摘要
目的从细胞骨架破坏、炎症反应和缺氧环境角度开展清肠温中方的拆方研究,阐明该方在溃疡性结肠炎(ulcerative colitis, UC)治疗中,针对不同病机更加有效的药物组成单元,为其药物配伍和临床应用提供科学依据。方法 79只SD大鼠随机分10组,其中空白组7只,模型组、温中健脾组、清热燥湿组、凉血化瘀组、温中健脾合清热燥湿组、温中健脾合凉血化瘀组、清热燥湿合凉血化瘀组、全方组和美沙拉秦组,每组8只。除空白组外,其余大鼠连续7天自由饮用4%葡聚糖硫酸钠(dextran sulfate sodium,DSS)溶液以建立急性UC模型。随后7天按不同分组灌胃给药,与此同时,除空白组外的所有大鼠,仍继续饮用1%DSS溶液以维持小剂量刺激引起的病变。通过组织学病变评分、结肠低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)蛋白电泳、波形蛋白(vimentin,VIM)和角蛋白8(cytokeratin 8,K8)免疫组化染色、血清干扰素γ诱导蛋白10(interferon-γ-inducible protein 10,IP10)、趋化因子受体3(chemokine receptor 3,CXCR3)Elisa检测,开展清肠温中方拆方的拆方研究。结果与模型组相比,清热燥湿、温中健脾合清热燥湿、温中健脾合凉血化瘀和全方组能明显降低组织学病变评分(P<0.05)。全方及拆方各组均能降低结肠中VIM/K8蛋白表达比值,其中以温中健脾合凉血化瘀组效果更明显(P<0.05)。全方及拆方各组均能减少血清中IP10含量,温中健脾合凉血化瘀和温中健脾合清热燥湿组效果更为明显(P<0.05);除凉血化瘀组外各组均能降低血清CXCR3含量,其中以清热燥湿合凉血化瘀和温中健脾合凉血化瘀组降低更多,且联合组疗效优于美沙拉秦(P<0.05)。全方及拆方各组均能减少结肠中HIF-1α表达,以温中健脾合凉血化瘀、温中健脾合清热燥湿和全方组的下降趋势更明显,但各组间不存在统计学差异。结论清肠温中方全方的作用优于温中健脾合凉血化瘀和温中健脾合清热燥湿组,而两组又比单用温中健脾、清热燥湿或凉血化瘀有更明显的调控作用。进一步证实了在UC的治疗中,清热燥湿或凉血化瘀的祛邪治法,只有在联合了温中健脾的扶正药物后,才能更好地发挥修复细胞骨架、减轻炎症反应和改善缺氧环境的作用。提示在UC的临床治疗中,需要重视顾护正气,即温中健脾,才能取得更好的疗效。
Objective To carry out the separate components study of Qingchang Wenzhong formula from the perspective of cytoskeleton destruction,inflammatory reaction and hypoxia environments.To clarify that the prescription can make drug units composed of more effective drugs for different pathogenesis in the treatment of ulcerative colitis(UC),and to provide scientific basis for drug compatibility and clinical application.Methods 79 SD rats were randomly divided into 10 groups,including 7 in the blank group,8 in each group,including model group,Wenzhong Jianpi group,Qingre Zaoshi group,Liangxue Huayu group,Wenzhong Jianpi combined with Qingre Zaoshi group,Wenzhong Jianpi combined with Liangxue Huayu group,Qingre Zaoshi combined with Liangxue Huayu group,Qingchang Wenzhong formula group and mesalazine group.Except the blank group,the other rats were free to drink 4% dextran sulfate sodium(DSS) solution for 7 days to establish acute UC rats model.The drug was administered by gavage in different groups over the next 7 days.At the same time,all rats except the blank group continued to drink 1% DSS solution to maintain the lesions caused by low doses of stimulation.Scoring by histological lesion,hypoxia inducible factor-1α(HIF-1α),protein electrophoresis,vimentin(VIM) and cytokeratin8(K8) were detected by immunohistochemical staining.Interferon-γ-inducible protein 10(IP10) and chemokine receptor 3(CXCR3) detected by ELISA.All these methods were used to carry out separate components study of Wenchang Qingzhong formula.Results Compared with the model group,the score of pathological changes in Qingre Zaoshi group,Wenzhong Jianpi combined with Qingre Zaoshi group,Wenzhong Jianpi combined with Liangxue Huayu group and Wenchang Qingzhong formula group reduced significantly(P <0.05).The ratio of VIM/K8 protein expression in colon decreased in Wenchang Qingzhong formula group and other separate components groups,and the Wenzhong Jianpi combined with Qingre Zaoshi group decreased more obviously(P<0.05).The content of IP10 in serum decreased in Wenchang Qingzhong formula group and other separate components groups,and the Wenzhong Jianpi combined with Liangxue Huayu group and Wenzhong Jianpi combined with Qingre Zaoshi group decreased more obviously(P<0.05).Except the Liangxue Huayu group,the content of CXXR3 in serum decreased in other separate components group,and Qingre Zaoshi combined with Liangxue Huayu group and Wenzhong Jianpi combined with Liangxue Huayu group decreased more obviously,and the combined group’s curative effect is better than mesalazine group(P <0.05).The HIF-la expression in colon decreased in Wenchang Qingzhong formula group and other separate components groups,and the Wenzhong Jianpi combined with Liangxue Huayu group and Wenzhong Jianpi combined with Qingre Zaoshi group and Wenchang Qingzhong formula group decreased more obviously,but there was no statistical difference between the groups.Conclusion The curative effect of Qingchang Wenzhong formula group is better than Wenzhong Jianpi combined with Liangxue Huayu group and Wenzhong Jianpi combined with Qingre Zaoshi group,and two combined group’s regulating effect is obviously better than single Wenzhong Jianpi or Qingre Zaoshi or Liangxue Huayu group.It is further confirmed that in the treatment of UC,the method of removing pathogenic factors such as clearing away heat,drying dampness or cooling blood and removing blood stasis can play a better role in repairing cytoskeleton,alleviating inflammatory reaction and improving hypoxia environment only after the combination of strengthening vital qi drugs for warming and strengthening the spleen and stomach.It is suggested that in the clinical treatment of UC,attention should be paid to the care of vital qi,that is,warming and strengthening the spleen and stomach,so as to achieve better curative effect.
作者
石磊
李军祥
韩啸
刘佳丽
路琼琼
寇富舜
孙中美
丁庞华
毛堂友
王志斌
SHI Lei;LI Junxiang;HAN Xiao;LIU Jiali;LU Qiongqiong;KOU Fushun;SUN Zhongmei;DING Panghua;MAO Tangyou;WANG Zhibin(School of Life Sciences,Beijing University of Chinese Medicine,Beijing,100029,China)
出处
《环球中医药》
CAS
2020年第7期1134-1141,共8页
Global Traditional Chinese Medicine
基金
2018年国家重点研发计划(2018YFC1705403,2018YFC1705405)
2018年中医药管理局“百千万”人才工程岐黄学者(李军祥)项目
2019年北京中医药大学研究生自主选题(2019-JYB-XS-209)。
关键词
溃疡性结肠炎
清肠温中方
细胞骨架
缺氧环境
炎症反应
拆方研究
大鼠
Ulcertaive colitis
Qingchang Wenzhongformula
Cytoskeleton
Hypoxia environment
Inflammatory reaction
Separate components study
Rats
作者简介
石磊(1989-),博士,博士后。研究方向:中西医结合防治慢性胃肠病。E-mail:klmysl1205@126.com;通信作者:王志斌(1978-),博士,副主任医师。研究方向:中西医结合防治慢性胃肠病。E-mail:wangsanger@126.com。
