Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages, divided into normal group, model group, low, medium and high-concentration TSD groups (1, 3, 10 mg·L-1) and colchicine group (0.2 mg·L-1). Except the normal group, the other groups were stimulated with 400 mg·L-1 monosodium urate to replicate an inflammation model in vitro. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, the levels of inflammatory factors tumor necrosis factor-α(TNF-α ) and interleukin-1β(IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and NF-κB were detected by Western blot. The mRNA levels of TLR4, NF-κB and Pro-IL-1β were measured by real-time fluorescence quantitative PCR (Real-time PCR), and the nuclear shift of NF-κB p65 was detected by immunofluorescence. Result::0~32 mg·L-1 TSD has no effect on cell viability. Compared with the normal group, the secretion levels of inflammatory factors TNF-α and IL-1β in the model group were significantly increased (P<0.01), and the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were increased (P<0.01). Compared with the model group, 1-30 mg·L-1 TSD significantly down-regulated the secretion of inflammatory factors TNF-α and IL-1β (P<0.01), the expressions of key proteins (TLR4, MyD88 and NF-κB) and genes (TLR4, NF-κB and Pro-IL-1β) were decreased (P<0.05, P<0.01), and the NF-κB p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased, inhibiting the nuclear translocation of NF-κB p65. Conclusion::TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4, NF-κB and Pro-IL-1β mRNA and reducing the secretion of inflammatory factors TNF-α and IL-1β.
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<p><b>OBJECTIVE</b>To investigate the effect of total saponin of dioscorea (TSD) on uric acid excretion indicators in chronic hyperuricemia rats, and to study the correlation between blood levels of uric acid (SUA) and uric acid excretion indicators.</p><p><b>METHODS</b>Totally 90 SD male rats were randomly divided into 6 groups, i.e., the normal group, the model group, the benzbromarone group (10 mg/kg), the high, middle, and low dose TSD group (300,100, 30 mg/kg, respectively), 15 in each group. The chronic hyperuricemia model was prepared by potassium oxonate (200 mg/kg) and ethambutol (250 mg/kg) except in those of the normal group. All rats were intragastrically administered with corresponding medication once daily for 4 successive weeks since the third week. Contents of SUA and urine uric acid (UUA), serum creatinine (SCr), urine creatinine (UCr), blood urea nitrogen (BUN), and urine volume (UV) were measured on day 14 and day 28 respectively. Uric acid excretion indicators [including the amount of 24 h uric acid excretion (24 h UUA), fractional excretion of uric acid (FEUA), uric acid clearance (CUr), creatinine clearance (CCr), excretion of uric acid per volume of glomerular filtration (EurGF), and ratio of urinary uric acid to creatinine (UUA/UCr)] were calculated. The correlation between SUA and uric acid excretion indicators was analyzed by Pearson correlation analysis.</p><p><b>RESULTS</b>contents of SUA and SCr significantly increased, while the UUA, 24 h UUA, CCr, CUr, FEUA, EurGF, and UUA/UCr significantly decreased in the model group on day 14 and day 28. TSD could dose-dependently reduce the SUA level, significantly increase the UUA, 24 h UUA and CCr, Cur, FEUA, EurGF, showing an approximate effect to that of benzbromarone. But it had no effect on BUN, UCr, UUA/UCr, or UV of hyperuricemia rats. Correlation analysis showed that SUA level was positively correlated with SCr on day 14 and day 28 (r = 0.359, r = 0.306), but negatively correlated with CUr, FEUA, and CCr (r = -0.749 and -0.733; r = -0.669 and -0.646; r = -0.359 and -0.273). SUA level was not correlated with EurGF or UUA/UCr (r = 0.134 and 0.078; r = -0.057 and -0. 065). SUA level was negatively correlated with 24 h UUA on day 14 (r = -0.267), but not correlated with UUA or UCr on day 14. SUA level was negatively correlated with UUA and UCr on day 28 (r = -0.269, r = -0.275), but not correlated with 24 h UUA on day 28.</p><p><b>CONCLUSIONS</b>TSD could promote the excretion of uric acid and significantly increase the excretion of uric acid indicators. SUA was positively correlated with SCr, negatively correlated with Cur, FEUA, and CCr. FEUA and CUr were sensitive indicators of renal excretion of uric acid.</p>
Subject(s)
Animals , Male , Rats , Dioscorea , Chemistry , Hyperuricemia , Drug Therapy , Urine , Rats, Sprague-Dawley , Saponins , Pharmacology , Therapeutic Uses , Uric Acid , UrineABSTRACT
