舒胃汤对功能性消化不良大鼠小肠ICC细胞IP3R，RyR表达的影响

目的:通过观察舒胃汤对功能性消化不良(functional dyspepsia,FD)模型大鼠小肠Cajal间质细胞(interstitial cells of Cajal,ICC)中IP3受体(inositol 1,4,5-triphosphate receptor,IP3R),Ry受体(ryanodine receptor,RyR)蛋白及mRNA表达的影响,从钙离子通道调节角度探讨舒胃汤对FD的治疗作用及机制。方法:取Wistar大鼠15只,随机分成空白组、模型组与舒胃汤组(30.68 g·kg~(-1))。模型组与舒胃汤组按复合病因造模法处理21 d复制FD模型,灌胃给药14 d后取血制备含药血清。取2~3周Wistar乳鼠小肠平滑肌原代ICC细胞,传代培养后分为空白血清组(A组),模型血清组(B组),舒胃汤5%含药血清组(C组),舒胃汤10%含药血清组(D组),舒胃汤15%含药血清组(E组)。待细胞融合至60%~80%时分别加入对应组别血清,继续孵育24 h。提取细胞蛋白及总RNA,蛋白免疫印迹法(Western blot)检测IP3R,RyR蛋白表达,实时荧光定量PCR法(Real-time PCR)检测IP3R,RyR mRNA表达。结果:Western blot检测发现,与A组比较,B组IP3R-1,IP3R-2,IP3R-3,RyR-1,RyR-2,RyR-3蛋白表达均明显降低(P0.01);与B组比较,D,E组IP3R,RyR蛋白表达明显升高(P0.05,P0.01)。Real-time PCR检测发现,与A组比较,IP3R,RyR mRNA表达均明显降低(P0.01);与B组比较,D,E组IP3R,RyR mRNA表达明显升高(P0.01)。结论:舒胃汤可通过上调FD模型大鼠小肠ICC细胞内IP3R,RyR蛋白及mRNA表达,发挥对功能性消化不良疾病的治疗作用。

Effect of Shuwei Decoction on Expression of IP3R and RyR in Gastric Antrum ICC Cells in Rats Model of Functional Dyspepsia

Objective:To investigate the effect of Shuwei decoction on expressions of IP3 receptor (inositol 1,4,5-triphosphate receptor, IP3R), ryanodine receptor(RyR)protein and mRNA expression of intestinal cells of Cajal (ICC) of rats model of functional dyspepsia (FD), in order to explore the mechanism of Shuwei decoction in the treatment of FD in the aspect of calcium channel adjustment. Method:Totally 15 Wistar rats were randomly divided into the blank group, the model group and the Shuwei decoction group (30.68 g·kg^-1). The model group and the Shuwei decoction group were treated by the compound etiological modeling method for 21 day to replicate the FD model. After intragastric administration for 14 d, blood was collected to prepare drug-containing serum. Intestinal smooth muscle in subcultured ICC cells was taken from 2-3-week-old Wistar rats, and were divided into blank serum group (group A), model serum group (group B), Shuwei decoction 5%-containing serum group (group C), Shuwei decoction 10%-containing serum group (group D), and Shuwei decoction 15%-containing serum group (group E). When the cells were fused to 60%-80%, corresponding serums were added to each group and incubated for 24 h. IP3R and RyR were detected by Western blot, and IP3R and RyR mRNA expressions were detected by Real-time PCR. Result:Western blot results showed that compared with group A, the expressions of IP3R-1, IP3R-2, IP3R-3, RyR-1, RyR-2, RyR-3B protein in group B were significantly decreased (P<0.01). Compared with group B, the expressions of IP3R, RyR protein in group D and E were significantly increased (P<0.05, P<0.01). Real-time PCR results showed that mRNA expressions of IP3R and RyR were significantly decreased compared with group A (P<0.01). Compared with group B, mRNA expressions of IP3R and RyR in group D and E were significantly increased (P<0.01). Conclusion:Shuwei decoction has a therapeutic effect on functional dyspepsia by up-regulating IP3R, RyR and mRNA expression.