浏览全部资源
扫码关注微信
湖北民族大学,湖北 恩施 445000
Published:20 July 2023,
Published Online:16 February 2023,
Received:07 October 2022,
扫 描 看 全 文
谢慧臣,冉云,张云等.生品与麸炒苍术对脾虚证大鼠小肠吸收转运相关蛋白载体的影响[J].中国实验方剂学杂志,2023,29(14):47-56.
XIE Huichen,RAN Yun,ZHANG Yun,et al.Effect of Raw and Bran-fried Atractylodis Rhizoma on Protein Carriers Related to Small Intestinal Absorption Transport in Spleen Deficiency Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(14):47-56.
谢慧臣,冉云,张云等.生品与麸炒苍术对脾虚证大鼠小肠吸收转运相关蛋白载体的影响[J].中国实验方剂学杂志,2023,29(14):47-56. DOI: 10.13422/j.cnki.syfjx.20230602.
XIE Huichen,RAN Yun,ZHANG Yun,et al.Effect of Raw and Bran-fried Atractylodis Rhizoma on Protein Carriers Related to Small Intestinal Absorption Transport in Spleen Deficiency Rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(14):47-56. DOI: 10.13422/j.cnki.syfjx.20230602.
目的
2
通过比较生品与麸炒苍术对脾虚证大鼠小肠组织结构及转运相关蛋白载体的影响,探讨麸炒法影响苍术改善胃肠功能的可能机制。
方法
2
雄性SD大鼠70只随机分为正常组、脾虚证模型组(以下简称模型组)、生苍术高、低剂量组、麸炒苍术高、低剂量组、复方谷氨酰胺组,每组10只。除正常组外的6组均以苦寒破气加饥饱失常法复制脾虚模型,造模时间为21 d。完成造模后,各治疗组每天按相应剂量药物灌胃治疗,共计14 d:生苍术高、低剂量组10、2.5 g·kg
-1
;麸炒苍术高、低剂量组10、2.5 g·kg
-1
,复方谷氨酰胺组9 mg·kg
-1
;正常组、模型组给予同体积生理盐水灌胃。治疗期间行脾虚大鼠一般生存状况、宏观证候评分及每日体质量增加量和摄食量、肛温测定,治疗期满后处死大鼠,苏木素-伊红(HE)染色观察各组大鼠小肠组织病理学改变,酶联免疫吸附测定法(ELISA)检测血清5-羟色胺(5-HT)含量,比色法检测血清
D
-木糖、乳酸、淀粉酶含量,考马斯亮蓝(Bradford)法检测小肠组织游离脂肪酸受体3(FFA3)、肽转运载体1(PepT1)含量水平,免疫组化法检测小肠组织中钠依赖性葡萄糖转运体1(SGLT1)、葡萄糖转运体1(GLUT1)蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测葡萄糖转运体2(GLUT2)、钠/氢交换因子3(NHE3)、5-羟色胺受体4(5-HT4R) mRNA表达。
结果
2
与正常组比较,模型组大鼠造模结束时,大部均表现为精神迟钝,眯眼食少,倦怠少动等脾虚证表现,小肠黏膜上皮及固有膜内基本结构被破坏,血清乳酸、5-HT含量升高,血清淀粉酶及
D
-木糖含量降低(
P
<
0.01),与模型组比较,各个治疗组一般表现不同程度好转,小肠微观结构不同程度修复,体质量每日增加量、肛温及脾虚宏观证候积分不同程度好转(
P
<
0.05,
P
<
0.01),血清乳酸、5-HT含量不同程度降低,血清淀粉酶及
D
-木糖含量不同程度升高(
P
<
0.05,
P
<
0.01),小肠组织中PepT1含量不同程度升高,FFA3含量不同程度降低,SGLT1、GLUT1蛋白表达不同程度升高,GLUT2、NHE3 mRNA表达不同程度升高,5-HT4R mRNA表达不同程度降低(
P
<
0.05,
P
<
0.01);与生苍术高、低剂量组比较,麸炒苍术高、低剂量组一般表现及小肠组织病理学改善更为明显,体质量每日增加量、肛温、脾虚宏观证候积分亦有明显改善(
P
<
0.05,
P
<
0.01);血清乳酸、5-HT含量明显降低,血清淀粉酶及
D
-木糖含量明显升高(
P
<
0.05,
P
<
0.01);小肠组织中PepT1含量明显升高,FFA3含量明显降低,SGLT1、GLUT1蛋白表达明显升高,GLUT2、NHE3 mRNA表达明显升高,5-HT4R mRNA表达明显降低(
P
<
0.05,
P
<
0.01)。
结论
2
麸炒苍术可更好地改善脾虚模型大鼠脾虚相关证候及小肠组织病理学,其机制可能与苍术麸炒后可更好地调节小肠转运相关蛋白载体的表达及各种消化酶的分泌,恢复小肠吸收转运功能有关。
Objective
2
To explore the underlying mechanism of bran-fried Atractylodis Rhizoma (AR) in improving gastrointestinal function by comparing the effects of raw AR and bran-fried AR on the small intestine tissue structure and transport-related protein carriers in rats with spleen deficiency syndrome.
