风药协同六味地黄汤治疗肾阴虚型糖尿病的机制OA

[Objective]To investigate the synergistic effect and mechanism of wind medicinals(Puerariae Lobatae Radix,Atractylodis Rhizoma and Rhizoma et Radix Notopterygii)combined with Liuwei Dihuang Decoction(LWDHD)in treating kidney Yin deficiency diabetes mellitus,based on the Xuanfu theory.[Methods]Seventy male SD rats were randomly divided into normal group(n=10)and modeling group(n=60).The normal group was fed with a standard maintenance diet,while the modeling group was fed a high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ)and subcutaneous injection of thyroxine to establish a rat model of kidney Yin deficiency diabetes.Fifty successfully modeled rats were then randomly allocated into model group,LWDHD group,wind medicinals group,combination group(wind medicinals+LWDHD group),and metformin group.The normal and model groups were administered an equal volume of 0.9%sodium chloride solution by gavage,the LWDHD group received LWDHD(crude drug 7.875 g·kg-1),the wind medicinals group received the wind medicinals decoction(crude drug 3.150 g·kg-1),the combination group received LWDHD combined with the wind medicinals decoction(crude drug 11.025 g·kg-1),and the metformin group received metformin solution(200 mg·kg-1),gavage once daily for 6 weeks.The general state and body weight changes of the rats were observed,and FBG was measured.Serum fasting insulin(FINS)levels were detected by ELISA,and the homeostatic model assessment of insulin resistance(HOMA-IR)was calculated.HE staining was used to observe pathological changes in the pancreas.Immunohistochemistry was performed to detect insulin(INS)expression in pancreatic tissue.The protein expression levels of phosphoinositide 3-kinase(PI3K),phosphorylated-PI3K(p-PI3K),protein kinase B(Akt)and phosphorylated-Akt(p-Akt)in pancreatic tissue were detected with Western blot.Real-time PCR was performed to detect the mRNA expression levels of insulin receptor substrate-1(IRS-1)and glucose transporter 4(GLUT4)in pancreatic tissue.[Results]Compared with normal group,model group showed mental fatigue,dull fur,increased polydipsia,polyphagia,polyuria,reduced activity,and significantly decreased body weight in the later stage(P<0.01);serum FINS,FBG and HOMA-IR were significantly elevated(P<0.05).The pancreatic islets showed significant atrophy,irregular morphology,reduced islet cell count,along with vacuolar degeneration and inflammatory cell infiltration.The protein expression of INS,p-PI3K/PI3K and p-Akt/Akt,as well as the mRNA expression of IRS-1 and GLUT4 in pancreatic tissue were significantly decreased(P<0.05).After six weeks of drug intervention,compared with model group,the general condition of rats in all drug-administered groups was improved to varying degrees,with combination group showing the most significant improvement in general state and islet morphological structure repair.The combination group,LWDHD group and metformin group exhibited significant reductions in serum FINS,FBG and HOMA-IR,along with significant increases in pancreatic tissue expression of INS,p-PI3K/PI3K and p-Akt/Akt proteins,as well as IRS-1 and GLUT4 mRNA levels(P<0.05).In the wind medicinals group,the expression of INS and p-PI3K/PI3K protein in the pancreatic tissue were significantly elevated(P<0.05).Compared with combination group,both LWDHD group and wind medicinals group showed significantly inferior effects in reducing serum FINS,FBG and HOMA-IR,and in enhancing pancreatic tissue expression of INS,p-PI3K/PI3K,p-Akt/Akt proteins,as well as IRS-1 and GLUT4 mRNA levels(P<0.05).[Conclusion]Wind medicinals can enhance the hypoglycemic effect of LWDHD,and its mechanism may be related to"opening the Xuanfu",enhancing the Yin-nourishing and kidney-tonifying effects of LWDHD,improving pancreatic microcirculation,and synergistically activating the PI3K/Akt/GLUT4 signaling pathway.