【摘要】 目的：体外实验探究银莱汤含药血清对胃肠积热合并肺炎RIN-14B细胞模型5-羟色胺（5-HT）合成、分泌的影响及机制；探讨银莱汤治疗胃肠积热合并肺炎的实验机制。方法：采用SD大鼠灌胃银莱汤制备含药血清；高热量饮食联合脂多糖（LPS）雾化制备模型组血清；0.9%氯化钠溶液灌胃制备正常组血清。CCK-8法检测不同血清对细胞活性的影响。采用模型组血清干预细胞复制胃肠积热合并肺炎体外模型，分为对照组、正常组、模型组、银莱汤组、TPH1抑制剂组、银莱汤+TPH1抑制剂组。ELISA法检测5-HT的表达；Western blot检测5-HT合成相关蛋白STAT5、pSTAT5(Tyr694)、TPH1及分泌相关蛋白TRPA1、TRPC4的表达。结果：与对照组比较，模型组5-HT水平显著降低（P<0.01）；与模型组比较，4%、8%银莱汤组5-HT水平显著升高（P<0.01）。与对照组比较，模型组STAT5、pSTAT5(Tyr694)、TPH1、TRPA1、TRPC4蛋白表达均显著降低（P<0.01）；与模型组比较，银莱汤组STAT5、pSTAT5(Tyr694)、TPH1、TRPC4蛋白表达显著升高（P<0.01,P<0.05）；与TPH1抑制剂组比较，抑制剂+银莱汤组TPH1、TRPA1、TRPC4蛋白表达显著升高（P<0.01,P<0.05）。结论：银莱汤可以提高胃肠积热合并肺炎细胞模型STAT5蛋白表达，并促进其磷酸化，进而提高TPH1的表达，促进5-HT合成。银莱汤可以促进TRPA1、TRPC4蛋白表达，促进5-HT分泌。更多还原

【Abstract】 Objective: To investigate the effects and mechanisms of Yinlai Decoction medicated serum on the production and secretion of 5-hydroxytryptamine(5-HT) in a RIN-14B cell model of gastrointestinal heat combined with pneumonia in vitro, and to explore the experimental mechanisms of Yinlai Decoction in treating this condition. Methods: SD rats were orally administered Yinlai Decoction to prepare medicated serum, while a high-calorie diet combined with LPS nebulization was used to prepare the model group serum, while normal saline was administered to prepare the control group serum. The CCK-8 assay was used to assess the effects of different sera on cell viability. The model group serum was used to intervene in cells to replicate an in vitro model of gastrointestinal heat accumulation combined with pneumonia. The cells were divided into the following groups: control group, normal group, model group, Yinlai Decoction group, TPH1 inhibitor group, and Yinlai Decoction+TPH1 inhibitor group. ELISA was used to measure 5-HT expression, while Western blot was used to detect the expression of 5-HT synthesisrelated proteins(STAT5, pSTAT5 [Tyr694], TPH1) and secretion-related proteins(TRPA1, TRPC4). Results: Compared with the control group, the 5-HT level in the model group was significantly decreased(P<0.01). In contrast, the 4% and 8% Yinlai Decoction group exhibited a significant increase in 5-HT levels compared with the model group(P<0.01). Compared with the normal group, the model group showed significantly reduced expression of STAT5, pSTAT5(Tyr694), TPH1, TRPA1, and TRPC4 proteins(P<0.01). Compared with the model group, the Yinlai Decoction group exhibited significantly increased expression of STAT5, pSTAT5(Tyr694), TPH1, and TRPC4 proteins(P<0.01, P<0.05). Compared with the TPH1 inhibitor group, the inhibitor+Yinlai Decoction group demonstrated significantly higher expression of TPH1, TRPA1, and TRPC4 proteins(P<0.01, P<0.05). Conclusion: Yinlai Decoction can enhance STAT5 protein expression and promote its phosphorylation in the cell model of gastrointestinal heat accumulation combined with pneumonia, thereby increasing TPH1 expression and facilitating 5-HT synthesis. Additionally, Yinlai Decoction can promote the expression of TRPA1 and TRPC4 proteins, thereby enhancing 5-HT secretion.更多还原