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目的:观察健脾化痰法对培养前脂肪细胞分化的影响,通过检测PPARγ2及PPARγ2 mRNA指标,探讨健脾化痰方的治疗作用机制。方法:以健脾化痰方、健脾方、化痰方、维甲酸及生理盐水给大鼠灌胃7天后,分离血清以培养并诱导前脂肪细胞分化,检测试验指标。结果:健脾化痰法可明显降低前脂肪细胞向脂肪细胞分化过程中PPARγ2 mRNA的表达,效果优于单纯健脾法或单纯化痰法治疗(P<0.05),时间和组别因素对PPARγ2 mRNA的表达影响较大;在第4天、10%浓度条件下,各组PPARγ2 mRNA表达量的变化趋势为:维甲酸组>化痰组>空白血清组>健脾组>健脾化痰组。Western Blot结果显示,健脾化痰组、健脾组、化痰组的PPARγ2表达量均明显低于维甲酸组和空白血清组,其中健脾化痰组表达量最小。结论:健脾化痰法可明显降低前脂肪细胞向脂肪细胞分化过程中PPARγ2及PPARγ2 mRNA的表达量,效果优于单纯健脾法或单纯化痰法。
Objective: To observe the effect of the method of invigorating the spleen and resolving phlegm on the differentiation of preadipocytes, and to explore the therapeutic mechanism of “Jianpi Huatan Fang” by detecting the mRNA of PPARγ2 and PPARγ2. Methods: The rats were gavaged with Jianpi Huatan Fang, Jianpi Fang, Huatan Fang, Retinoic acid and saline for 7 days. The serum was separated to induce differentiation of preadipocytes, and the test indexes were tested. Results: The method of invigorating the spleen and resolving phlegm could obviously decrease the expression of PPARγ2 mRNA during the differentiation of preadipocytes into adipocytes, which was better than that of the simple spleen and phlegm (P <0.05). The time and group factors had no significant effect on PPARγ2 The expression of PPARγ2 mRNA in each group tended to change on the 4th day under the condition of 10% concentration: Retinoid group> Phlegm group> Blank serum group> Jianpi group> Jianpi Huatan group . The results of Western Blot showed that the expression of PPARγ2 in Jianpi Huatan group, Jianpi group and Huatan group were significantly lower than that in retinoic acid group and blank serum group, and the expression of PPARγ2 in Jianpi Huatan group was the lowest. Conclusion: The method of invigorating the spleen and resolving phlegm can obviously reduce the expression of PPARγ2 and PPARγ2 mRNA during the differentiation of preadipocytes into adipocytes, which is better than the simple spleen - invigorating or simple phlegm - resolving method.