ABSTRACT
BACKGROUND: Mechanical load is crucial for the degeneration of chondrocytes and the development of osteoarthritis. Clematis chinensis can improve the inflammatory microenvironment of osteoarthritis, but its effect on mechanical load-induced degeneration of chondrocytes has not been elucidated. OBJECTIVE: To study the effect and mechanism of the extracts of Clematis chinensis on the degenerative changes of chondrocytes induced by intermittent cyclic mechanical tension (ICMT) in vitro. METHODS: Chondrocytes of the rabbit knee joint were isolated by type II collagenase digestion method and identified by Alcian blue staining. There were five groups in the experiment: Blank group, ICMT group, high-, medium- and low-dose Clematis chinensis groups. There was no intervention in the blank group, and the other groups were subjected to ICMT (10% tensile strength, 0.5 Hz, 8 hours per day, for a total of 2 days) for inducing chondrocyte degeneration. Three Clematis chinensis groups were concurrently given 0.5, 1, 2 g/L extracts of Clematis chinensis, respectively. The intervention time was 48 hours. Cell counting kit-8 assay was used for detection of chondrocyte proliferation. FITC-phalloidin staining was used for observation of cytoskeleton morphology. Real-time quantitative PCR and western blot assay were used for determination of collagen type II, matrix metalloproteinase 13, and transforming growth factor β at protein and gene levels, respectively. RESULTS AND CONCLUSION: (1) Compared with the blank group, the cytoskeleton of chondrocytes stimulated by ICMT was long- stretched, the proliferation activity of chondrocytes decreased, and the expressions of collagen type II and transforming growth factor β were down-regulated, while the expression of matrix metalloproteinase 13 was up-regulated. (2) Compared with the ICMT group, the extract of Clematis chinensis could promote the proliferation of chondrocytes, up-regulate the expressions of transforming growth factor β and collagen II, and down-regulate the expression of matrix metalloproteinase 13 in a concentration-dependent manner. To conclude, the extract of Clematis chinensis can inhibit the catabolism of chondrocyte induced by ICMT through regulating the expression of transforming growth factor β, promote the synthesis of extracellular matrix of chondrocytes, and maintain the phenotypic stability of chondrocytes.
ABSTRACT
This study was aimed to optimize extraction process of formula granules of Clematis chinensis Osbeck and establish its qualitative discrimination method. With oleanolic acid, hederagenin and extract rate as indexes, extraction technology conditions of formula granules of C. chinensis Osbeck were investigated by L9 (34) orthogonal test method. TLC was used in the qualitative discrimination of formula granules of C. chinensis Osbeck. The results showed that optimum extraction technology was: adding 12 times of water; soaking for 0.5 h; cooking for 1 h; and extracting for 3 times. TLC had the diagnostic characteristic for the distinct spot which illustrated the specificity. It was concluded that the extraction process was reasonable, simple and feasible with high extraction efficiency of ef-fective contents, which can be the scientific basis for production and quality control of formula granules of C. chin-ensis Osbeck.
ABSTRACT
Objective: To study the antioxidant activity of Clematis chinensis Osbeck polysaccharide(CCP).Methods:① To assay the scavenging activity of CCP on hydroxyl radical and superoxide anion in vitro.② To analyze the effect of CCP on red blood cell autoxidation hemolysis induced by H2O2 by colorimetry.③ To study the effect of CCP on acute hepatic injury of the mice induced by carbon tetrachloride.Results: CCP could eliminate the hydroxyl free radical and the superoxide anion free radical,reduce red blood cell autoxidation hemolysis induced by H2O2,raise the activities of SOD and GSH-Px in hepatic injury mice's serum and liver apparently,reduce the content of MDA and liver index notably.Conclusion: CCP has the significantly action of anti oxidation activity in vitro and in vivo which relates to removing oxygen free radical.