健脾化瘀方对慢性萎缩性胃炎小鼠胃黏膜细胞凋亡和NF-κB/Bcl-2通路的影响

Effects of Jianpi-Huayu Formula on the apoptosis of gastric mucosa and the NF-κB/Bcl-2 pathway in the chronic atrophic gastritis rat model

为探讨健脾化瘀方对慢性萎缩性胃炎(chronic atrophic gastritis,CAG)大鼠胃黏膜细胞凋亡和NF-κB/B淋巴细胞瘤2(B-cell lymphoma 2,Bcl-2)信号通路的影响,将40只SD大鼠随机分为空白组、模型组、阳性组和实验组,每组各10只。除空白组外,其余大鼠利用N-甲基-N’-硝基-N-亚硝基胍(N-methyl-N’-nitro-N-nitrosoguanidine,MNNG)诱导剂结合夹尾刺激和饥饱失常法复制CAG模型。造模成功次日,实验组给予3.0 mL/(kg·d)健脾化瘀方溶液,阳性组给予0.2 g/(kg·d)胃维酶素,空白组和模型组大鼠给予相同体积的生理盐水,灌胃1次/d,连续8周。采用ELISA检测血清胃泌素(gastrin,GAS)、TNF-α和IL-6水平,免疫组化法和Western blotting检测胃黏膜组织中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、NF-κB、Bcl-2蛋白表达,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(terminal deoxynucl neotidyl transferase mediated dUTP nick end labelling,TUNEL)法检测胃黏膜细胞凋亡指数。结果显示,与空白组比较,模型组大鼠血清GAS水平降低(P<0.05),同时血清TNF-α、IL-6水平,胃黏膜细胞凋亡指数,胃黏膜组织PCNA、NF-κB、Bcl-2蛋白表达量增加(P<0.05);而与模型组比较,实验组大鼠血清GAS水平升高(P<0.05),同时血清TNF-α、IL-6水平,胃黏膜细胞凋亡指数,胃黏膜组织PCNA、NF-κB、Bcl-2蛋白表达量降低(P<0.05)。该研究提示,健脾化瘀方可通过抑制NF-κB/Bcl-2信号通路激活调控胃黏膜细胞增殖-凋亡失衡状态,降低炎症反应,进而对CAG小鼠胃黏膜细胞损伤发挥一定的保护作用。

This study aims to explore the effects and molecular mechanism of Jianpi-Huayu Formula on the NF-κB/B-cell lymphoma 2(Bcl-2)pathway in rats with chronic atrophic gastritis(CAG).Forty SD rats were divided into blank group,model group,positive group,and experimental group,with 10 rats in each group.CAG model was established in all except the blank group using tail clamping stimulation and starvation and satiety disorder method after induction with N-methyl-N’-nitro-N-nitrosoguanidine(MNNG).On the next day after successful modeling,the experimental group was given 3.0 mL/(kg·d)Jianpi-Huayu Formula,the positive group was given 0.2 g/(kg·d)vitacoenzyme tablet,while the blank group and the model group were administrated with the same volume of normal saline by gavaging daily for 8 weeks.The levels of serum gastrin(GAS),TNF-α,and IL-6 were detected by ELISA.The expressions of proliferating cell nuclear antigen(PCNA),NF-κB,and Bcl-2 in gastric mucosa were detected by immunohistochemistry and Western blotting.The apoptotic index of gastric mucosal cell was detected by terminal deoxynucl neotidyl transferase mediated dUTP nick end labeling(TUNEL).The results showed that compared to the blank group,the serum GAS level in the model group was decreased(P<0.05),while the serum TNF-α,IL-6 levels,apoptotic index of gastric mucosal cell,and PCNA,NF-κB,Bcl-2 protein expression in gastric mucosa tissue were increased(P<0.05).Meanwhile,compared to the model group,the serum GAS level of the experimental group was increased(P<0.05),while the serum TNF-α,IL-6 levels,apoptotic index of gastric mucosal cell,PCNA,NF-κB,Bcl-2 in gastric mucosa tissue were also decreased(P<0.05).The study suggests that the Jianpi-Huayu Formula can improve the balance of proliferation and apoptosis of gastric mucosal cells by inhibiting activation of NF-κB/Bcl-2 signaling pathway and reducing inflammatory reaction,thus protecting the gastric mucosal cell in CAG rats.