<p><b>OBJECTIVE</b>To study the preventive and therapeutic effects of total saponin of Dioscorea (TSD) on chronic hyperuricemia, and its effect on urate transporter 1 (URAT1) in rats.</p><p><b>METHOD</b>Ninety male rats were randomly divided into 6 groups: the normal group, the model group, TSD high-, medium- and low-dose (300, 100, 30 mg x kg(-1)) groups and the benzbromarone (10 mg x kg(-1)) group. Potassium oxonate and ethambutol were adopted to establish the chronic hyperuricemia model Since the third week, all the rats were intragastrically administered with drugs for 4 weeks, once a day, in order to determine their uric acid in serum and urine, uric acid excretion and xanthine oxidase (XOD). URAT1 mRNA and URAT1 protein expression in rat renal tubular cells were determined by RT-PCR and immunohistochemistry method respectively.</p><p><b>RESULT</b>Serum uric acid level of the model group increased significantly, while uric acid excretion decreased, with high expressions of renal URAT1 mRNA and URAT1 protein. TSD could dose-dependently reduce the serum uric acid level of chronic hyperuricemia rats, increase the concentration of uric acid and uric acid excretion in urine, and reduce renal URAT1 mRNA and URAT1 protein expression. Its effects were similar with that of benzbromarone, but with no significant effect on XOD and urinary volume of chronic hyperuricemia rats.</p><p><b>CONCLUSION</b>TSD has an obvious effect of anti-hyperuricemia It may reduce the reabsorption of uric acid by inhibiting the high expression of rat renal URAT1.</p>
Subject(s)
Animals , Male , Rats , Anion Transport Proteins , Genetics , Metabolism , Benzbromarone , Pharmacology , Dioscorea , Chemistry , Gout Suppressants , Chemistry , Pharmacology , Hyperuricemia , Blood , Drug Therapy , Genetics , Urine , Kidney Tubules , Metabolism , Rats, Sprague-Dawley , Saponins , Chemistry , Pharmacokinetics , Pharmacology , Uric Acid , Blood , Urine , Xanthine Oxidase , MetabolismABSTRACT
<p><b>AIM</b>To study the antidiabetic effects of cortex Moutan polysaccharide-2b (CMP-2b) in type 2 diabetes mellitus (T2DM) rats.</p><p><b>METHODS</b>The T2DM model rats were induced by a single intravenous injection of low dose streptozotocin (STZ) and intake of high sucrose-fat diet. CMP-2b was given to T2DM rats daily through gavage for 4-5 weeks. The body weight, water and food intake, fasting blood glucose (FBG), glucose tolerance, plasma lipids, serum insulin, and insulin receptor (Ins R) were determined.</p><p><b>RESULTS</b>Oral administration of CMP-2b significantly decreased water and food intake, FBG, total cholesterol (Tch), and triglyceride (TG), improved the impaired glucose tolerance (IGT), and remarkably raised the number of low affinity InsR and insulin sensitivity index (ISI) in T2DM rats.</p><p><b>CONCLUSION</b>CMP-2b may be useful for treating T2DM and its complications.</p>
Subject(s)
Animals , Male , Rats , Blood Glucose , Metabolism , Body Weight , Diabetes Mellitus, Experimental , Drug Therapy , Drugs, Chinese Herbal , Chemistry , Glucose Intolerance , Hypoglycemic Agents , Therapeutic Uses , Insulin , Blood , Liver , Metabolism , Paeonia , Chemistry , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Polysaccharides , Therapeutic Uses , Rats, Wistar , Receptor, Insulin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the therapeutic effect of Xinfeng Capsule (XFC) in treating adjuvant arthritis of rats and its effect on fas, fasL and bcl-2 expression in synovial membrane.</p><p><b>METHODS</b>Rats were randomly divided into the normal group, the model group, the model group, the methotrexate (MTX) group, the TPT group and the XFC group. Except for the rats in the normal group, animals were modelled to adjuvant arthritis with Freund's complete adjuvant, and the latter three groups were treated with MTX, Tripterygium wilfordii polysaccharide (TPT) and XFC respectively. The arthritis index (AI), change of body weight (BW) of rats were recorded, and the expressions of fas, fasL and bcl-2 in rats' synovial membrane were determined.</p><p><b>RESULTS</b>Compared with before treatment, the AI in the three treated groups was lowered significantly (P < 0.01, P < 0.05). The BW increment in the XFC group after treatment was insignificantly different to that in the normal group (P > 0.05), while it was significantly lower in the other two treated groups than that in the normal group and the XFC group (P < 0.01). Compared with the model group, fasL expression was increased in the XFC group significantly (P < 0.05), but bcl-2 expression decreased, fas expression showed insignificant difference (P > 0.05). Comparison of the three gene expressions between the three treated groups showed no significant difference (P > 0.05).</p><p><b>CONCLUSION</b>XFC could decrease AI in rats with adjuvant arthritis in the same way as MTX and TPT, it effected better in increasing BW than the latter two. The effect of XFC might be performed by its action in enhancing fasL expression, inhibiting bcl-2 expression and promoting apoptosis of proliferated synovial membranous cell so as to restrain hyperplasia of synovial membrane.</p>