摘要
目的基于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路探讨化浊解毒方干预慢性萎缩性胃炎(CAG)大鼠的作用机制。方法采用水杨酸钠、N-甲基-N’-硝基-N-亚硝基胍(MNNG)以及饥饱失常多因素诱导大鼠模型。将造模成功的50只大鼠按随机数字表法分为模型组,摩罗丹组,化浊解毒高、中、低剂量组,每组10只。同时另设正常组10只。化浊解毒方高、中、低剂量组分别灌以化浊解毒方(36、18、9 g/kg),摩罗丹组灌以摩罗丹1.4 g/kg,正常组和模型组灌同等体积生理盐水,均干预60天。苏木素-伊红(HE)观察胃黏膜组织形态学变化;免疫比浊法检测大鼠血清胃蛋白酶原Ⅰ(PGⅠ)、PGⅡ以及计算PGⅠ/PGⅡ(PGR)比值;放射免疫法检测大鼠血清胃泌素17(G-17)水平;Western Blot检测胃黏膜PI3K、p-AKT蛋白表达含量;RT-qPCR检测PI3K、AKT、人第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)、Bcl-2相关凋亡促进因子(BAD)、细胞抗凋亡因子B淋巴细胞瘤-2(Bcl-2)mRNA水平。结果与正常组比较,模型组胃黏膜组织固有腺体萎缩,排列混乱数量减少,炎性细胞浸润;血清G-17、PGⅠ、PGR及胃组织PTEN mRNA降低(P<0.05),血清PGⅡ、胃组织PI3K、p-Akt蛋白升高(P<0.05),PI3K、Akt、Bad、Bcl-2 mRNA表达亦升高(P<0.05)。与模型组比较,各药物干预组病理情况均有所改善,其中化浊解毒方高、中剂量组对胃黏膜炎症及萎缩改善明显(P<0.05);化浊解毒方各剂量组和摩罗丹组血清G-17、PGⅠ升高,Bad、Bcl-2 mRNA表达降低(P<0.05);化浊解毒方高、中剂量组和摩罗丹组PGR值、PTEN mRNA升高,PI3K、p-AKT蛋白及mRNA降低(P<0.05)。与摩罗丹组比较,化浊解毒方高、中剂量组G-17、PGⅠ升高,Bad、Bcl-2 mRNA降低,化浊解毒方高剂量组PI3K mRNA降低(P<0.05)。结论化浊解毒方对CAG大鼠有干预作用,其作用机制可能与抑制PI3K/AKT信号的激活,促进胃黏膜细胞凋亡,从而发挥保护胃黏膜作用。
Objective Based on the signal pathway of phosphatidylinositol 3-kinase(PI3K)/protein kinase B protein kinase B(AKT),to explore the mechanism of Huazhuo Jiedu Recipe(HZJDR)on chronic atrophic gastritis(CAG)rats.Methods The rat model was induced by sodium salicylate,N-methyl-nitronitro-N-nitrosoguanidine(MNNG)and hunger and satiety disorder.Fifty rats were randomly divided into model group,morodan group and high,medium and low dose HZJDR groups of removing turbidity and detoxification,with 10 rats in each group.At the same time,10 rats in normal group were set up.The high,middle and low dose HZJDR groups were administered with HZJDR(36,18,9 g/kg)by gastrogavage,respectively.The control group was administered with morodan 1.4 g/kg.Equal volume of normal saline was administered to rats in the normal group and model group by gastrogavage.All rats were intervened for 60 days.The histomorphological changes of gastric mucosa were observed by HE method,the levels of serum pepsinogenⅠ(PGⅠ)and PGⅡwere detected by immunoturbidimetry,the ratio of PGⅠ/PGⅡ(PGR)was calculated,the level of serum gastrin 17(G-17)was detected by radioimmunoassay.The protein expression of PI3K and p-AKT in gastric mucosa was detected by Western Blot,and the levels of PI3K,AKT,PTEN,Bad and Bcl-2mRNA were detected by RT-qPCR.Results Compared with the normal group,the inherent glands of gastric mucosa atrophied,the number of disordered arrangement decreased,and inflammatory cells infiltrated.Serum G-17,PGⅠ,PGR and PTEN mRNA in gastric tissue decreased,while serum PGⅡ,PI3K and p-Akt protein in gastric tissue increased(P<0.05);the expressions of PI3K,Akt,Bad,and Bcl-2 mRNA were also increased(P<0.05).Compared with the model group,the pathological conditions of each drug intervention group were improved,and the high and medium dose HZJDR groups significantly improved the inflammation and atrophy of gastric mucosa(P<0.05);serum G-17 and PGⅠincreased,the expression of Bad and Bcl-2 mRNA decreased in each dose HZJDR group and morodan group(P<0.05);PGR and PTEN mRNA increased,PI3K,p-Akt protein and mRNA decreased in high and medium dose HZJDR groups and morodan group(P<0.05).Compared with morodan group,G-17 and PGⅠincreased,Bad and Bcl-2 mRNA decreased in high and medium dose HZJDR groups,and PI3K mRNA decreased in high dose HZJDR groups(P<0.05).Conclusion HZJDR has an interventive effect on CAG rats,and its mechanism may be related to inhibiting the activation of PI3K/Akt signal and promoting the apoptosis of gastric mucosal cells,so as to protect gastric mucosa.
作者
王杰
马虹宇
高云霄
贾雪梅
郭瑜西
杨柳
李泽
李博林
杨倩
WANG Jie;MA Hong-yu;GAO Yun-xiao;JIA Xue-mei;GUO Yu-xi;YANG Liu;LI Ze;LI Bo-lin;YANG Qian(Department of Spleen and Stomach,Hebei University of Chinese Medicine,Shijiazhuang 050091;Department of Traditional Chinese Medicine,Hebei General Hospital,Shijiazhuang 050051;Department of Spleen and Stomach,Hebei Province Hospital of Chinese Medicine,Shijiazhuang 050011)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2022年第3期348-354,共7页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家中医临床研究基地建设项目(No.国中医药办科技函[2018]18号)
国家科技部重点研发课题(No.2018YFC1704100,No.2018YFC1704102)
河北省省级科技计划资助(No.21377724D,No.21377740D)
河北省中医药管理局科研计划项目(No.2021034)。
