肠吉泰对肠易激综合征内脏高敏感大鼠PAR2、PKCε、TRPV1蛋白表达的影响

Effects of Changjitai Decoction on the Expressions of PAR2,PKCεand TRPV1 Protein in Visceral Hypersensitivity Rats with Irritable Bowel Syndrome

目的:探讨肠吉泰对内脏高敏感模型大鼠的调节作用。方法:选择新生SD大鼠40只,按照随机数字表法随机分为空白组、模型对照组、阳性对照组及肠吉泰组,每组10只。参照Alchaer直肠醋酸刺激法建立IBS内脏高敏感大鼠模型。造模期间阳性对照组大鼠给予腹腔注射Capsazepine,其余各组除空白组外均给予相同体积的溶剂腹腔注射。从第8周开始,肠吉泰组每日给予0.423 g/mL的剂量中药灌胃,空白组、模型对照组和阳性对照组每日予以等量去离子水灌胃,各组均持续灌胃4周。干预4周后,采用结直肠气囊扩张法测定并记录大鼠腹壁反射(abdominal withdrawal reflex,AWR)评分,进行肠道敏感性评估;使用Western blot法检测大鼠结肠黏膜组织蛋白酶激活受体2(proteinase-activated receptors 2,PAR2)、下游物质蛋白激酶Cε(protein kinaseCε,PKCε)、瞬时感受器电位香草酸受体1(transient receptor potential vanilloid1,TRPV1)、磷酸化TRPV1(p-TRPV1)的表达。结果:当气囊压力为40、60 mm Hg(1 mm Hg≈0.133 kPa)时,模型对照组大鼠AWR评分较空白组明显升高(P<0.01);肠吉泰组和阳性对照组AWR评分与模型组比较均明显降低(P<0.05)。与模型对照组比较,阳性对照组及肠吉泰组PAR2、PKCε、TRPV1、p-TRPV1的表达显著降低(P<0.05)。结论:肠吉泰可能是通过下调PAR2、PKCε、TRPV1、p-TRPV1的表达,抑制其活化,从而改善IBS内脏高敏感。

Objective:To discuss the regulatory effect of Changjitai decoction on rat model with visceral hypersensitivity.Methods:Forty newborn SD rats were selected,divided into the blank group,model control group,positive control group and Changjitai decoction group according to random number table method,ten rats in each group,visceral hypersensitive rat model with IBS was established by referring to Alchaer rectal acetic acid stimulation.During the modeling period,rats in the positive control group were intraperitoneally injected with Capsazepine,and all other groups were intraperitoneally injected with the same volume of solvent except the blank group.From the 8th week,Changjitai decoction group was drenched with a dose of 0.423g/mL of herbs each day,the blank group,model control group and positive control group with the equivalent deionized water each day,different groups were medicated for four weeks consecutively by irrigation.After intervention for four weeks,colorectal balloon dilation was used to detect and record AWR scores of the rats,and to perform intestinal sensitivity;Western blot method was applied to detect the expressions of PAR2,PKCε,TRPV1 and p-TRPV1 in colonic mucosa of the rats.Results:When the balloon pressure was 40 and 60 mm Hg(1 mm Hg≈0.133 kPa),AWR scores of model control group increased notably compared with these of the blank group(P<0.01);AWR scores decreased obviously in Changjitai decoction group and positive control group compared with the model group(P<0.05).Compared with model control group,the expressions of PAR2,PKCε,TRPV1 and p-TRPV1 reduced notably in positive control group and Changjitai decoction group(P<0.05).Conclusion:Changjitai decoction could inhibit the activation of PAR2,PKCε,TRPV1 and p-TRPV1 possibly through down regulating their expressions,thus improving visceral hypersensitivity in IBS patients.