目的:探讨加味参附汤对肾阳虚肺纤维化大鼠肺内细胞的影响及其相关的分子机制。方法:将60只雄性大鼠随机分为正常组、模型组和干预组,采用一次性博莱霉素A5气管插管给药加腺嘌呤灌胃28天复制肾阳虚肺纤维化大鼠模型。正常组大鼠注入等量生理盐水作为对照,模型组大鼠造模后生理盐水灌胃,干预组大鼠造模后加味参附汤灌胃,每组于实验第42天处死大鼠,留24h尿液检测17-羟皮质类固醇(17-OH—CS)含量,同时,流式细胞仪检测细胞的凋亡,Westernblot测定凋亡相关基因半胱氨酸天冬氨酸蛋白酶Caspase-3蛋白的表达。结果:干预组大鼠一般情况较模型组好,干预组大鼠细胞凋亡率低于模型组(P〈0.05),Caspase-3在干预组大鼠肺内的表达低于模型组(P〈0.05)。结论:加味参附汤可以改善大鼠肾阳虚症状,可能通过抑制Caspase-3蛋白的表达,从而缓解肺内细胞的凋亡减缓肺纤维化的发展。
Objective: To investigate the effect of modified Shenfu Decoction on lung cells of pulmonary fibrosis rats withkidney-yang deficiency and to explore its related molecular mechanism. Methods: Sixty male Wistar rats aged 6 weeks were randomly divided into normal group, model group and intervention group. Tracheal intubation of single-dose bleomycin A5 combined with intragastric infusion of adenine for 28 days was used to establish pulmonary fibrosis rat model of kidney-yang deficiency. Rats in the normal group were injected with isotonic saline. After modeling, the model group was given normal saline, intervention group was treated with modified Shenfu Decoction. On the experimental day 42, all of the rats were killed, 24-hour urinary 17- hydroxycorticosteroids(17-OH-CS) content was examined, flow cytometry was used to detect cell apoptosis, Western blot method was used to determime the expression of apoptosis-related gene aspartic proteinase Caspase-3. Results: The intervention group showed better general status, and had lower apoptotic rate and lower Capspase-3 expression level than the model group (P 〈 0.05). Conclusion: Modified Shenfu Decoction can relieve the symptoms of kidney-yang deficiency rats, and the mechanism is possibly through inhibiting the expression of Caspase-3 protein, which results into the alleviation of the lung cell apoptosis and the delay of pulmonary fibrosis development.