| Backgrounds:The prevalence of type 2 diabetes mellitus(T2DM)is increasing year by year,which has become a major global public health problem.Islet β cell dysfunction is considered to be a key link in the pathogenesis of T2DM.Recent studies have found that the accumulation of senescent β cells in pancreatic tissues is an important factor leading to β cell dysfunction.Middle-aged and elderly male T2DM patients often show symptoms of kidney-yang deficiency in clinical practice.Studies have shown that traditional Chinese medicine plays an important role in the prevention and treatment of T2DM and its complications.Some traditional Chinese medicine with the effect of tonifying kidney and strengthening Yang can play the role of anti-aging and regulating glucolipid metabolism.The clinical application of Duzhong Congrong decoction in the treatment of male T2DM patients with kidney-yang deficiency has achieved initial results.Duzhong Congrong decoction may improve the function of β cells by delaying cellular senescence,which is beneficial to glycemic control.Objectives:To observe the effects of Duzhong Congrong decoction on blood glucose and testosterone levels in male T2DM patients.To clarify the effects of Duzhong Congrong decoction on pancreatic β cells and testicular function in diabetic mice,and to investigate whether Duzhong Congrong decoction can delay the senescence of β cells.Methods:(1)Based on the propensity score matching,male T2DM patients who attended the department of endocrinology in Jiangsu Province Hospital on Integrated Chinese and Western Medicine from September 2021 to December 2022 were retrospectively analyzed,with those who had taken Duzhong Congrong decoction as the intervention group and those who did not take Duzhong Congrong decoction as the control group.The improvement of fasting blood glucose,glycosylated hemoglobin,body mass index,blood pressure,blood lipids,testosterone levels and symptoms of kidney Yang deficiency in male T2DM patients were observed after taking Duzhong Congrong decoction for 3 months.(2)Six-month-old C57BL/6 mice were selected and divided into normal chow diet group(NC)and high-fat diet group.The mice in the high-fat diet group were fed with 60%high-fat diet to construct a diabetic model,and then the successfully modeled diabetic mice were randomly divided into five groups:T2DM model group(Mod),low-dose Duzhong Congrong decoction group(DZCRL),high-dose Duzhong Congrong decoction group(DZCRH),WuZi Yanzong pill group(WZYZ),and metformin group(Met).The mice in the DZCRL and DZCRH groups were gavaged with low and high doses of Duzhong Congrong decoction,while the WZYZ,Met groups were gavaged with Wuzi Yanzong pill solution and metformin solution,Mod and NC groups were gavaged with ultra-clean water,once a day for 8 weeks.After 8 weeks of intervention,the body weight,fasting glucose,glucose tolerance,homeostatic model assessment of β(HOMA-β)and serum testosterone levels were measured in each group.The morphology of pancreas and testis in each group was observed.The sperm concentration,motility and malformation of mice were recorded.The apoptosis and β-galactosidase(β-gal)activity levels of pancreas were assessed.The expression of insulin genes and genes related to cellular senescence were observed in each group of mice.Results:(1)After 3 months of treatment,the fasting blood glucose level of Duzhong Congrong decoction group was lower than that in the control group(P<0.05),and the decrease in FPG was greater than that in the control group(P<0.05).There was no significant difference in testosterone levels in the intervention group compared with the control group(P<0.05),but their improvement from baseline was greater than that in the control group(P<0.05).The levels of body mass index,systolic blood pressure,diastolic blood pressure,glycosylated hemoglobin,total cholesterol,triglycerides,LDL cholesterol,HDL cholesterol,luteinizing hormone and follicle stimulating hormone were not significantly different between the two groups.The symptom scores of both the intervention and control groups were significantly improved compared with baseline.However,the intervention group had a greater reduction in symptom scores.(2)Diabetic mice were successfully modeled after 14 weeks of high-fat diet feeding.After 8 weeks of intervention,FPG levels were significantly lower in the DZCRL and Met groups compared with the Mod group(P<0.05).The intraperitoneal glucose tolerance test showed that the area under the blood glucose curve in the DZCRL,WZYZ and Met groups was lower than that in the Mod group(P<0.05).No significant difference was observed in fasting insulin levels between the groups detected by ELISA,while the calculated HOMA-β level in the DZCRL group was significantly higher than that in the Mod group(P<0.05).HE staining showed that the islets in the NC group were morphologically.The islets in the Mod group had irregular morphology and increased cross-sectional area,while those in the DZCRL,WZYZ and Met groups had different degrees of improvement compared with the Mod group,and those in the DZCRH group had irregular morphology.The epithelium of the germinal tubules in the testis tissue of the Mod group was damaged,the structure was loose,the germinal cells were not well arranged,and some of them were detached and necrotic.The testicular morphology of the DZCRL and WZYZ groups were significantly improved compared with the Mod group and were similar to that of the NC group.The testicular morphology of the DZCRH and Met groups was slightly improved compared with that of the Mod group.Sperm density and motility decreased and malformation rate increased in the Mod group compared to the NC group.The DZCRL group showed significant improvement in sperm concentration,motility and malformation rate compared to the Mod group.After 8 weeks of drug intervention,no significant differences were observed in testosterone levels as well as pancreatic apoptosis in the mice of each group.The results of pancreatic β-gal staining showed that the area of positive β-gal staining was significantly increased in the Mod group compared with the NC group,while the area of positiveβ-gal staining was decreased in the DZCRL,WZYZ and Met groups compared with the Mod group.Fluorescence quantitative PCR results showed that the expression of Ins1 and Ins2 were significantly decreased in the Mod group compared with the NC group,and the expression of Insl and Ins2 was significantly elevated in the DZCRL and WZYZ groups compared with the Mod group(P<0.001).The expression of p16,p21,p53,and Rb were significantly increased in the Mod group compared with the NC group,and DZCRL reversed these expressions.The expression of these genes was also decreased to different degrees in the WZYZ and Met groups compared to the Mod group.Conclusions:(1)Duzhong Congrong decoction is beneficial in improving fasting glucose and increasing testosterone levels in men with type 2 diabetes based on conventional treatment.(2)Duzhong Congrong decoction can decrease the fasting blood glucose level,improve the function of β cells in diabetic mice,and protect the spermatogenic cells in diabetic mice.Duzhong Congrong decoction may delay islet cell senescence by decreasing the expression of islet cell senescence-related genes,which in turn improves islet β cell function. |
PDF Full Text Request