摘要
目的通过体内外实验探究哈尼族验方七花颗粒对2型糖尿病糖脂代谢的改善作用。方法取大鼠肝脏间质细胞(BRL-3A)接种于六孔板中,分为空白组、棕榈酸组、非诺贝特组、七花颗粒含药血清5.4、10.8、21.6 g/kg组;除空白组外,其余各组以0.2 mmol/L棕榈酸(Palmitic acid,PA)干预24 h,加入含药血清,再持续孵育24 h。测定各组BRL-3A细胞的增殖率;检测各组细胞上清培养液中总胆固醇(Total cholesterol,T-CHO)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(High-density lipoprotein cholesterol,HDL-C)的含量;吸弃细胞培养液,检测细胞裂解液中甘油三酯(Triglyceride,TG)含量;油红染色,检测细胞中脂质含量。同时,取45只大鼠分为空白组、模型组、非诺贝特组(0.225 g/kg)、七花颗粒复方5.4 g/kg组、七花颗粒复方10.8 g/kg组,空白组给予普通饲料喂养,其余各组给予高脂饲料喂养42 d。第43 d开始,除空白组,各组一次性腹腔注射0.25%链脲佐菌素(Streptozotocin,STZ)溶液,同时,灌胃给予相应药物,持续14 d。测定大鼠体重增长量和肝脏指数;检测血清中空腹血糖(Fastingbloodglucose,FBG)、空腹胰岛素(Fastinginsulin,FINS)含量,计算胰岛素抵抗指数(Insulin resistance index,ISI),同时检测血清游离脂肪酸(Free fatty acid,FFA)及肝组织中肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)的含量;HE染色观察胰腺组织的病理变化;免疫组化检测胰岛α、β细胞表达。结果与PA组比较,七花颗粒含药血清10.8、21.6 g/kg组BRL-3A细胞增殖率显著升高(P<0.01);七花颗粒复方含药血清5.4、21.6 g/kg组T-CHO、LDL-C、TG含量显著降低(P<0.05),HDL-C的含量显著升高(P<0.05);油红染色表明,七花颗粒复方含药血清5.4、10.8、21.6 g/kg组细胞质中的脂质明显减少。动物实验结果显示,与模型组相比,七花颗粒复方10.8 g/kg组体重显著增长(P<0.05);七花颗粒复方5.4 g/kg组肝指数显著下降(P<0.05);七花颗粒复方5.4、10.8 g/kg组血清中FBG、FINS、FFA和ISI均显著降低(P<0.05或P<0.01);同时,TNF-α的含量显著降低(P<0.01)。HE染色结果显示,七花颗粒复方5.4、10.8 g/kg组大鼠胰导管及血管周围少量淋巴细胞散在分布,局部血管扩张,胰岛细胞数量及面积明显增多;免疫组化结果显示,七花颗粒复方5.4、10.8 g/kg组胰岛α细胞面积明显减少,胰岛β细胞面积明显增多。结论七花颗粒复方改善T2DM糖脂代谢,可能是通过改善胰岛细胞的功能,增加胰岛素对血糖的敏感性,改善胰岛素抵抗,减少胰岛素和胰高血糖素的分泌,从而降低空腹血糖水平;同时通过降低TNF-α含量,抑制体内脂肪分解,促进脂肪细胞对FFA的摄取,进一步降低FFA,从而调节TG、T-CHO、HDL-C及LDL-C水平,改善糖脂代谢异常,缓解T2DM。
Objective To investigate the ameliorative effect of Qihuakeli,a Hani formula,on glycolipid metabolism in type 2 diabetes mellitus by in vivo and in vitro experiments.Methods Rat liver mesenchymal stromal cells(BRL-3A)were inoculated in six-well plates and divided into blank,palmitic acid,fenofibrate,and Qihuakeli serum-containing 5.4,10.8,and 21.6 g/kg groups.Except for the blank group,the remaining groups were intervened with 0.2 mmol/L palmitic acid(PA)for 24 hour,and then added with drug-containing serum,and then continued to incubate for 24 hour.The proliferation rate of BRL-3A cells in each group was determined.Total cholesterol(T-CHO),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)concentrations in the supernatant of each cell group were measured,cell culture medium was aspirated and discarded,triglyceride(TG)concentration in the cell lysate.The lipid content of the cells was determined by measuring and staining with red oil.Meanwhile,45 rats were taken and divided into blank group,model group,fenofibrate group(0.225 g/kg),Qihuakeli compound 5.4 g/kg group,and Qihuakeli compound 10.8 g/kg group,the blank group was given normal feed and the rest of the groups were given high-fat feed for 42 day.Beginning on the 43rd day,each group,except the blank group,was injected with a single intraperitoneal injection of Starting from the 43rd day,except the blank group,each group was given a one-time intraperitoneal injection of 0.25%streptozotocin(STZ)solution,and at the same time,the corresponding drugs were given by gavage for 14 day.The rats'weight gain and liver index were measured.Serum fasting blood glucose(FBG)and fasting insulin(FINS)were detected,and the insulin resistance index(ISI)was calculated.Serum free fatty acid(FFA)levels and tumor necrosis factor-α(TNF-α)in liver tissue were also detected.HE