文献类型：期刊文献

中文题名：理中汤合四神丸对脾肾阳虚型溃疡性结肠炎大鼠JNK/Beclin 1/Bcl-2信号通路相关蛋白及基因表达的影响

英文题名：Effects of Lizhong Decoction Combined with Sishen Pills on Expressions of JNK/Beclin 1/Bcl-2 Signaling Pathway Related Proteins and Genes in Ulcerative Colitis Rats with Spleen-kidney Yang Deficiency

作者：郝民琦[1,2];罗蓉[1];王瑞琼[3];田一虹[1];宋耀辉[1];李晓玲[1,2];李海龙[1,4];吴玉泓[1]

第一作者：郝民琦

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]中医方药挖掘与创新转化重点实验室,甘肃兰州730000;[3]兰州现代职业学院,甘肃兰州730000;[4]甘肃中医药大学临床医学院,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2022

卷号：29

期号：1

起止页码：65

中文期刊名：中国中医药信息杂志

外文期刊名：Chinese Journal of Information on Traditional Chinese Medicine

收录：CSTPCD;;CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金(81860810);第四批全国中医基础优秀人才研修项目(J20184832009);甘肃省中医药管理局项目(GZK-2018-5)。

语种：中文

中文关键词：溃疡性结肠炎;温补脾肾法;理中汤合四神丸;自噬;JNK/Beclin 1/Bcl-2信号通路;大鼠

外文关键词：ulcerative colitis;warming and tonifying spleen and kidney;Lizhong Decoction combined with Sishen Pills;autophagy;JNK/Beclin 1/Bcl-2 signaling pathway;rats

摘要：目的观察以温补脾肾法为指导的理中汤合四神丸对脾肾阳虚型溃疡性结肠炎(UC)大鼠的影响,探讨其可能的作用机制。方法制备脾肾阳型UC大鼠模型,将成模大鼠随机分为模型组、柳氮磺吡啶组(SASP组)、肠胃宁组和理中汤合四神丸低、中、高剂量组,每组16只,同时设空白组16只。各给药组予相应药物灌胃,空白组和模型组予等体积蒸馏水灌胃,连续21 d。观察大鼠结肠黏膜组织损伤情况,对结肠黏膜损伤指数(CMDI)进行评分;HE染色观察大鼠结肠组织病理变化;免疫组化和RT-qPCR分别检测大鼠结肠组织JNK1、Beclin 1、Bcl-2、LC3B蛋白和m RNA表达;Western blot检测大鼠结肠组织JNK1、p-JNK1、Bcl-2、p-Bcl-2、Beclin 1和LC3B蛋白表达,并计算LC3BⅡ/LC3BⅠ比值。结果与空白组比较,模型组大鼠CMDI评分显著升高(P<0.01),结肠组织腺体排列紊乱,黏膜层消失,有大量炎性细胞浸润,黏膜基层与基底层分界不清;结肠组织JNK1、Beclin 1、Bcl-2、LC3B mRNA和蛋白表达显著升高(P<0.01),p-JNK1、p-Bcl-2蛋白表达及LC3BⅡ/LC3BⅠ比值显著升高(P<0.01)。与模型组比较,SASP组、肠胃宁组和理中汤合四神丸中、高剂量组大鼠CMDI评分显著降低(P<0.05,P<0.01),各给药组大鼠结肠黏膜损伤减轻,炎性细胞浸润减少;理中汤合四神丸高剂量组、SASP组、胃肠宁组大鼠结肠组织JNK1、Beclin 1、Bcl-2、LC3B mRNA和蛋白表达显著降低(P<0.01),p-JNK1、p-Bcl-2蛋白表达及LC3BⅡ/LC3BⅠ比值显著降低(P<0.01)。结论以温补脾肾法为指导的理中汤合四神丸可能通过抑制JNK/Beclin 1/Bcl-2信号通路的激活,降低结肠组织过度自噬,进而修复脾肾阳虚型UC大鼠结肠黏膜。
Objective To observe the effects of warming and tonifying spleen and kidney method on guiding Lizhong Decoction combined with Sishen Pills in ulcerative colitis(UC)rats with spleen-kidney yang deficiency;To discuss its possible mechanism of action.Methods UC model of spleen-kidney yang deficiency was established.The model rats were randomly divided into model group,sulfasalazine group,Changweining group,Lizhong Decoction combined with Sishen Pills low-,medium-and high-dosage groups,with 16 in each group,and 16 in blank group.The blank group and model group were given distilled water by gavage,and the treatment groups were given corresponding drugs for 21 consecutive days.Rat colonic mucosal tissue damage was observed and the colonic mucosal damage index(CMDI)was scored.HE staining was used to observe the pathological changes of rat colon tissue;immunohistochemistry and RT-qPCR were used to detect JNK1,Beclin 1,Bcl-2,LC3 B protein and mRNA expression in rat colon tissue;Western blot was used to detect the protein expressions of JNK1,p-JNK1,Bcl-2,p-Bcl-2,Beclin 1,and LC3 B in rat colon tissue,and the ratio of LC3 BⅡ/LC3 BⅠwas calculated.Results Compared with the blank group,the CMDI score of the model group significantly increased(P<0.01),the glands of the colon tissue were arranged disorderly,the mucosal layer disappeared,a large number of inflammatory cells infiltrated,the basal mucosa and the basal layer were unclear,and the expression of JNK1,Beclin 1,Bcl-2 and LC3 B mRNA and protein increased significantly(P<0.01)in the colon tissue,the proteins expression of p-JNK1 and p-Bcl-2 and the ratio of LC3 BⅡ/LC3 BⅠincreased significantly(P<0.01).Compared with the model group,the CMDI score of the rats in the SASP group,Changweining group and Lizhong Decoction combined with Sishen Pills medium-and high-dosage groups was significantly decreased(P<0.05,P<0.01),and the colonic mucosal damage of the rats in each administration group was reduced,the inflammatory cell infiltration decreased.The expressions of JNK1,Beclin 1,Bcl-2,LC3 B mRNA and protein in the colon tissue of the Lizhong Decoction combined with Sishen Pills high-dosage group,SASP group,and Changweining group were significantly decreased(P<0.01),the expressions of p-JNK1 and p-Bcl-2 protein and the ratio of LC3 BⅡ/LC3 BⅠwere significantly reduced(P<0.01).Conclusion Lizhong Decoction combined with Sishen Pill guided by the method of warming and tonifying spleen and kidney may inhibit the activation of JNK/Beclin 1/Bcl-2 signaling pathway,avoid excessive autophagy of colon tissue,and then repair the colon mucosa of UC rats with spleen-kidney yang deficiency.