健脾清化化瘀汤对幽门螺杆菌相关胃癌前病变大鼠胃黏膜miR-21、PTEN表达的影响  

Effect of Jianpi Qinghua Huayu decoction on the expression of miR-21 and PTEN in gastric mucosa of rats with HP related gastric precancerous lesions

作　　者：李一桐 汪红兵[2] LI Yitong;WANG Hongbing(The Second Clinical Medical College of Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学第二临床医学院,100078 [2]首都医科大学附属北京中医医院脾胃病科

出　　处：《环球中医药》2023年第11期2153-2160,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81774257)。

摘　　要：目的探讨健脾清化化瘀汤治疗胃癌前病变的可能作用机制。方法将90只健康清洁级雄性SD大鼠按照随机数字表法随机分为6组,包括正常组、模型组、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)抑制剂组、健脾清化化瘀汤低剂量组、健脾清化化瘀汤中剂量组、健脾清化化瘀汤高剂量组。除正常组外,其余5组大鼠均用1×10^(9)CFU/mL幽门螺杆菌(helicobacter pylori,HP)菌液灌胃7次,平均2次/周。确认HP感染后,除正常组外,其余5组均予150μg/mL甲基-N-硝基-N亚硝基胍(MNNG)溶液灌胃,频率5次/周,同时予100μg/mL MNNG溶液自由饮用,配合0.03%雷尼替丁饲料喂养、饥饱失常法(2天足量饲料喂养,1天禁食),持续造模28周。造模成功后,健脾清化化瘀汤低、中、高剂量组分别给予相应剂量健脾清化化瘀汤灌胃,iNOS抑制剂组给予氨基胍溶液50 mg/(kg·d)剂量进行灌胃,正常组及模型组予以生理盐水进行灌胃,均1次/天,干预治疗10周。干预结束后,运用苏木精—伊红染色观察大鼠胃黏膜病理变化;运用免疫组化法检测胃组织磷酸酯酶与张力蛋白同源物(phosphate and tension homology deleted on chromosome ten,PTEN)蛋白表达;运用RT-PCR法检测胃组织miR-21、PTEN mRNA表达。结果与正常组比较,模型组大鼠胃黏膜可见腺体萎缩、肠化、异型增生,健脾清化化瘀汤中、高剂量组及iNOS抑制剂组较模型组有不同程度的改善,健脾清化化瘀汤低剂量组改善不明显。各组大鼠胃黏膜miR-21、PTEN mRNA、PTEN蛋白表达差异均有统计学意义(P<0.01)。与空白组比较,模型组miR-21表达显著升高(P<0.01),PTEN mRNA、蛋白表达显著下调(P<0.01;P<0.01);与模型组比较,健脾清化化瘀汤中、高剂量组、iNOS抑制剂组miR-21表达下调(均P<0.01),PTEN mRNA、蛋白表达升高(P<0.05;P<0.01),健脾清化化瘀汤低剂量组miR-21、PTEN mRNA、蛋白表达较模型组差异均无统计学意义。结论健�Objective To explore the possible mechanism of Jianpi Qinghuahuayu decoction in the treatment of gastric precancerous lesions.Methods 90 male SD rats were randomly divided into 6 groups according to the random number table method,including normal group,model group,inducible nitric oxide synthase(iNOS)inhibitor group,Jianpi Qinghua Huayu decoction low-dose group,Jianpi Qinghua Huayu decoction middle-dose group,Jianpi Qinghua Huayu decoction high-dose group.Except the normal group,rats in the other 5 groups were gavaged with 1×10^(9)CFU/mL HP bacteria liquid 7 times,an average of 2 times per week.After confirming HP infection,except the normal group,the other 5 groups were given 150μg/mL MNNG solution by gavage at a frequency of 5 times/week.At the same time,they were given 100μg/mL MNNG solution for free drinking and fed with 0.03%ranitidine feed.Satisfaction method(2 day feeding with adequate feed,1day fasting),continued modeling for 28 weeks.After modeling,the low,medium and high-dose groups of Jianpi Qinghua Huayu decoction were given corresponding doses of Jianpi Qinghua Huayu decoction by gavage,and the iNOS inhibitor group was given aminoguanidine solution 50 mg/(kg·d).Gavage,the normal group and the model group were given normal saline gavage,both once a day,intervention treatment for 10 weeks.After the intervention,the method of HE staining was used to observe the pathological changes of the gastric mucosa of rats;the expression of PTEN protein in gastric tissue was detected by immunohistochemical method;the expression of miR-21 and PTEN mRNA in gastric tissue were detected by RT-PCR method.Results Compared with the normal group,the gastric mucosa of the model group showed gland atrophy,intestinal metaplasia,and dysplasia.The middle and high-dose Jianpi Qinghua Huayu decoction group and the iNOS inhibitor group had different degrees of improvement compared with the model group.The low-dose Jianpi Qinghua Huayu decoction group did not improve.The differences among the expression of miR-21,PTEN mRNA and

关 键 词：健脾清化化瘀汤 MIR-21 慢性萎缩性胃炎伴异型增生 磷酸酯酶与张力蛋白同源物 胃癌前病变 

分 类 号：R285.5[医药卫生—中药学]