Study On The Effect Of Melittin On Il-1¦¢-Induced Expression Of Type Ii Collagen In Rat Lumbar Endplate Chondrocytes
Keywords
Melittin, Endplate Chondrocytes, Il-1¦¢, Type ¢ò Collagen, Rat
Abstract
Objective To observe the effect of Melittin on the expression of type II collagen in rat endplate chondrocytes (EPCs) induced by IL-1¦Â. Methods 4-week-old SD rats were cultured and morphologically observed, toluidine blue staining and immunofluorescence staining of type II collagen were used to culture EPCs in the lumbar spine. The third-generation EPCs were collected and the MTT method was used to determine the optimal concentrations of IL-1¦Â and Melittin for cell intervention. Then the third-generation EPCs were randomly divided into 4 groups: group A was the normal group without any drugs; group B was added with the optimal concentration of IL-1¦Â; group C was added with the optimal concentration of Melittin; group D was added with the optimal concentration of IL-1¦Â and Melittin; Western blot method was used to detect the expression of type ¢ò collagen after 48 hours of intervention. Results Observation under an inverted microscope showed that most of the cells in the first generation were polygonal. After the fifth generation, the cell proliferation ability gradually weakened and the cell morphology changed to spindle shape. Acidic mucoid substances (such as proteoglycans) in the cytoplasm are stained dark blue by toluidine blue. After immunofluorescence staining was used to label type II collagen, the cytoskeleton part was positively expressed (showing green fluorescence). The MTT assay showed that both IL-1¦Â and Melittin had an inhibitory effect on EPCs after 24 and 48 hours of intervention in a dose-dependent manner. It was finally determined that the optimal concentrations of IL-1¦Â and Melittin for intervention were 10 ng/mL and 10 ng/mL respectively. 1.0 ¦Ìg/mL. Western blot assay showed that compared with group A, the expression of type II collagen in group B was significantly reduced after IL-1¦Â intervention, and the expression of type II collagen was significantly increased in group C after Melittin intervention. However, there was no significant difference between group D and group A. The relative expression levels of type II collagen in groups A, B, C, and D were 0.991¡À0.024, 0.474¡À0.127, 1.913¡À0.350, and 1.159¡À0.297 respectively. Except for the difference between groups A and D, there was no statistical significance (P>0.05). , the differences between the remaining groups were statistically significant (P<0.05). Conclusion Research shows that Melittin has the effect of protecting endplate cartilage, providing relevant experimental basis for its use in the prevention and treatment of spinal degenerative diseases. For further information of this article and research, feel free to contact our team for asssitance. Original research was done by Hu Le, Zhang Changchun, Zhu Kun, Xu Gang, Han Zhongbing
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