staining was used to detect pathological changes in the pancreas.Pathological changes were observed in the tissues,and isletαandβcell expression was detected by immunohistochemistry.Results Compared to the PA group,the accumulation rate of BRL-3A cells was significantly higher(P<0.01)in the 10.8 and 21.6 g/kg Qihuakeli-containing serum groups.The levels of T-CHO,LDL-C and TG in the 5.4 and 21.6 g/kg serum groups were significantly lower(P<0.05),and HDL-C levels significantly increased(P<0.05).Oil red staining results showed that lipids in the cytoplasm of the 5.4,10.8 and 21.6 g/kg.Qihuacel-containing groups significantly reduced.Compared to the model group,the body weight of the 10.8 g/kg group containing Qihuakeli granules increased significantly(P<0.05).The liver index of the 5.4 g/kg group containing Qihuakeli decreased significantly(P<0.05).The serum indices of FBG,FINS,FFA and insulin resistance of the 5 g/kg group containing Qihuakeli decreased significantly(P<0.05).In the 5.4,10.8 g/kg groups,all serum FBG,FINS,FFA and insulin resistance indices significantly reduced in the 5.4 and 10.8 g/kg Qihuakeli groups(P<0.05 or P<0.01).TNF-αlevels were significantly reduced(P<0.01).HE staining showed that a small number of lymphocytes were scattered in the pancreatic ducts and perivascular area of the rats in the Qihuakeli 5.4 and 10.8 g/kg groups,the local vasodilatation was observed,the number of pancreatic islet cells and the area of islet cells significantly increased.Immunohistochemical study was further used.The results of immunohistochemistry showed that the area of pancreatic isletα-cells significantly reduced and the area of pancreatic isletβ-cells significantly increased in Qihuakeli 5.4 and 10.8 g/kg groups.Conclusion Qihuakeli compound improved glucose-lipid metabolism in T2DM,probably by improving the function of pancreatic islet cells,increasing the sensitivity of insulin to blood glucose,improving insulin resistance,decreasing the secretion of insulin and glucagon,and thus lowering the level of fasting blood glucose.Meanwhile,by decreasing the content of TNF-α,inhibiting lipolysis in the body,and promoting the uptake of FFA by adipocytes,and further lowering the FFA.Thus,it regulates the levels of TG,T-CHO,HDL-C and LDL-C,improves the abnormalities of glucose and lipid metabolism,and alleviates T2DM.
作者
万艳
吴海妹
刘非凡
白元美
张林翱
罗世芳
吴雪
解宇环
孔春芹
郭沛鑫
WAN Yan;WU Haimei;LIU Feifan;BAI Yuanmei;ZHANG Lin'ao;LUO Shifang;WU Xue;XIE Yuhuan;KONG Chunqin;GUO Peixin(School of Ethnomedicine,Yunnan University of Traditional Chinese Medicine,Kunming 650500,China;School of Traditional Chinese Medicine,Yunnan University of Traditional Chinese Medicine,Kunming 650500,China;School of Basic Medical Sciences,Yunnan University of Traditional Chinese Medicine,Kunming 650500,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第5期1386-1398,共13页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
云南省科技厅基础研究计划项目(2018FF001(-075)):民族药七花颗粒对胰岛素抵抗模型大鼠的干预作用及作用机制研究,负责人:孔春芹
云南省科技厅计划项目(202105AG070014):云南省配方颗粒重点实验室,负责人:陈钟
国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023192):少数民族药学(傣药学),负责人:冯德强。
关键词
2型糖尿病糖脂代谢
七花颗粒
大鼠肝脏间质细胞
作用机制
Glycolipid metabolism in type 2 diabetes mellitus
Seven-flower granules
Rat liver mesenchymal stromal cells
Mechanism of action
