摘要
目的:观察红芪多糖对糖尿病胃轻瘫(DGP)大鼠胃窦平滑肌细胞凋亡及胰岛素样生长因子-1/磷脂酰肌醇3-激酶/蛋白激酶B(IGF-1/PI3K/Akt)通路蛋白表达的影响,探讨其作用机制。方法:将62只Wistar雄性大鼠随机分为空白组12只、造模组50只。造模组采用小剂量多次腹腔注射链脲佐菌素联合不规律高脂高糖饮食法4周构建大鼠DGP模型。将成模大鼠随机分为模型组、莫沙必利组和红芪多糖高、中、低剂量组。红芪多糖高、中、低剂量组分别予红芪多糖200、100、50 mg·kg^(-1)灌胃,莫沙必利组予枸橼酸莫沙必利1.35 mg·kg^(-1)灌胃,空白组和模型组予等体积纯净水灌胃,每日1次,连续8周。每2周测定大鼠随机血糖及体质量,检测胃排空率;苏木素-伊红(HE)染色观察胃窦组织平滑肌病理形态;原位末端标记法(TUNEL)染色检测胃窦组织平滑肌细胞凋亡指数;免疫组化法检测胃窦组织平滑肌IGF-1、磷酸化(p)-PI3K、p-Akt蛋白的表达;蛋白免疫印迹法(Western blot)检测胃窦组织平滑肌中IGF-1、p-PI3K/PI3K、p-Akt/Akt、B细胞淋巴瘤-2(Bcl-2)及Bcl-2相关X蛋白(Bax)的蛋白表达水平。结果:与空白组比较,模型组大鼠各时间点随机血糖显著升高(P<0.01),体质量、胃排空率显著降低(P<0.01),胃窦组织平滑肌细胞凋亡指数显著升高(P<0.01),IGF-1、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白表达降低,Bax蛋白表达显著升高(P<0.01);与模型组比较,各给药组大鼠给药8周后随机血糖降低,体质量、胃排空率均明显升高(P<0.05,P<0.01)、胃窦组织平滑肌细胞凋亡指数明显降低(P<0.05,P<0.01),IGF-1、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白表达升高,Bax蛋白表达明显降低(P<0.05,P<0.01);与莫沙比利组比较,红芪多糖低剂量组给药6、8周后随机血糖升高、体质量减轻(P<0.05,P<0.01),红芪多糖低剂量组大鼠胃排空率降低、胃窦组织平滑肌细胞凋亡指数升高(P<0.05),红芪多糖各剂量组p-PI3K/P
Objective:To observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1(IGF-1)/phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt)pathway in the rat model of diabetic gastroparesis(DGP).Method:Sixty-two Wistar male rats were randomized into a blank group(n=12)and a modelling group(n=50).The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks.The modeled rats were randomized into model group,mosapride citrate(1.35 mg·kg^(-1)),and high-,medium-,and low-dose(200,100,and 50 mg·kg^(-1),respectively)Hedysari Radix polysaccharide groups.The rats were administrated with corresponding drugs by gavage,and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks.The random blood glucose and body mass were measured every 2 weeks,and gastric emptying rate was calculated.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of smooth muscle in gastric antrum,and terminal deoxynucleoitidyl transferase-mediated nick-end labeling(TUNEL)was employed to detect the apoptosis of smooth muscle cells in the gastric antrum.The expression of IGF-1,phosphorylated(p)-PI3K,and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry.Western blot was employed to determine the protein levels of IGF-1,p-PI3K/PI3K,p-Akt/Akt,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in the smooth muscle of the gastric antrum.Result:Compared with the blank group,the model group showed elevated random blood glucose at all time points(P<0.01),decreased body mass and gastric emptying rate(P<0.01),increased apoptotic index of smooth muscle cells in the gastric antrum(P<0.01),down-regulated protein levels of IGF-1,p-PI3K/PI3K,p-Akt/Akt,and Bcl-2,and up-regulated protein level of Bax(P<0.01).Com
作者
苗琳琳
万生芳
张磊
李荣科
魏昭辉
朱小利
安惠
MIAO Linlin;WAN Shengfang;ZHANG Lei;LI Rongke;WEI Zhaohui;ZHU Xiaoli;AN Hui(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730030,China;Gansu Key Laboratory of Traditional Chinese Medicine Formula Innovation and Transformation,Lanzhou 730030,China;Key Laboratory of Dunhuang Medicine,Ministry of Education,Gansu University of Chinese Medicine,Lanzhou 730030,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第1期130-139,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金重大项目(82060914)
甘肃省中医药研究中心课题(zyzx-2020-zx11)。