Method
2
Seventy male SD rats were randomly divided into a normal group, a model group, high- and low-dose raw AR groups (10, 2.5 g·kg
-1
), high- and low-dose bran-fried AR groups (10, 2.5 g·kg
-1
), and a compound glutamine group (9 mg·kg
-1
), with 10 rats in each group. Except for the normal group, the other six groups were subjected to the spleen deficiency model induced by the method of bitter and cold breaking stagnated Qi and abnormal hunger and fullness for 21 days. After modeling, each treatment group was given medication orally according to the corresponding doses every day for a total of 14 days, and the normal group and the model group were given an equal volume of normal saline orally. During the treatment period, the general survival status, macroscopic syndrome score, daily increase in body weight and food intake, and rectal temperature of the spleen deficiency rats were evaluated, and after the treatment, the rats were sacrificed. The pathological changes in the small intestine tissues of each group were observed by hematoxylin-eosin (HE) staining. The content of serum 5-hydroxytryptamine (5-HT) was detected by enzyme-linked immunosorbent assay (ELISA), and the content of serum
D
-xylose, lactate, and amylase was detected by colorimetry. The levels of free fatty acid receptor 3 (FFA3) and peptide transporter 1 (PepT1) in small intestinal tissues were detected by the Bradford method, and the protein expression of sodium-dependent glucose transporter 1 (SGLT1) and glucose transporter 1 (GLUT1) in small intestinal tissue was detected by immunohistochemistry. Real-time fluorescence-based quantitative PCR was used to detect the mRNA expression of glucose transporter 2 (GLUT2), sodium/hydrogen exchanger 3 (NHE3), and 5-hydroxytryptamine receptor 4 (5-HT4R).
Result
2
Compared with the normal group, the model group exhibited symptoms of spleen deficiency, such as sluggishness, squint, reduced food intake, and lethargy at the end of modelling, damaged basic structure of the small intestinal mucosal epithelium and lamina propria, increased serum lactate and 5-HT content, and decreased serum amylase and
D
-xylose (
P
<
0.01). Compared with the model group, all treatment groups showed varying degrees of improvement, with the small intestinal microstructure repaired to different degrees. The daily weight gain, anal temperature, and macroscopic syndrome score of spleen deficiency improved to varying degrees (
P
<
0.05,
P
<
0.01). The serum lactate and 5-HT content decreased to varying degrees, while the serum amylase and
D
-xylose content increased to varying degrees (
P
<
0.05,
P
<
0.01). The PepT1 content in the small intestinal tissues increased, while the FFA3 content decreased to varying degrees. The protein expression of SGLT1 and GLUT1 increased, while the mRNA expression of GLUT2 and NHE3 increased to varying degrees. The mRNA expression of 5-HT4R decreased to varying degrees (
P
<
0.05,
P
<
0.01). Compared with the high- and low-dose raw AR groups, the high- and low-dose bran-fried AR groups showed significant improvement in general conditions and histopathological improvement of the small intestinal tissues. The daily weight gain, anal temperature, and macroscopic syndrome score of spleen deficiency also improved (
P
<
0.05,
P
<
0.01). The serum lactate and 5-HT content decreased, while the serum amylase and
D
-xylose content increased (
P
<
0.05,
P
<
0.01). The PepT1 content in the small intestinal tissues increased, while the FFA3 content decreased. The protein expression of SGLT1 and GLUT1 increased, while the mRNA expression of GLUT2 and NHE3 increased. The mRNA expression of 5-HT4R decreased significantly (
P
<
0.05,
P
<
0.01).
