乌梅丸及拆方对溃疡性结肠炎模型大鼠炎症因子和神经肽表达的影响

To explore the effect on Fructus Mume Pills and its compatibility against ulcerative colitis model rats via inflammation cytokines and neuropeptides

目的观察乌梅丸对溃疡性结肠炎大鼠的作用机制及配伍特点。方法83只SD雄性大鼠随机分为8组,其中空白对照组、全方组、酸味组、苦味组、辛味组、甘味组、阳性药组(给予柳氮磺胺吡啶治疗)各10只,模型组13只。采用4%乙酸灌肠法诱导溃疡性结肠炎大鼠模型。造模24小时后给药,全方组大鼠每日给药量为900 mg/kg,酸味组270 mg/kg,苦味组235 mg/kg,辛味组340 mg/kg,甘味组110 mg/kg,阳性药组300 mg/kg,以10 mL/kg大鼠体质量的容量灌胃给药,每日2次,连续7天。模型组和空白对照组灌以等量生理盐水。通过疾病损伤指数(disease damage inde,DAI)对疾病模型进行评分,黏膜病理损伤通过结苏木素—伊红染色观察。采用ELISA法检测血清炎症细胞因子白介素(interleukin,IL)-6、IL-8、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和结肠组织神经肽五羟色胺(5-hydroxytryptamine,5-HT)、胃饥饿素、血管活性肠肽(vasoactive intestinal peptide,VIP)、P物质(substance P,SP)、神经肽Y(neuropeptide Y,NPY)、降钙素基因相关肽(calcitonin gene-related peptide,CGRY)水平。结果(1)与空白对照组比较,模型组大鼠结肠DAI评分升高(P<0.05);结肠病理损伤明显,可见水肿、充血、炎性浸润,与周围组织粘黏;血清IL-6、IL-8、TNF-α(P<0.05)升高;结肠组织VIP、SP、胃饥饿素、5-HT、NPY水平显著增加,CGRY水平显著降低(P<0.05);(2)与模型组比较,全方组、各拆方组及阳性药组DAI评分降低(P<0.05),溃疡性结肠炎大鼠的病理损伤得到不同程度的减轻。全方组、各拆方组及阳性药组均能降低血清炎症因子IL-6、IL-8、TNF-α水平(P<0.05)。全方组、苦味组、阳性药组大鼠结肠组织VIP、SP、胃饥饿素、5-HT、NPY水平显著降低,CGRY水平显著升高(P<0.05)。酸味组大鼠结肠组织VIP、SP、胃饥饿素、5-HT、NPY降低(P<0.05)。辛味组大鼠结肠组织VIP、胃饥饿素、5-HT降低,SP、CGRY水平显著升高(P<0.05)。甘味组大鼠结肠组织VIP、SP、胃饥饿素、NPY降低,CGRY水平显著升高(P<0.05)。结论乌梅丸及其拆方可能通过调节神经肽和细胞因子这一机制来干预溃疡性结肠炎,全方对神经肽的作用范围更为广泛,其余各拆方组对部分神经肽具有调节作用,各拆方组能够通过协同作用发挥抗炎作用。

Objective To observe the mechanism of fructus mume pills on ulcerative colitis rats,and its compatibility relationship.Methods 83 rats were divided into 8 groups,10 rats in control group,complete receipt group,sour flavour group,bitter flavour group,pungent flavour group,sweet flavour group and SASP group,and 13 rats in model group.UC rats were induced via 4%(v/v)acetic acid enema.The drug was administered 24 hours after molding.Complete rats were orally administrated 900 mg/kg corresponding drug,sour flavour drug group rats were administrated 270 mg/kg,bitter flavour group rats were received 235 mg/kg,pungent flavour group rats were 340mg/kg,sweet group rats were 110mg/kg,and SASP group rats were 300mg/kg.The drug was given intragastric administration at the volume of 10 ml/kg rat body weight,twice a day for seven consecutive days.Control group and model group received equal amount of saline.Disease injury index was used to evaluate UC model,H&E staining were helped to observe pathological damage.Levels of serum inflammatory cytokines IL-6,IL-8,TNF-αand colonic neuropeptide levels of 5-HT,ghrelin,vasoactive intestinal peptide,substance P,neuropeptide Y,calcitonin gene-related peptide were detected by ELISA kits.Results(1)Compared with control group,DAI scores in model group increased(P<0.05).Model group rats colon appeared obvious injuries,including edema,congestion,inflammatory infiltration,and sticking to the surrounding tissue.Serum levels of IL-6,IL-8,TNF-α,were increased(P<0.05),colonic levels VIP,SP,ghrelin,5-HT,NPY increased and CGRY decreased(P<0.05).(2)Compared with model group,DAI scores and colon pathology injuries were decreased in complete receipt group,decomposed group,and SASP group.Serum levels of IL-6,IL-8,TNF-0x0E䥺SymbolaA@0x0F were reduced in complete receipt,modification groups and SASP group.VIP,SP,ghrelin,5-HT,NPY in colon tissue were decreased and CGPY were increased in complete receipt group,bitter flavour group,and SASP group(P<0.05).VIP,SP,ghrelin,5-HT,NPY were decreased in sour flavour group(P<0.05).VIP,ghrelin,5-HT were decreased and SP,CGRY were increased in pungent flavour group(P<0.05).VIP,SP,ghrelin,NPY were reduced and CGRY was raised in sweet group(P<0.05).Conclusion Fructus mume pills and its modifications could treat UC via regulating neuropeptides and inflammation cytokines in Neuro-immune network.The effect of the complete receipt group is obvious,and its modification groups take partial regulation effect on these neuropeptides.All decomposed recipes groups can play anti-inflammatory effects through synergistic effects.