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The study of renal tissue in DN rats SREBP influence mechanism of Yiqi Jiedu activating method of early

Author: JinZuoTing
Tutor: LiJingLin
School: Liaoning University of Traditional Chinese Medicine
Course: Traditional Chinese Medicine
Keywords: diabetic nephropathy Yiqi Jiedu activating method SREBP-1c SREBP-2c
CLC: R277.5
Type: PhD thesis
Year: 2013
Downloads: 17
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Abstract


Purpose:Investigate the effects of Yiqi Jiedu meridians of Chinese medicine compound on early diabetic nephropathy (DN) rats pharmacodynamic indexes and renal tissue of sterol regulatory element binding protein (SREBP-lc, SREBP-2c) and the effect of gene and protein expression, observation of Yiqi Jiedu adjustable mechanism of traditional Chinese medicine compound on the protective effect and its role.Material and method:The SPF level of90healthy male Wistar rats, fed a week, were randomly divided into2groups:normal control group (10rats), model group (80rats). The control group was given normal diet, model group fed with high fat diet.1weeks Houzao group rats were given intraperitoneal injection of STZ42mg/kg, normal control group, were injected with citrate buffer. After72hours the rats were measured fasting blood glucose, identified as DM models over16.7mmol/L. According to the blood glucose level of each group were randomly divided into two groups, model group, prevention group, high dose group, low dose group, western medicine group. Effect of Yiqi Jiedu meridians of Chinese medicine compound selection of Chinese medicine decoction and granule dissolved in distilled water. The treatment group daily dosage of human in rats (g/kg) equivalent dose6.3times computation; dosage Chinese medicine high dose treatment group was3times of low dosage Chinese medicine treatment groups. The normal control group, model control group rats were given normal saline. Traditional Chinese medicine prevention group were given orally active compound Yiqi Jiedu immediately in the DM model,1times a day, a total of10weeks of administration of traditional Chinese medicine; low dose group, high dose treatment group and positive control group of Western medicine in DM model after2weeks, the DN model after the perfusion of Chinese herbal compound,1times a day, a total of8weeks of administration. Collection of urine of rats in the stomach after urinary albumin measured; abdominal aortic puncture blood blood glucose, glycosylated hemoglobin, serum β2microglobulin levels, blood lipid, renal function:creatinine blood urea nitrogen; HE in renal tissues of rats were observed under the light microscope, PAS staining images change by ELISA method; detection of serum DN SREBP-lc in rats, the content of SREBP-2c; the expression of SREBP-2c gene and protein SREBP-1c, observation of Yiqi Jiedu meridians of Chinese medicine compound on early DN in kidney of rat.Results:1. The changes of blood glucose of rats in each group:rats blood glucose changes showed, compared with the normal group, the model group rats blood glucose increased significantly, there was significant difference between the two groups (P<0.01); compared with the model group, drug intervention treatment of blood glucose of rats in each group were significantly decreased (P<0.01), but did not reach normal glucose range of values, in the use of drugs in the treatment group, Chinese medicine prevention group blood sugar decreased the most obvious, the experimental results show that:the use of drug intervention therapy can significantly reduce early DN rats blood glucose level increases, the best intervention effect of Chinese medicine prevention group, western medicine group and the effect of intervention time.2. Changes in rats of glycated hemoglobin:compared with the normal control group, the content of glycosylated hemoglobin in rats of model group increased obviously, there was significant difference between two groups (P<0.01); compared with the model group, glycosylated hemoglobin of rats in each treatment group decreased significantly (P<0.01), but compared with the normal group there was a significant difference (P<0.01), indicating that the drug intervention can effectively reduce the DN rat glycated hemoglobin, but has not yet reached the normal level; traditional Chinese medicine prevention intervention treatment group was most obvious in the drug intervention group.3. Urine albumin changes of rats in each group:compared with the normal group, urinary albumin in rats of model group increased obviously, there was significant difference between two groups (P<0.01); compared with the model group, albumin in the urine of rats in each group using the drug intervention group were significantly decreased (P<0.01), but with the the normal group were significant difference (P<0.01), indicating that the drug intervention can effectively reduce the content of albumin DN in rat urine, but has not yet reached the normal level; traditional Chinese medicine prevention intervention treatment group was most obvious in the treatment group, the experimental results show that, the most obvious effect of prevention and treatment group content of albumin in rat urine.4. Changes of2-microglobulin β groups of rats:compared with the normal group, the blood β2-microglobulin in the rats of model group increased obviously, there was significant difference between