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目的:观察五子衍宗丸含药血清对紫杉醇损伤的小肠隐窝细胞活力及凋亡蛋白Bcl-2、Bax、CytC、Caspase-3和Bcl-2/Bax表达的影响。方法:雄性SD大鼠随机分为五子衍宗丸组(五子衍宗丸6g·kg~(-1)·d~(-1)灌胃)和正常对照组(予等体积溶媒)。7d后两组制备血清。细胞培养分组为:正常对照组(正常大鼠血清5%、10%、20%)、模型对照组(正常对照组+紫杉醇)、实验组(紫杉醇+含五子衍宗丸大鼠血清5%、10%、20%)。MTS法测紫杉醇IC50浓度。MTS法检测五子衍宗丸不同浓度含药血清对紫杉醇(IC50浓度)干预后的IEC-6细胞活性。原位缺口末端检测法(TUNEL法)检测DNA断裂情况。蛋白质印迹法检测相关蛋白表达。结果:紫杉醇促进正常IEC-6细胞凋亡。五子衍宗丸对维持细胞形态、保持细胞活力有一定作用。TUNEL法显示实验组细胞凋亡明显少于模型对照组。与模型对照组比较,实验组较模型组Bax、Cytc、cl-Caspase-3显著降低(P<0.01),Bcl-2、Bcl-2/Bax显著升高(P<0.01)。结论:五子衍宗丸可通过调节Bcl-2/Bax信号通路减轻紫杉醇诱导的小肠隐窝细胞的凋亡损伤。
OBJECTIVE: To observe the effect of Wuzi Yanzong Pill serum on the intestinal crypt cell viability and the expression of Bcl-2, Bax, CytC, Caspase-3 and Bcl-2 / Bax in paclitaxel-injured small intestine. Methods: Male Sprague-Dawley rats were randomly divided into five sub-Yanzong pill group (intragastric administration of Wuzi Yanzong pill 6g · kg -1 · d -1) and normal control group (equal volume of vehicle). Serum was prepared in both groups after 7 days. The cell culture group was as follows: normal control group (normal rat serum 5%, 10%, 20%), model control group (normal control group + paclitaxel), experimental group (paclitaxel + Wuzi Yanzong Wan serum 5%, 10 %, 20%). MTS method to measure the concentration of paclitaxel IC50. MTS method was used to detect the IEC-6 cell activity of Wuzi Yanzong Pill with different concentrations of drug-containing serum after intervention with paclitaxel (IC50). In situ nick end detection (TUNEL method) to detect DNA fragmentation. Western blotting was used to detect related protein expression. Results: Paclitaxel promoted normal IEC-6 cell apoptosis. Wuzi Yanzong Pill to maintain cell morphology, maintain cell viability has a certain effect. TUNEL method showed that the experimental group apoptosis was significantly less than the model control group. Compared with model control group, Bax, Cytc and cl-Caspase-3 in experimental group were significantly decreased (P <0.01), Bcl-2 and Bcl-2 / Bax were significantly increased in experimental group compared with model group (P <0.01). Conclusion: Wuzi Yanzong pill can reduce paclitaxel-induced apoptotic injury of intestinal crypt cells through regulating Bcl-2 / Bax signaling pathway.