Quantitative evaluation of warm acupuncture-moxibustion in improvement of cartilage damage in rabbits with early knee osteoarthritis based on MR T2 mapping. / åŸºäºŽæ ¸ç£å ±æŒ¯T2 mappingå®šé‡è¯„ä»·æ¸©é’ˆç¸å¯¹æ—©æœŸå ”è†å ³èŠ‚éª¨å ³èŠ‚ç‚Žè½¯éª¨æŸä¼¤çš„ä¿®å¤ä½œç”¨.

OBJECTIVES: To observe the application value of MR T2 mapping for evaluating the effect of warm acupuncture-moxibustion on articular cartilage degeneration, and to observe the relationship between T2 value and expression of matrix metalloproteinases (MMP)-1 and MMP-13 of chondrocytes in rabbits with early knee osteoarthritis (KOA). METHODS: Thirty male New Zealand rabbits were randomly divided into blank control, KOA model and warm acupuncture-moxibustion groups, with 10 rabbits in each group. The early KOA model was established by right hind limb tubular plaster extension fixation method for 2 weeks. The rabbits of the warm acupuncture-moxibustion group received warm acupuncture-moxibustion stimulation at "Heding"(EX-LE2), "Neixiyan"(EX-LE4), "Waixiyan" (EX-LE5) and"Zusanli"(ST36) on the right hind limb for 15 min, once a day for 2 weeks. After intervention, MR T2 mapping of the right knee joint was performed in each group. The H.E. staining was used to evaluate the histopathological changes of cartilage, followed by giving a score according to the standards of Mankin scoring. The TUNEL method was used to analyze the apoptosis state of chondrocytes, and the positive expressions of MMP-1 and MMP-13 in the articular cartilage were detected by immunohistochemical staining. RESULTS: Compared with the blank control group, the Mankin score, chondrocyte apoptosis index, T2 value and the positive expressions of MMP-1 and MMP-13 in the cartilage tissue were significantly increased in the model group (P<0.01). Compared with the model group, the Mankin score, chondrocyte apoptosis index, T2 value and the positive expressions of MMP-1 and MMP-13 in the cartilage tissue were markedly decreased in the warm acupuncture-moxibustion group (P<0.01). The T2 value was positively correlated with the expression levels of MMP-1 and MMP-13 (P<0.01). H.E. staining showed disordered arrangement of chondrocytes and thinner cartilage layer in the model group, and a clear and relative ordered arrangement of chondrocyte in the warm acupuncture-moxibustion group. CONCLUSIONS: Warm acupuncture-moxibustion can reduce the T2 value of articular cartilage in early KOA rabbits, which is positively correlated with the decreased expression of MMP-1 and MMP-13 in the extracellular matrix of cartilage. The MR T2 mapping has certain value in evaluating the effect of warm acupuncture-moxibustion on KOA rabbits with early cartilage degeneration.

Warming moxibustion attenuates inflammation and cartilage degradation in experimental rabbit knee osteoarthriti.

OBJECTIVE: To explore the molecular mechanism of warming moxibustion (WM) in knee osteoarthritis (KOA). METHODS: The knee joints of 40 New Zealand rabbits were placed in a plaster cast in an extended position to establish a KOA model. The animals were randomly divided into four groups: the control group, model group, WM group, and diclofenac (DF) group. Hematoxylin-eosin staining and the modified Mankin score were applied to evaluate the histopathological changes. Chondrocyte apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Western blotting and real-time quantitative polymerase chain reaction were performed to measure the expression of interleukin-1α (IL-1ß), prostaglandin E receptor 3 (PTGER3), a disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-5 (ADAMTS-5), matrix metalloproteinase-13 (MMP-13), and C-terminal telopeptides of collagen type II (CTX-II) in cartilage tissues of the different groups. The concentrations of IL-1ß, PTGER3, and CTX-II in serum were detected by the enzyme-linked immunosorbent assay. RESULTS: Rabbits with KOA in the WM and DF groups showed significantly reduced cartilage erosion and Mankin scores, compared with the untreated rabbits. The number of TUNEL-positive cells observed in the WM group was much fewer than that in the model group. The expression of PTGER3, MMP-13, CTX-II IL-1ß, and ADAMTS-5 in cartilage tissues was remarkably downregulated following therapy with WM and DF. Moreover, a marked reduction was observed in the serum levels of IL-1ß, PTGER3, and CTX-II in the WM and DF groups. CONCLUSION: WM exerts favorable therapeutic effects on articular injuries of KOA by regulating the expression of inflammatory and cartilage degradation-related cytokines.

[Effect of warm needle moxibustion on morphological changes of cartilage and subchondral bone in knee osteoarthritis rabbits].

OBJECTIVE: To observe the effect of warm needle with moxibustion (WNM) on morphological changes of articular cartilage and subchondral bone tissues in knee osteoarthritis (KOA) rabbits, so as to explore whether WNM intervention can delay the development of KOA. METHODS: Forty male New Zealand rabbits were randomly divided into blank control, model, WNM and medication (Alendronate sodium) groups, with 10 rabbits in each group. A KOA model was established by immobilizing the right hind limb of rabbits with orthopedic casting tape for 6 weeks. The rabbits of the WNM group received WNM stimulation at "Neixiyan"(EX-LE4) "Waixiyan"(ST35) and "Heding"(EX-LE2) for 15 min, once a day for 4 weeks and those of the medication group received gavage of Alendronate sodium (150 µg·kg-1·d-1) once a day for 4 weeks. X-ray examination and magnetic resonance imaging (MRI) scanning were performed to observe the structure of the knee joints. Outcomes of X-ray examination were used to assess the degree of bone hyperplasia and joint space stenosis which were scaled according to Kellgren and Lawrence (K-L) standards, and those of MRI used to evaluate the degree of cartilage damage and bone marrow edema degrees which were respectively scored according to Recht scaling standards and semi-quantitative whole-organ MRI score (WORMS). Histopathological changes (degeneration degrees) of the articular cartilage were observed after H.E. staining, and given Mankin score. The ultrastructure of the cartilage surface and chondrocytes was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), respectively. RESULTS: Compared with the blank control group, the K-L grade, Recht grade, WORMS and Mankin score of the model group were significantly increased (P<0.05,P<0.01). After the intervention and in comparison with the model group, the K-L grade, Recht grade, WORMS and Mankin score of both WNM and medication groups were significantly decreased (P<0.05,P<0.01). Results of SEM showed severe defect, bulge and uneven surface of the cartilage with irregular growth and regeneration, and those of TEM showed degeneration and swelling of chondrocytes, margination of endonuclear chromatin, reduction in the number of organelles with disordered arrangement, extreme expansion of endoplasmic reticulum with detachment of the ribosome, unclear mitochondria, and disordered distribution of collagen fibers in the extracellular matrix in the model group, which was relatively milder in both the WNM and medication groups. Compared with the medication group, the Mankin score in the WNM group was evidently lower (P<0.05). CONCLUSION: WNM can effectively slow down the degeneration of cartilage and subchondral bone of KOA rabbits, delay the development of KOA.

Saccopetrins A and B, two novel gamma-lactones from Saccopetalum prolificum.

Two new gamma-lactones, saccopetrin A (1) and saccopetrin B (2), together with 10 known compounds have been isolated from the roots of Saccopetalum prolificum. Their structures were established by spectroscopic and chemical methods. The absolute configuration of 1 was determined by chemical transformation and the Mosher method. Cytotoxic activities were evaluated against several different cell lines. Compound 2 exhibited stronger cytotoxic activities than 1 against KB and HCT-8 cell lines at a concentration of 10(-5) mol/L.