| Objective:To observe the effect of Yishenkang granule on UTRF content and kidneyultrastructure of DM rat models, and to investigate the therapeutic mechanismof Yishenkang granule on early DN.Methods:120male SPF SD rats,11rats of which were randomly selected as normal groupwithout any processing.After the rest of the rats were provided with no foodbut water for12hours, DM rats were induced by a single intraperitonealinjection of STZ at the concentration of53mg/kg.72hours later, these injectedrats were drew blood through eye sockets to measure blood glucose levels. Thenthe blood glucose values of which were equal or greater than16.7mmol/L wererandomly divided into model group (n=16), positive drug control group (n=16),Yishenkang granule low dose group (n=16), Yishenkang granule middle dose group(n=17) and Yishenkang granule high dose group (n=18).The11rats of normal group without STZ injection were also measured theblood glucose levels through eye sockets blooding. Rats of normal group and modelgroup were intragastric administration with normovolemic physiological salinesolution. Positive drug control group was intragastric administration withRamipril suspension. Yishenkang granule low dose group, middle dose group and highdose group were respectively intragastric administration with three differentconcentrations of Yishenkang granule. Drug cycles of each group were8weeks.Intragastric administration at the end of4,6and8weeks,24hours urineof each rat was respectively collected with metabolism cage,2ml of which wastaken out for-20℃refrigerator storage. The content of UTRF was detected byELISA method. After collecting urine at the end of8weeks, each rat was hocusedwith intraperitoneal injection of25%urethane. Blood sample of each rat was obtained from abdominal aorta. Two kidneys of each rat were removed and weighedunder intraperitoneal anesthesia, getting rid of the capsule and the surroundingadipose tissue. Then the kidney index was calculated.1mm3renal cortex tissueof each left kidney was removed and immersed in2.5%glutaric dialdehyde formaking observation specimen of electron microscope.Results and Analysis:1. At the end of4weeks, UTRF content of model group was higher than thatof normal group (p＜0.01), which illustrated that DM rats could turn up clinicalmanifestation of early DN at four weeks’ course of the disease. After fourweeks’ treatment, UTRF content of positive drug control group was lower thanthat of model group, there were statistical differences between them (p＜0.05),which accounted for that four weeks’treatment of Ramipril could reduce thecontent of UTRF. Although UTRF content of Yishenkang granule three differentdoses groups presented a declined tendency, there were no obviously statisticaldifferences compared to model group (p＞0.05). The results may be due to thatTraditional Chinese Medicine worked slowly and medication cycles were too short.2. At the end of6and8weeks, UTRF content of model group was higher thanthat of normal group, there were significant differences between them (p＜0.01),and model group’s UTRF content of8weeks was higher than that of6weeks, whichexplained that kidney injury was aggravating gradually with the course of thedisease extended. UTRF content of positive drug control group, Yishenkanggranule low dose, middle dose and high dose group were significantly lower thanthat of model group, there were significant differences compared to model group(p＜0.01). The results pointed out that Ramipril and Yishenkang granule couldreduce UTRF content. UTRF content of Yishenkang granule high dose group was lowerthan that of positive drug control group, there were statistical differencesbetween them (p＜0.05), Although UTRF content of Yishenkang granule low andmiddle dose groups were lower than that of positive drug control group, therewere no statistical differences between them (p＞0.05). The results illustrated that the function of reducing UTRF content of Yishenkang granule high dose groupwas better than Ramipril. UTRF content of Yishenkang granule high dose groupwas lower than that of normal group, but there were no statistical differencesbetween them (p＞0.05), which explained that Yishenkang granule high dose groupcould reduce UTRF content, but still could not return to normal level.3. For Yishenkang granule three different doses groups, UTRF content of6and8weeks was lower than that of4weeks, there were obviously statisticaldifferences between them (p＜0.01), UTRF content of8weeks was lower than thatof6weeks, there were statistical differences between them (p＜0.05), whichexplained that with medication cycle and concentration extended, Yishenkanggranule could reduce UTRF content significantly.And the function of Yishenkanggranule high dose group at the end of8weeks was the most significant.4. At the end of8weeks, the right kidney index of model group was higherthan that of normal group, there was significant difference compared to normalgroup (p＜0.01). The results illustrated that early DN could turn up kidneyhypertrophia. The right kidney index of Yishenkang granule high dose group waslower than that of model group, there were statistical differences between them(p＜0.05), the right kidney index of Yishenkang granule low dose group and middledose group were lower than that of model group, but there were no statisticaldifferences (p＞0.05). The results accounted for that all three different dosesof Yishenkang granule could improve renal hypertrophy, and the effect ofYishenkang granule high dose group was the most significant. The right kidneyindex of Yishenkang granule middle dose group and high dose group were lowerthan that of positive drug control group, but there were no statisticaldifferences (p＞0.05). The results explained that the function of improvingrenal hypertrophy of Yishenkang granule middle dose group and high dose groupwere equal to Ramipril.The right kidney index of positive drug control group,Yishenkang granule low dose,middle dose and high dose groups were higher thanthat of normal group, there were significant differences between them (p＜0.01).The results explained that after eight weeks’ treatment, renal hypertrophy of both Ramipril and Yishenkang granule group improved, but these kidneys stillcould not return to normal sizes.5. We drew a conclusion from statistic analysis of the experimental datathat there was linearly positive correlation between both UTRF and UmAlbmeasurement indicators of model group at the end of6and8weeks. The correlationcoefficient of6weeks was0.859, the correlation coefficient of8weeks was0.760, which illustrated that the increasing of UTRF and UmAlb presentedlinearly positive correlation.6. Through scanning electron microscope we can see that glomerulus of modelgroup presented dissolved state, protuberances of podoryte fused together,glomerular capillary vessel endothelial fenestra and slit diaphragmdisappeared, endothelial cells atrophied. Each treatment group presentedprotuberance of sertoli cells to varying degrees, which were similar comparedto normal group. Part of these protuberances fused together, endothelialfenestra and slit diaphragm of sertoli cells disappeared partly. The resultsillustrated that kidney ultrastructure injury of Yishenkang granule threedifferent doses groups were slighter than that of model group.Conclusions:1. Early DN models were induced successfully.2. Both increasing of UTRF and UmAlb of early DN presented linearly positivecorrelation.3. Yishenkang granule has the function of inhibiting kidney hypertrophiaof early DN rats.4. Yishenkang granule could prevent and cure early DN through reducing thecontent of UTRF, and the therapeutic effect of high dose group was equal toRamipril.5. Yishenkang granule has the function of reducing early DN rats’kidneyultrastructure damages.In conclusion, Yishenkang granule has the function of preventing and curing early DN. |
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