|
Into the fibroblasts (NRK49F) proliferation and activation of rat renal interstitial Objective: Chapa legislation calciferol induced by angiotensin Ⅱ (angiotensin Ⅱ, Ang Ⅱ), and to explore its mechanism. Methods: The the subculture NRK49F to added the following grouping various intervention factors: the paricalcitol final concentration of 10-6mol / L ,10-7mol / L ,10-8mol / L, Ang Ⅱ a final concentration of 10-6mol / L. 1 control group: NRK49F The culture medium without added interference factors; the ② Pa Li calciferol group: NRK49F culture solution were added to a final concentration of 10-6 mol / L ,10-7mol / L ,10-8mol, / L Pa established bone alcohols; ③ The Ang Ⅱ group: NRK49F culture medium added a final concentration of 10-6mol / L of Ang Ⅱ; ④ Ang Ⅱ Pa Li ossification alcohol group: NRK49F culture medium at the same time adding a final concentration 10-6mol / L of Ang Ⅱ and the final concentration of respectively 10 - 6mol / L ,10-7mol, L 10-8mol / L of Pago established calciferol. (1) using the WST-1 method detecting kidney Fibroblasts number was determined in the enzyme-linked immunosorbent detector at 450nm wavelength absorptiometry (OD) value, and the OD larger the value representative of the number of cells more. (2) cell immune chemical method (SP) was detected in each group cell α-SMA in protein expression, and mage-pro plus 6.0 analysis system α-muscle dynamic protein expression in a semi-quantitative analysis, to particle optical density IOD value reflect positive cells in total protein: the greater the IOD represent more positive cells in the total protein. (3) the flow cytometry NRK49F the cell cycle and apoptosis. All data were expressed as mean ± standard deviation ((?) ± s), using SPSS13.0 statistical software to analyze the results, the difference between the groups using single-factor analysis of variance design, pairwise comparisons using LSD method, P lt; 0.05 was statistically significant, P lt; 0.01 was significant. Results: 1 WST-1 assay kidney quality test results on the number of fibroblasts: 1 paricalcitol can inhibit the basic state NRK49F proliferation: OD value of the control group 0.3956 compared ,10-6mol / L, 10 - the 7mol / L ,10-8mol / L paricalcitol OD value 0.1288,0.2218,0.3208; the control group with paricalcitol group, pairwise comparisons between the paricalcitol group, the difference was statistically significance. ② paricalcitol can inhibit the proliferation of the Ang II-induced NRK49F: Compared with Ang Ⅱ group OD value 0.4853 ,10 mol / L ,10-7mol, / L ,10-8mol / L Pa Li calciferol Ang Ⅱ OD values decreased 0.1837,0.1466 and 0.1071; Ang Ⅱ group with paricalcitol group, pairwise comparisons between the paricalcitol group, the difference was statistically significant. 2α-SMA expression of test results: (1) paricalcitol can significantly inhibit α-SMA expression in the basis of the state NRK49F: 68.7438 compared with the control group IOD value ,10 mol / L of ,10-7mol / L, 10 Book mol / L paricalcitol IOD values ??were 0,0,35.6285; control group with paricalcitol group, pairwise comparisons between the paricalcitol group, the difference was statistically significant. The ② paricalcitol significantly induced by Ang Ⅱ the NRK-49Fα-SMA protein expression, thereby inhibiting the activation of the Ang II-induced NRK49F: ,10 mol / L ,10-7mol / L compared with the the Ang Ⅱ set of IOD value 237.54 10-8mol / L paricalcitol the Ang Ⅱ IOD value decreased 214.504,206.584 and 167,75.44; Ang Ⅱ group with paricalcitol group, pairwise comparisons between the paricalcitol group, the difference was statistically significance. 3 flow cytometry test results: in the basal state and after different concentrations of paricalcitol (10-6mol / L 107mo1 / L ,10-8mol, / L) 48 hours after induction, NRK-49F cells in G1 phase cells increased compared with the control group (22%, 16%, 13%), Ang Ⅱ group induced state, Parry calciferol intervention group G, phase cells than Ang Ⅱ group also significantly increased (22%, 13%, 1.1 %) (P lt; 0.05), especially in the 10-6mol / L concentration group; of 10-7mol / L ,10-8 mol / L concentration no significant difference, but that type of apoptosis paricalcitol None a significant effect (P gt; 0.05). Conclusion: (1) paricalcitol can inhibit the activation and proliferation of the basic state NRK49F the. (2) paricalcitol inhibit Ang Ⅱ can induce a NRK49F activation and proliferation. (3) the paricalcitol through so NRK49F stagnation reach the suppression NRK49F activation and proliferation in the G1 phase.
|