ABSTRACT
Osteoarthritis (OA), which is also called degenerative arthritis, is the leading cause of disabilities in the old people. The Chinese traditional herb Epimedium grandiflorum had long been found to attenuate osteoarthritis process, but the detailed mechanism was not clear. To study the mechanisms of E. grandiflorum in the treatment of osteoarthritis, rabbit osteoarthritis model combined with D-galactose was used. After different treatments for 10 weeks, cartilage sections were analyzed by immunohistochemistry for uPA, uPAR and PAI expression level. E. grandiflorum could significantly attenuate OA condition and decrease uPA, uPAR and PAI expression. The extract of E. grandiflorum, icariin also had a similar effect when compared with E. grandiflorum treatment alone. Rabbit chondrocytes were further isolated to be stimulated by TNFα combined with different reagents treatment. Here, icariin treatment significantly reduced nuclear factor kappa B NF-B (P65) activity, decreased uPA expression level and increased IBα protein level. The results indicated that E. grandiflorum and its extract icariin could attenuate OA condition, reduce the expression of uPA and uPAR and increase PAI in experimental rabbit model and this effect may be conducted by suppressing NF-kB activity by increasing IkBα level.
Subject(s)
Animals , Cartilage/metabolism , Chondrocytes/cytology , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Epimedium/metabolism , Female , Flavonoids/therapeutic use , Galactose/metabolism , I-kappa B Proteins/metabolism , Immunohistochemistry , Male , Medicine, Chinese Traditional , NF-kappa B/antagonists & inhibitors , NF-kappa B/metabolism , Osteoarthritis/drug therapy , Plasminogen Activator Inhibitor 1/metabolism , Rabbits , Receptors, Urokinase Plasminogen Activator/metabolism , Tumor Necrosis Factor-alpha/metabolism , Urokinase-Type Plasminogen Activator/metabolismABSTRACT
<p><b>OBJECTIVE</b>To study the molecular mechanisms of Fuyuan capsule serum containing, icariin and arasaponin R1 in the treatment of osteoarthritis from the urokinase-type plasminogen activator (uPA) system.</p><p><b>METHOD</b>Chondrocytes were isolated, cultured and identified using type II collagens immunostaining. After stimulating with TNF-alpha 10 microg x L(-1), 1 h, then the chondrocytes were treatment with glucosamine hydrochloride 25 g x L(-1), 20% Fuyuan capsule serum containing, icariin 12.5 mg x L(-1), arasaponin R1 125 mg x L(-1), icariin 12.5 mg x L(-1) + arasaponin R1 125 mg x L(-1). After 2 h, expression of uPA and nuclear factor kappa B (NF-kappaB P65) mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), the activities of NF-kappaB(P65) combine DNA were determined by electrophoretic mobility shift assays (EMSA), 1kappaBalpha were detected by Western blotting.</p><p><b>RESULT</b>Fuyuan capsule, icariin and arasaponin R1 could significantly reduce NF-kappaB (P65) activities and uPA mRNA expression, and increase expression of IkappaBalpha (P < 0.01), but no significant difference between all treatment groups.</p><p><b>CONCLUSION</b>Fuyuan capsule and its two main active ingredients, icariin and arasaponin R1, could protect chondrocytes from damage through reducing the NF-kappaB (P65) activities, increasing the express of IkappaBalpha and then reducing uPA of chondrocytes.</p>
Subject(s)
Animals , Male , Rabbits , Capsules , Cells, Cultured , Chondrocytes , Metabolism , DNA-Binding Proteins , Metabolism , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Flavonoids , Pharmacology , Therapeutic Uses , Gene Expression Regulation , NF-kappa B , Genetics , Metabolism , Osteoarthritis , Drug Therapy , Genetics , Metabolism , RNA, Messenger , Genetics , Urokinase-Type Plasminogen Activator , Genetics , MetabolismABSTRACT
Objective: To investigate effects of Erbie Decoction on the hepatocyte apoptosis-related factors Fas/Fas-L in rat with liver fi brosis,and to explore its anti-fi brosis mechanism. Methods: To Induce immune Liver fibrosis of rat model were established by intraperitoneal injection of pig serum. 48 rats were randomly divided into normal group, model group, pretreatment groups(high, middle and low dose groups of Erbie Decoction, and Fufang Biejia Ruangan Pills group. One week after the feeding adaptation, in addition to the normal group, other groups were given intraperitoneal injection of pig serum, each rat with 0.5ml, twice a week. At the same time, pretreatment groups were given intragastric administration daily. After 9 weeks, the expression of TGF-?1 and Fas/Fas-L were detected with immunohistochemistry assay. Results: Compared with normal group, in model group, TGF-?1 and Fas/Fas-L increased obviously(P
ABSTRACT
OBJECTIVE:To explore the effect of Fuyuan capsule on the mRNA expression of HIF-1? in kidney of sepsis rats. METHODS:Male SD rats were randomly divided into sham group,model group and Fuyuan capsule group (low dose and high dose) with 4 rats in each group. CLP operation was done in Fuyuan capsule groups after four days of intragastric administration. RT-PCR testing was applied to determined the expression of HIF-1a in 3 h,6 h,12 h and 24 h after model induction. The level of IL-6 was measured by ELISA method. Blood of heart was sampled for detection of renal function. HE staining was used to observe pathological change of kidney. RESULTS:As compared with sham group,the mRNA expression of HIF-1? increased gradually within 3 h and reached peak at 24 h(P
ABSTRACT
Objective To observe the effect of Fuyuan Capsule (FC) on vascular endothelial cell function of rats with qi deficiency and blood stasis.Method Forty-eight 16-month-old SD rats were equally randomized to 5 groups:FC group,positive control group (Qishen Capsule),model group and blank control group.Rat models of qi deficiency and blood stasis were induced by exertion,starvation and cold.The mice were given the drugs by gastric perfusion for 4 weeks.After treatment,endothelin (ET),6-keto-prostaglandin F_(1a) and nitric oxide (NO) levels were mea- sured.Results NO and 6-keto-PGF_(1a) levels were lower in the model group than those in the blank control group (P
ABSTRACT
Objective To explore the effect of medicated serum of Sangleng and Eshu on the expression of vascular endothelial growth factor and vascular endothelial cell proliferation induced by VEGF in vitro. Methods Medicated serum of Sangleng and Eshu was used to culture human umbilical vein endothelial cell (HUVEC-1) induced by VEGF. The morphologic changes of HUVEC-1 were observed with phase contrast microscope, and cell proliferation was detected by MTT method, and the expression of vascular endothelial growth factor protein and mRNA in endothelial cells was detected by Western blotting and RT-PCR. Results The medicated serum of 5.0, 2.5 g?kg~ -1 ?d~ -1 Sangleng and Eshu could cause arrangement disorder in the normal umbilical vein endothelial cells. The medicated serum of 5.0 g?kg~ -1 ?d~ -1 Sangleng and Eshu (10%, 5%, 2.5%) and medicated serum of 2.5 g?kg~ -1 ?d~ -1 (10%) could inhibit vascular endothelial cell proliferation remarkably (P