two groups (P<0.01); compared with the model group, the blood β2-microglobulin rats using drug intervention group were significantly decreased (P<0.01), but compared with the normal group, there was significant difference (P<0.01), indicating that the drug intervention can effectively reduce DN rats blood β2-microglobulin; the use of drug treatment groups had no significant difference (P>0.05). The experimental results show that, traditional Chinese medicine prevention treatment group was most obvious influence on the content of2-β2-microglob.5. Changes in serum creatinine of rats in each group:the serum creatinine content in rats of every group had no obvious change, no significant differences among the groups (P>0.05). The use of drug intervention groups, traditional Chinese medicine prevention group was the highest, compared with the western medicine group, they are no significant difference between the two groups (P>0.05).6. Changes of serum urea nitrogen of rats in each group:compared with the normal group, serum urea nitrogen in the rats of model group were significantly elevated, there was significant difference between two groups (P<0.01); compared with the model group, the serum drug use rats urea nitrogen content in Chinese medicine prevention intervention treatment group was most obvious in the use of drug intervention group (P<0.01), in the prompt intervention can effectively reduce the blood urea nitrogen content in serum of DN rats, but has not yet reached the normal level. The experimental results show that, traditional Chinese medicine prevention treatment group was most obvious effect on serum urea nitrogen.7. The pathological changes of kidney of ratsHE staining:observation at low magnification, rats in normal control group, renal tubular and glomerular mesangial matrix without obvious pathological changes; high magnification, model group, glomerulosclerosis, mesangial cells and mesangial matrix proliferation, vacuoles in renal tubular epithelial cell deformation; each treatment group and traditional Chinese medicine prevention group change degree renal pathology was obviously lighter than the model control group, traditional Chinese medicine prevention group rat kidney glomerular and tubular than normal rats increased slightly; very high dose and the western medicine group rat kidney glomerular volume size, but were higher than normal group.PAS staining:normal rats, renal tubular and glomerular mesangial matrix without obvious pathological changes of renal tissue, PAS staining was positive substance; kidney tissue of rats in model group showed extensive PAS staining positive material, mainly located in the central part of the lobule, glomerular basement membrane thickening, mesangial matrix increased, glomerular diffuse lesions, hyaline arteriolosclerosis, glomerular internal circular, elliptical or conical occasionally tuberous, individual glomerular obsolescence; and each treatment group and traditional Chinese medicine prevention of kidney of rats in the PAS group was weakly positive, western medicine group showed diffuse glomerular sclerosis, mesangial matrix and extracellular matrix increased, tubular atrophy; traditional Chinese medicine prevention group mesangial matrix increased slightly, glomerular basement membrane and renal tubular no lesion; high dose group showed mesangial matrix and mesangial cells increased, no change in the glomerular basement membrane, tubular no lesion; low dose traditional Chinese medicine group rats showed segmental glomerular sclerosis, mesangial matrix increased, basement membrane thickening.8. The changes of blood lipid in serum of rats in each group:(1)triglycerideExperimental results show that, compared with the normal group, model group rats serum triglyceride was significantly increased, there was significant difference between two groups (P<0.01); compared with the model group, the serum drug treatment group rats triglyceride decreased significantly (P<0.01), compared with the normal group also there was a significant difference (P<0.01), but the use of Chinese medicine treatment was no significant difference between the groups (P>0.05), indicating that the drug intervention can effectively reduce the content of triglyceride in serum of DN rats, can be close to normal level.(2) cholesterolExperimental results show that, compared with the normal group, serum cholesterol of rats in model group were significantly elevated, there was significant difference between two groups (P<0.01); compared with the model group, serum cholesterol in the drug treatment group decreased significantly (P<0.01), there was significant difference compared with the normal group (P<0.01), but the use of Chinese medicine was not significantly different between treatment groups (P>0.05), indicating that the drug intervention can effectively reduce serum cholesterol in DN rats, can be close to normal level.(3) and high density lipoproteinExperimental results show that, the content of high density lipoprotein in serum of rats in each group had no obvious change, there was no significant difference among the three groups (P>0.05).