Conclusion
2
Bran-fried AR can improve the spleen deficiency-related symptoms and histopathology of the small intestinal tissues in spleen deficiency model rats. Its mechanism may be related to the regulation of the expression of various transport-related protein carriers and the secretion of various digestive enzymes after stir-frying of AR, thus restoring the absorption and transport function of the small intestine.
麸炒苍术脾虚钠依赖性葡萄糖转运体、葡萄糖转运体信号通路
bran-fryingAtractylodis Rhizomaspleen deficiencysodium-dependent glucose transporter, glucose transportersignaling pathway
宋梧桐.中医脾虚证病机演变过程及辨证论治研究[D].天津:天津中医药大学,2021.
于艳,袁媛,贾天柱,等.苍术炮制前后化学成分及药理作用研究近况[J].时珍国医国药,2016,27(1):189-191.
胡小勤,曾雪霞,付蓉,等.黄芩、苍术对脾胃湿热证大鼠燥湿作用的性效关系[J].中国实验方剂学杂志,2021,27(13):35-42.
瞿领航,刘艳菊,涂济源,等.苍术燥性探讨及研究展望[J].中国实验方剂学杂志,2018,24(20):217-222.
何泽正,范仕嘉,许开武,等.小肠上皮葡萄糖转运蛋白表达和功能异常的影响因素[J].肠外与肠内营养,2021,28(6):376-380.
余玉英.脂肪组织特异性nSREBP1c表达下小肠糖吸收功能研究及FTZ干预[D].广州:广东药科大学,2019.
王琦越.平胃散对湿阻中焦证大鼠十二指肠水糖协同转运蛋白影响的研究[D].成都:成都中医药大学,2017.
秦生发,张铮,蓝敏敏,等.逍遥散对慢性束缚应激肝郁脾虚证大鼠MMP2、TIMP2表达的影响[J].中国中医基础医学杂志,2021,27(7):1084-1088.
成映霞,杜娟,杨晓轶,等.脾气虚大鼠小肠组织Ca2+/CaM信号通路关键分子的表达变化[J].中华中医药杂志,2015,30(10):3624-3628.
郑彼萸.中药新药临床研究指导原则(试行)[M].北京:中国科技出版社,2002:361-364.
郑小伟.中医实验动物模型方法学[M].上海:上海中医药人学出版社,1999:63-111.
蒋琦,彭怡雯,沈淑怡,等.脾虚证动物模型的研究进展[J].基层医学论坛,2021,25(13):1903-1905.
黄辰,李瑛,赵妍,等.脾虚证动物模型评价方法评述[J].辽宁中医杂志,2018,45(2):433-437,后插3.
蒋萃,李白雪,袁世清,等.脾虚证大鼠模型的制备方法研究进展及评价[J].环球中医药,2018,11(8):1333-1336.
卢梦雄,黄金科,王一帆,等.高乳糖饮食叠加水平台法脾虚证模型研究与评价[J].世界中医药,2022,17(21):3033-3038.
李亚欢,张冬梅,王淑艳,等.脾虚证动物模型研制概况[J].中医药导报,2019,25(1):100-102.
郑京胜,金利思,王孙富,等.中药炮制辅料麦麸的研究思路探析[J].中华中医药杂志,2019,34(2):680-683.
于欢,刘德文,龚鹏飞,等.苍术炮制方法及其饮片的质量控制、药效评价研究进展[J].中国实验方剂学杂志,2017,23(23):194-200.
