文献类型：期刊文献

中文题名：基于IGF-1/PI3K/Akt信号通路探讨红芪多糖对糖尿病胃轻瘫大鼠胃窦平滑肌细胞凋亡的影响及作用机制

英文题名：Hedysari Radix Polysaccharide Regulates Apoptosis of Smooth Muscle Cells in Gastric Antrum of Rat Model of Diabetic Gastroparesis via IGF-1/PI3K/Akt Signaling Pathway

作者：苗琳琳[1,2];万生芳[1];张磊[1,3];李荣科[1];魏昭辉[1];朱小利[1,2];安惠[1,2]

第一作者：苗琳琳

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃省中医方药挖掘与创新转化重点实验室,兰州730030;[3]甘肃中医药大学敦煌医学与转化教育部重点实验室,兰州730030

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2024

卷号：30

期号：1

起止页码：130

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

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)。

语种：中文

中文关键词：糖尿病胃轻瘫;红芪多糖;胰岛素样生长因子-1(IGF-1)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路;胃窦平滑肌;凋亡;大鼠

外文关键词：diabetic gastroparesis;Hedysari Radix polysaccharide;insulin-like growth factor-1(IGF-1)/phosphatidylinositol-3 kinase(PI3K)/serine-threonine kinase(Akt)signaling pathway;smooth muscle of gastric antrum;apoptosis;rat

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/PI3K蛋白表达,以及红芪多糖低剂量组IGF-1、p-Akt/Akt、Bcl-2蛋白表达降低,Bax蛋白表达升高(P<0.05,P<0.01);与红芪多糖高剂量组比较,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、中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,P<0.01),IGF-1、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白表达降低,Bax蛋白表达升高(P<0.05,P<0.01);与红芪多糖中剂量比较,HPS低剂量组给药6、8周后体质量、大鼠胃排空率降低及胃窦组织平滑肌细胞凋亡指数升高(P<0.05,P<0.01),IGF-1、p-Akt/Akt、Bcl-2蛋白表达降低(P<0.05,P<0.01)。结论:红芪多糖能够在一定程度上保护胃窦平滑肌细胞凋亡损伤并促进胃动力,其机制可能与激活IGF-1/PI3K/Akt信号通路,上调IGF-1、p-PI3K、p-Akt、Bcl-2蛋白表达,下调Bax蛋白表达有关。
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).Compared with the model group,the 8 weeks of drug administration lowered the random blood glucose,increased the body mass and gastric emptying rate(P<0.05,P<0.01),decreased the apoptotic index of smooth muscle cells in the gastric antrum(P<0.05,P<0.01),up-regulated the protein levels of IGF-1,p-PI3K/PI3K,p-Akt/Akt,and Bcl-2,and down-regulated the protein level of Bax(P<0.05,P<0.01).Compared with the mosapride citrate group,the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass(P<0.05,P<0.01),low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased(P<0.05).The protein levels of IGF-1,p-PI3K/PI3K,p-Akt/Akt and Bcl-2(low dose)were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide(P<0.05,P<0.01).Compared with high-dose Hedysari Radix polysaccharide,low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration(P<0.05,P<0.01),and the low and medium doses decreased the gastric emptying rate,increased the apoptotic index of smooth muscle cells in the gastric antrum(P<0.05,P<0.01),down-regulated the protein levels of IGF-1,p-PI3K/PI3K,p-Akt/Akt,and Bcl-2,and up-regulated the protein level of Bax(P<0.05,P<0.01).Compared with the medium-dose group,the low-dose group of Hedysari Radix polysaccharide had lower body mass,lower gastric emptying rate in rats,higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration(P<0.05,P<0.01),and lower protein expression of IGF-1,p-PI3K/PI3K,p-Akt/Akt.Conclusion:Hedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway, as manifested by the up-regulated expression of IGF-1, p-PI3K, p-Akt, and Bcl-2 and down-regulated expression of Bax.