(4) and low density lipoproteinExperimental results show that, compared with the normal group, the serum low density lipoprotein in rats of model group increased obviously, there was significant difference between two groups (P<0.01);compared with the model group, the treatment group rats serum low density lipoprotein decreased significantly (P<0.01), and compared with normal group had significant difference (P<0.01), indicating that the drug intervention can effectively reduce the serum DN of rats with low density lipoprotein, but did not reach the normal level.9. Results the content of serum of rats in SREBP-lc, SREBP-2c(1) compared with the normal group, the content of SREBP-lc in serum of model group rats in the elevated significantly, there was significant difference between two groups (P<0.01); compared with the model group, the content of SREBP-lc in serum of drug use of rats in each group decreased significantly (P<0.01), but with normal group there is no significant difference (P>0.05), indicating that the drug intervention can effectively reduce the contents of SREBP-lc in serum of DN rats, but has not yet reached the normal level. Experimental results show that the effect of traditional Chinese medicine, preventive treatment group decreased the content of SREBP-lc in serum is the most obvious.(2) compared with the normal group, the content of SREBP-2c in serum of model group rats in the elevated significantly, there was significant difference between two groups (P<0.01); compared with the model group, the content of SREBP-2c in serum of rats in each group of drug use were decreased (P<0.01), in the use of drugs to the treatment group the content of serum SREBP-2c, traditional Chinese medicine drug use rats in the prevention and treatment group was most obvious decreased (P<0.01), indicating that the drug intervention can effectively reduce the contents of SREBP-2c in serum of DN rats, but has not yet reached the normal level. Experimental results show that the content of serum SREBP-2c, traditional Chinese medicine prevention treatment group rats in the most obvious.10. Results expression of rats in the renal tissue of SREBP-lc, SREBP-2c mRNA(1) compared with the normal group, the content of mRNA in SREBP-lc model group rats kidney tissue increased obviously, there was significant difference between two groups (P<0.01); compared with the model group, the content of mRNA SREBP-1c the use of drugs in the renal tissue of rats in each group were significantly lower (P<0.01), but with normal group had significant differences (P<0.01), and the traditional Chinese medicine prevention group, medication group had significant difference (P<0.05). Prompt medical intervention can effectively reduce the content of mRNA SREBP-lc DN in rat renal tissue, but has not yet reached the normal level.(2) compared with the normal group, the content of mRNA in SREBP-2c model group rats kidney tissue increased obviously, there was significant difference between two groups (P<0.01); compared with the model group, the content of mRNA SREBP-2c the use of drugs in the renal tissue of rats in each group were significantly lower (P<0.01), but with normal group had significant difference (P<0.01), and the traditional Chinese medicine prevention group, medication group had significant difference (P<0.05), indicating that the drug intervention can effectively reduce the content of mRNA SREBP-2c DN in rat renal tissue, but has not yet reached the normal level.11. Results the expression of SREBP-lc, SREBP-2c protein in renal tissue of rats in each group.(1) compared with the normal group, SREBP-lc model group rat kidney tissue protein expression was significantly increased, there was significant difference between two groups (P<0.01); compared with the model group, the expression of SREBP-lc rats using drugs in renal tissue protein decreased significantly (P<0.01), SREBP-lc prevention group rats treated with each drug group of traditional Chinese medicine in renal tissue protein expression in the lowest (P<0.01), indicating that the drug intervention can effectively reduce the SREBP-lc DN in rat renal tissue expression. (2) compared with the normal group, SREBP-2c model group rat kidney tissue protein expression was significantly increased, there was significant difference between two groups (P<0.01); compared with the model group, the expression of SREBP-2c in renal tissue of each drug group in the protein decreased significantly (P<0.01), but with normal group there was significant difference (P<0.01), SREBP-2c in renal tissue of each drug group Chinese medicine prevention group rats expressed proteins in the lowest (P<0.01), indicating that the (drug intervention can effectively reduce the SREBP-2c DN in rat renal tissue expression, expression but have not yet reached the normal group; high dose group and low dosage Chinese medicine group was higher than that of Western medicine group and traditional Chinese medicine prevention group.Conclusion:1. Using STZ once intraperitoneal injection molding induced early DN rats, can be replicated successfully establishing early DN model.2. Effect of Yiqi Jiedu active compound traditional Chinese medicine on early DN rats index improved, can effectively control the early DN rats blood glucose, glycosylated, urea nitrogen, serum lipid, renal function, urinary protein, serum β2change, protect the injured renal tissue, has obvious preventing and treating effect on early DN3. Possible mechanism of Yiqi Jiedu active preventive and therapeutic effects of Chinese medicine on DN and lipid toxin and steroid binding protein is closely related to regulatory element.

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