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Pathological mesangial cell proliferation and an increase in mesangial matrix is ??the most common glomerulonephritis pathological changes, most with chronic progressive glomerulonephritis development trend in the progression of mesangial cells not only are the victims, And through the secretion of inflammatory cytokines and extracellular matrix actively participate in the development of glomerular inflammation and promote glomerulosclerosis. Including transforming growth factor β1 (TGF-β1) can stimulate the secretion of extracellular matrix, is an important factor to promote fibrosis, one of its activation can stimulate mesangial cells to synthesize Ⅱ, Ⅳ collagen, FN and other important component of the extracellular matrix element, increasing the mesangial area mesangial extracellular matrix accumulation, accelerate the progress of glomerular sclerosis. Apoptosis and proliferation in glomerular lesion formation and dissipation process also plays a very important role. Recent studies show that in some self-limiting proliferative glomerular diseases, the proliferation of mesangial cells through apoptosis mechanism can relieve itself, the glomerular structure back to normal. For the treatment of proliferative nephritis, now the preferred drugs are corticosteroids, however, a considerable part of its dependence patients, resistance, or even serious side effects, followed by cyclophosphamide, chlorambucil, cyclosporin vitamin A and other immunosuppressive therapy came into being, but there are still some patients difficult to obtain satisfactory results, the development of new drugs will become a research hotspot. Derived from traditional Chinese medicine monomer Bufalin toad Bufo bufo and boxed ear glands and skin gland serous, molecular formula C24 H34 O4, relative molecular mass of 386.5, research shows that this ingredient has anti-cancer, cardiac, toxicity and anesthesia and other effects, it may be inhibited by interfering with the cell cycle and proliferation of leukemia cells by changing the apoptosis-related genes such as bcl-2 family, abnormal activation of MAPK pathways induced by a variety of tumor cell differentiation and apoptosis. We speculate Bufalin on mesangial cells may have a similar effect, but on Bufalin glomerular mesangial cells poorly understood, therefore, this study observed the one hand, the GMC Bufalin abnormal proliferation and extracellular matrix secretion, on the other hand, observed abnormal proliferation Bufalin GMC apoptosis, is to be developed for clinical treatment of proliferative glomerulonephritis provide a theoretical basis. The first part Bufalin rat mesangial cell proliferation and extracellular matrix secretion Objective: To investigate Bufalin (bufalin) on lipopolysaccharide (LPS)-induced rat glomerular mesangial cells (GMC) proliferation and Extracellular matrix (ECM) secretion. Methods: Cultured rat mesangial cells were divided into control group, LPS stimulation group (LPS 5mg / L) and Bufalin intervention group (LPS 5mg / L and Bufalin 1 × 10-8mol / L), the application sets trypan blue exclusion staining assay Bufalin the GMC cytotoxicity by MTT, flow cytometry GMC proliferation changes, RT-PCR detection of interleukin -6 (IL-6) and transforming growth factor-β1 (TGF- β1) of mRNA expression, ELISA assay GMC supernatant fibronectin (FN) and TGF-β1 expression. Results: MTT results showed Bufalin concentration 1 × 10-8mol / L and above GMC proliferation in a dose-dependent inhibition (P lt; 0.01). Cell cycle analysis showed Bufalin percentage of S phase cells decreased compared with LPS group (P lt; 0.01). Bufalin intervention group compared with LPS IL-6 and TGF-β1 mRNA expression is decreased (P lt; 0.01), FN, TGF-β1 protein secretion decreased (P lt; 0.01). Conclusion: Bufalin right after LPS mesangial cell proliferation and extracellular matrix secretion was inhibited. The second part Bufalin on LPS-induced mesangial cell apoptosis Objective: To investigate Bufalin (bufalin) on LPS (lipopolysaccharides, LPS)-induced rat mesangial cells (glomcrular mesa ngial cell, GMC) apoptosis. Methods: Cultured rat mesangial cells were divided into control group, LPS stimulation group and intervention group Bufalin, application trypan blue exclusion staining Bufalin cytotoxicity against GMC, application TEM mesangial ultrastructural alterations, using RT-PCR detection of Bax and bcl-2 mRNA expression, Western blot assay of Bax and bcl-2 protein expression. Results: The intervention group TEM Bufalin early apoptotic morphological changes. RT-PCR and Western blot showed Bufalin group than in the LPS group increased expression of Bax, bcl-2 expression was decreased, the differences were statistically significant (P lt; 0.01), after the intervention Bufalin, Bax excessive compared with bcl-2 expression . Conclusion: Bufalin may regulate Bax and bcl-2 expression and the relative after lipopolysaccharide induced mesangial cell apoptosis.
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