| Purpose:1Through in vitro experiment,to explore whether the anti-aging effect of Huangqi Compound Granules on renal mesangial cells induced by high sugar is related to its effect of delaying mitochondrial damage and anti-oxidative stress.2By observing the clinical effect of Huangqi Compound granules in the treatment of early diabetic nephropathy(spleen-kidney deficiency combined with blood stasis type),the therapeutic effect of Huangqi compound granules in the treatment of early diabetic nephropathy(spleen-kidney deficiency combined with blood stasis type)and the clinical mechanism of reducing albuminuria are analyzed,in order to provide treatment methods and ideas for the treatment of early diabetic nephropathy and reducing albuminuria type.Materials and methods:1Drug serum was prepared by SPF grade Wistar male rats aged 8 weeks.Rat mesangial cells(RMC)were divided into 5groups.Namely,control group(5.5mmol/L glucose + 20% normal serum group),model group(25mmol/L glucose + 20% normal serum group),low concentration group(25mmol/L glucose + 5% drug serum + 15% normal serum group),medium concentration group(25mmol/L glucose + 10% drug serum + 10% normal serum group)Group)and high concentration group(25mmol/L glucose + 20% drug serum)were detected by cell proliferation and toxicity detection reagent(Cell Counting Kit-8,CCK8).The effects of high sugar and drug-containing serum on mitochondrial membrane potential and mitochondrial damage of RMC were detected by JC-1 staining.The average fluorescence intensity of reactive oxygen species(ROS)was detected by multifunctional enzyme marker to observe the effect of high glucose and drug-containing serum on oxidative stress of RMC.The effect of high sugar and drug-containing serum on RMC senescence was observed by detecting the number of β-gal staining positive cells.2 Outpatient cases from the general Internal medicine Department of Jiahe Hospital of Traditional Chinese Medicine of Liaoning were selected.All the collected cases should meet the inclusion criteria.A total of 50 outpatient cases were collected and randomly divided into control group and treatment group at a ratio of 1:1.The control group was given conventional western medicine treatment for diabetic nephropathy,while the treatment group was given oral administration of Huangqi Compound Granules based on conventional western medicine treatment.After 8 weeks of observation,the medium symptom scores,blood glucose,UACR and other related laboratory indexes of the two groups before and after treatment were compared.Data were collected and analyzed by SPSS25.0.To evaluate the clinical effect of Huangqi Compound Granule on early diabetic nephropathy(spleen-kidney deficiency combined with blood stasis type)and its influence on urinary protein and blood glucose.Results:1The morphological changes of rat mesangial cells were induced by high glucose.The cell viability decreased at 24 h and 72 h,and the cell proliferation was abnormal at 48 h.2High glucose stimulation can make RMC mitochondria produce excessive ROS,and the maximum value is reached at 48 h.The ROS production level of cells can further reflect the degree of oxidative stress.Therefore,this experiment can indicate that high glucose environment can induce the oxidative stress response of RMC,and the degree of oxidative stress is the most obvious at 48 h.3High glucose stimulation can damage the structure and function of mitochondria by down-regulating the mitochondrial membrane potential level,and make RMC produce more ROS under high glucose environment,thus inducing the aging of RMC.Therefore,the results of this experiment show that high sugar can reduce the membrane potential level of mitochondria and destroy the structure and function,and the mitochondrial damage degree is the most significant at 48 h,so as to further strengthen the level of oxidative stress response induced by high sugar and induce senescence of RMC,and the senescence degree is the most obvious at 48 h.4The serum of Huangqi Compound Granules can inhibit the production of mitochondrial ROS in RMC under high glucose environment,so as to exert its antioxidant stress effect,and the antioxidant stress effect is most obvious in the medium concentration group at 48 h.5The serum of Huangqi Compound Granules can improve the mitochondrial damage by up-regulating the membrane potential of mitochondria and delay the oxidative stress level of RMC in the high glucose environment,so as to delay the aging process of RMC induced by high sugar.The maximum effect can be achieved by delaying the oxidative stress response and aging process of RMC in the medium concentration of the serum at 48 h.6Huangqi Compound Granules can significantly improve the TCM symptoms of patients;Influence on blood glucose level of patients: although it can significantly reduce blood glucose,compared with the control group,it has no significant advantage in reducing FPG and 2h PG;Effect on patients with albuminuria: UACR can be significantly reduced in patients,and compared with the control group,there is absolute advantage.Therefore,Huangqi Compound Granules have certain advantages in improving patients’ TCM symptoms and reducing UACR.Conclusions:1The morphological changes of rat mesangial cells were induced by high glucose.The cell viability decreased at 24 h and 72 h,and the cell proliferation was abnormal at 48 h.2High sugar can induce oxidative stress reaction of RMC and produce a large number of ROS,and the oxidative stress reaction is most obvious at 48 h.3Huangqi Compound Granule medium concentration group showed the most significant anti-oxidative stress effect after 48 h intervention.4Huangqi Compound Granule medium concentration group has the most significant effect on delaying aging after 48 h intervention.5High sugar can induce the decrease of mitochondrial membrane potential,while the intervention of Huangqi Compound granule medium concentration group for48 h can significantly increase the mitochondrial membrane potential.6Huangqi Compound Granules in the treatment of early diabetic nephropathy(spleen-kidney deficiency combined with blood stasis type)can significantly improve the symptoms of traditional Chinese medicine,reduce the UACR of patients,reduce albuminuria,and effectively delay the progression of the disease to end-stage renal disease. |
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