健脾清热活血方干预PI3K-Akt/mTOR信号通路调控CDK1表达防治溃疡性结肠炎相关癌变

Jianpi Qingre Huoxue Formula Intervenes PI3K-Akt/mTOR Signaling Pathway and Regulation CDK1 Expression and Prevention to Control Collapse Related Canceration

目的:观察健脾清热活血方对溃疡性结肠炎相关癌变小鼠组织形态、超微结构、PI3K-Akt/mTOR信号通路中相关指标及CDK1的表达变化,探讨其防治溃结相关癌变的可能机制。方法:140只SPF级Balb/c小鼠按体质量随机分为正常组、模型组、阻断剂组、治疗组及对照组,每组20只;除外正常组,其余各组采用DMH/DSS复合法制备溃结癌变模型,模型组予等剂量生理盐水,阻断剂组予PI3K阻断剂LY294002干预,治疗组予不同剂量健脾清热活血方药,对照组予美沙拉嗪,连续干预16周。应用光镜及电镜观察溃结癌变小鼠结肠组织形态学及超微结构变化;Western-blot及Real-time PCR检测结肠黏膜P110、P85、P-AKT、mTOR、CDK1蛋白及其mRNA表达变化。结果:模型组见黏膜上皮脱落,溃疡形成,腺体萎缩,部分有浸润癌表现,治疗组所见黏膜损伤程度较模型组有不同程度改善,其中以中、高剂量组结肠黏膜损伤改善情况显著,部分可见黏膜充血并肿胀,不典型增生及淋巴滤泡出现(P<0.05)。与正常组比较,模型组P110、P85、CDK1蛋白表达量上升(P<0.05);与模型组比较,治疗组P85、CDK1蛋白表达下降(P<0.05)。与模型组比较,治疗组P85、P-Akt基因表达下调(P<0.05)。各组间P110、mTOR基因表达量比较差异无统计学意义(P>0.05)。结论:健脾清热活血方可能通过下调P85、P-Akt,介导CDK1表达,诱导炎症缓解,修复肠道黏膜,发挥防治UCAC的效用。

Objective ：To observe Jianpi Qingre Huoxue formula for the Organization form ,Ultrastructure and The relevant indi- cators of PI3K - Akt/mTOR signaling pathway which parts of the mice that ulcerative colitis associated cancer and the expres- sion of CDK1 changes. Discuss the possible mechanism for prevention and control of the collapse related canceration. Methods ： 140 SPF Balb/c mice are equally divided into normal group, model group, blocker group, intervention group and control group by weight. Except normal group,the other groups using the DMH/DSS methods to make ulcerative colitis associated carcinogenesis model. The model group was treated with the same dose of normal saline, and the blocker group was treated with PI3K blocker LY294002. The intervention group was treated with different doses of Jianpi Qingre Huoxue formula, while the control group was treated with mesalazine for 16 weeks. Using light microscope and electron microscope, we observed the morphological and ultra- structural changes of UCAC, and the Protein and mRNA expression of P110, P85, P - AKT, roTOR and CDK1 in colonic mucosa were detected by Real - time PCR and Western - blot. Results ： The model group is characterized by epithelial exfoliation, ulcer for- mation, gland atrophy, some invasive carcinoma. However, the intervention group is characterized by mucosal hyperemia and ede- ma, atypical hyperplasia and lymph follicles. The extent of gastric mucosal lesions were improved compared with model group, es- pecially in the middle and high dose group obviously. Compared with the normal group, the expression of P110, P85 and CDK1 protein in the model group were in the inereasment （ P 〈 O. 05 ）. Compared with the model group, the expression of P85 and CDK1 in the intervention group were on the decline （P 〈 0. 05）. There was no significant difference in the expression of P110 androTOR between each group （ P 〉 0. 05 ）. Conclusion ： The Jianpi Qingre Huoxue formula play a role in the prevention and treatment of UCAC by reducingthe expression of P85 , P - Akt and mediating the expression of CDK1 which improve inflammation and re- pair intestinal mueosa.