章万君.参苓白术散对脾虚泄泻大鼠脑-肠轴5-HT的调控作用[D].合肥:安徽农业大学,2021.
张明,吴晶,彭真,等.荷泽颗粒对脾虚大鼠血清胃泌素、D-木糖浓度、磷酸肌酸激酶的影响[J].中国中医急症,2020,29(2):267-269,277.
李元.基于脑肠肽探讨功能性腹泻脾虚证“脾失健运”生物学机制及中药疗效机制[D].北京:北京中医药大学,2018.
张旭飞,蒋志滨,高洁,等.基于5-HT信号系统探讨痛泻要方治疗肝郁脾虚型溃疡性结肠炎的作用机制[J].中医学报,2021,36(10):2116-2121.
雷慧珺,曹建中,李亮,等.基于唾液转录组学探讨慢性胃炎和胃癌脾虚证的分子机制[J].世界中医药,2021,16(14):2196-2199.
HWANG M,KIM J N,KIM B J.Hesperidin depolarizes the pacemaker potentials through 5-HT(4) receptor in murine small intestinal interstitial cells of Cajal[J].Anim Cells Syst (Seoul),2020,24(2):84-90.
马潇,康安,徐珊,等.麸炒苍术不同提取物对小鼠脾虚泄泻的影响[J].南京中医药大学学报,2018,34(3):292-297.
马金鑫.参苓白术散对功能性腹泻脾虚证模型大鼠结肠上皮转运蛋白的影响[D].北京:中国中医科学院,2020.
韩晓春,吕佳楗,顾良臻,等.脾虚湿阻证大鼠肠道离子转运功能差异及薏苡仁作用[J].北京中医药大学学报,2020,43(7):583-591.
黄娟,施家希,施旭光,等.补中益气汤含药血清对caco-2细胞SGLT1、p38 MAPK通路及NHE3表达影响的研究[J].中药新药与临床药理,2018,29(6):707-712.
MATSUSAKA Y,CHEN X,ARIAS-LOZA P,et al.In vivo functional assessment of sodium-glucose cotransporters (SGLTs) using [(18)F]Me4FDG PET in rats[J].Mol Imaging,2022,2022:4635171.
吕林,王凤云,唐旭东,等.脾虚1号方对脾虚证功能性消化不良大鼠小肠葡萄糖转运蛋白1的影响[J].北京中医药大学学报,2017,40(6):456-462.
LIU P,JIANG L,KONG W,et al.PXR activation impairs hepatic glucose metabolism partly via inhibiting the HNF4α-GLUT2 pathway[J].Acta Pharm Sin B,2022,12(5):2391-2405.
任琦.对药茯苓-泽泻干预血脂异常痰浊证大鼠AMPK通路和肠道菌群的机制研究[D].广州:广州中医药大学,2021.
刘晓昱,舒畅.四君子汤对脾虚模型大鼠肽转运载体1转运功能及其基因表达的影响[J].浙江中医杂志,2020,55(11):836-838.
曹佳男.隔药饼灸对腹泻型肠易激综合征大鼠结肠5-HT、5-HT3R、5-HT4R表达的影响[D].长沙:湖南中医药大学,2019.
GU Y,QIN X,ZHOU G,et al.Lactobacillus rhamnosus GG supernatant promotes intestinal mucin production through regulating 5-HT4R and gut microbiota[J].Food Funct,2022,13(23):12144-12155.
于艳,贾天柱,才谦,等.茅苍术生品及麸炒品挥发油对脂多糖诱导人结肠上皮细胞炎症损伤的影响[J].中华中医药杂志,2022,37(3):1374-1379.
LUO L,CAI J,ZHOU Z,et al.Polysaccharides from Rhizoma Atractylodis Macrocephalae: A review on their extraction, purification, structure, and bioactivities[J].Evid Based Complement Alternat Med,2022,2022:2338533.
林雄,瞿领航,许静,等.苍术挥发油和苍术醇提物对溃疡性结肠炎模型小鼠的改善作用及其效果比较[J].吉林大学学报:医学版,2022,48(5):1247-1255.
0
Views
23
下载量
3
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution