摘要
目的观察复方石斛合剂对糖尿病大鼠肝GCGR/PKA信号通路分子基因和蛋白表达的影响,探讨其抑制肝糖异生的分子机制。方法SPF级Wistar大鼠50只,随机选取11只作为正常组,其余39只高脂高糖饲料喂养6周后腹腔注射2次STZ,间隔72 h,筛选出糖尿病成模大鼠36只,随机分为模型组、二甲双胍组和石斛合剂组。石斛合剂组给予石斛合剂(11.3 g·kg^(-1))灌胃,二甲双胍组以二甲双胍灌胃(100 mg·kg^(-1)),正常组和模型组以生理盐水灌胃,连续给药12周。检测各组大鼠空腹血糖(Fasting blood glucose,FBG);ELISA方法检测血清胰高血糖素(Glucagon);RT-PCR法检测GCGR/PKA信号通路的肝糖异生相关基因表达。免疫组化法和Western Blot法检测上述信号通路肝糖异生相关蛋白表达。结果石斛合剂组FBG、Glucagon较模型组显著下降(P<0.01);免疫组化结果显示石斛合剂能显著抑制GCGR、G6Pase的表达(P<0.01);RT-PCR和Western blot结果显示石斛合剂组β-catenin、TCF7L2基因和蛋白表达增加(P<0.05,P<0.01);GCGR、PKA基因和蛋白表达减少(P<0.05,P<0.01)。结论石斛合剂能控制糖尿病模型大鼠的空腹血糖,抑制肝糖异生,可能与其通过调控GCGR/PKA信号通路相关信号分子的基因和蛋白表达有关。
Objective To observe the effect of compound Shihu mixture on the expression of molecular genes and proteins of the liver GCGR/PKA signaling pathway in diabetic rats,and to explore its molecular mechanism of inhibiting liver gluconeogenesis.Methods 50 SPF-grade healthy Wistar rats,11 were randomly selected as the normal group,and the remaining 39 rats were fed with high-fat and high-sugar diet for 6 weeks and then injected with STZ,interval 72 h 36 diabetic model rats were screened out,and then they were randomly divided into 3 groups(model group,Shihu mixture group and metformin group).The Shihu mixture group was given Shihu Mixture(11.3 g·kg^(-1))by gavage,and the metformin group was given Metformin by gavage(100 mg·kg^(-1)).The model group and the normal group were given saline intragastrically for 12 weeks.All animals in each group were collected 24 hours after the last medication,and the fasting blood glucose(FBG)was detected.The serum glucagon(Glucagon)was detected by ELISA.RT-PCR method to detect the expression of hepatic gluconeogenesis-related genes in the GCGR/PKA signaling pathway.Immunohistochemical method and Western blot method were used to detect the expression of hepatic gluconeogenesis-related proteins in the above signal pathways.Results Compared with the model group,FBG and Glucagon in the Shihu mixture group significantly decreased(P<0.01);the immunohistochemical results showed that the Shihu mixture could inhibit the expression of GCGR and G6 Pase(P<0.01);RT-PCR and Western-blot results showed that the protein and mRNA expressions of β-catenin and TCF7 L2 in the Shihu mixture group increased(P<0.05,P<0.01);the protein and mRNA expressions of GCGR and PKA decreased(P<0.05,P<0.01).Conclusion Shihu mixture can effectively reduce fasting blood glucose in diabetic rats and inhibit liver gluconeogenesis in diabetic model rats,which may be related to the regulation of the gene and protein expression of related signal molecules in the GCGR/PKA signaling pathway.
作者
林心君
刘佳绣
陈勇
何昱霖
施红
LIN Xin-jun;LIU Jia-xiu;CHEN Yong;HE Yu-lin;SHI Hong(Fujian University of Traditional Chinese Medicine,Fuzhou Fujian 350122,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2022年第5期1062-1065,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81503438,81973827)
福建省自然科学基金(2019J01332)。
