| Objective: lgA also known as Berger, is a kind of primaryglomerulopathy. This experiment is designed to study the therapeutic actionand possible mechanism of Qi supplementing and Yin nourishing drugs to IgAnephropathy and to provide experiment and theoretical basis of clinicaltreatment andexperimental studies by building model of nephropathy rat andtreat it with the drugs and benazepril, then detecting the occult blood of therat,24-hour urinary protein quantity, the red blood cell count,immunohistochemistry in order to examin the expression of α-SMA andTNF-α in rat kidney tissue.Method: In this experiment, using40SD rats with150g. All the ratswere fed adaptively for1week with standard diet, and then the ones whosenumber of RBC is negative would be chosen and randomly assigned to normalgroup, model group,benazepril group and Qi supplementing and Yinnourishing drugs group,10gerbils each. Establish the model of model group,benazepril group(B group) and Qi supplementing and Yin nourishing group(Qigroup). Inject400mg/kg of BSA every other day for eight weeks,100g/Laltogether. The rats were given subcutaneous injection of0.3ml of castor oiland0.1ml of CCl4for nine weeks;and in the6th weekend inject0.25g/L ofLPS by cauda vein,0.05mg for each. The normal group were given a gavageof distilled water of the same volume of4ml/kg, and were given subcutaneousinjection0.25g/L of LPS, once a week for nine weeks; and were injectedSodium Chloride Injection by cauda vein at the6th weekend.The rats were feed with common fodder for one weekand and of which24h of urine were collected with metabolic cages. The rats whose urine proteinobviously increased enter the stage of treatment. The Qi group were injected with Qi supplementing and Yin nourishing drugs everyday from the11thweek. The normal group and model group were given purified water of thesame volume for eight weeks. Collected24hours urine of each groups of ratswith metabolic cages before model, and at the4th weekend, the7th,the10th,the14th, and the18th. Then determine daily urinary protein. At the18thweekend, anesthetize rats with chloral hydrate. Rats were decapitated and theblood were collected and separated from the serum. Detection BUN and Scr inserum, collected the kidney tissue. Make HE，PAS，Masson，Jones silver stainand immunofluorescence test to detect the varition of pathology changes of theglomerulus. Determine the expression of α-SMA and TNF-α in the kidneytissue of model rats.Result:1The Comparison between urine protein quantitation of different groups ofrats in the same stage:Before model: no big difference between all groups.The4th weekend: comparing to the normal group, the other three groupsshow an distant increase in urine protein(P<0.05). There is no significantdifference (P>0.05).The7th weekend: comparing to the normal group, the other three groupsshow an distant increase in urine protein(P<0.05). There is no significantdifference (P>0.05).The10th weekend: comparing to the normal group, the other threegroups show an distant increase in urine protein(P<0.05). There is nosignificant difference between the three groups(P>0.05).The14th weekend: comparing to the normal group, the other threegroups show an distant increase in urine protein(P<0.05); Comparing to themodel group, the benazepril group，and the group of Qi supplementing andYin nourishing group show an distant decrease in urine protein(P<0.05); Thereis no significant difference between the benazepril group，and the group of Qisupplementing and Yin nourishing group (P>0.05).The18th weekend: comparing to the normal group, the other three groups show an distant increase in urine protein(P<0.05); Comparing to themodel group, the benazepril group，and the group of Qi supplementing andYin nourishing group show an distant decrease in urine protein(P<0.05); Thereis no significant difference between B group and Qi group(P>0.05).2The Comparison between urine red blood cell count of different groups ofrats in the same stage:The4th weekend: comparing to the normal group, the other three groupsshow an distant increase in urine red blood cell count(P<0.05). There is nosignificant difference (P>0.05).The7th weekend: comparing to the normal group, the other three groupsshow an distant increase in urine red blood cell count (P<0.05). There is nosignificant difference (P>0.05).The10th weekend: comparing to the normal group, the other threegroups show an distant increase in urine red blood cell count (P<0.05). Thereis no significant difference between the three groups(P>0.05).The14th weekend: comparing to the normal group, the other threegroups show an distant increase in urine protein(P<0.05); Comparing to themodel group, the benazepril group，and the group of Qi supplementing andYin nourishing show an distant decrease in urine red blood cell count(P<0.05);There is no significant difference between the benazepril group，andthe group of Qi supplementing and Yin nourishing (P>0.05).The18th weekend: comparing to the normal group, the other threegroups show an distant increase in urine red blood cell count(P<0.05);Comparing to the model group, B and Qi group show an distant decrease inurine red blood cell count (P<0.05); There is no significant difference betweenB group and Qi group(P>0.05).3Comparision between Scr and BUM of lgA ratsScr，BUN of the four groups do not vary significantly(P>0.05).4Observation of the pathological changes in the kidney4.1Light microscope:Normal group: structure of Glomerulus was integrated. No abnormal and significant change in mesentery and matrix.Model group: structure of Glomerulus was integrated. Mesangial cellsproliferated, and messegium increased.The benazepril group，and The group of Qi supplementing and Yinnourishing group: structure of Glomerulus was integrated. Messegiumincreased. Lesion reduced.4.2Immunity fluorescence:Normal group: immune complexes did not deposited or littele inglomerular mesangium.Model group: immune complexes present the pattern of"massivedistribution" along glomerular mesangium.The benazepril group,and The group of Qi supplementing and Yinnourishing group: different degree of immune complexes presented the patternof"massive distribution" along glomerular mesangium, lesser than modelgroup.5The expression of α-SMA，TNF-α in kidney tissue determined withimmunohistochemistry5.1expression of α-SMA protein in kidney tissueCompared with normal group, The positive expression of α-SMA proteinin kidney tissue of the other three groups enhanced(P<0.05); compared withmodel group, that of B group and Qi group weakened(P<0.05).5.2expression of TNF-α protein in kidney tissueCompared with normal group, the positive expression of TNF-α proteinin kidney tissue of the other three groups enhanced(P<0.05); compared withmodel group, that of B group and Qi group weakened(P<0.05).Conclusion:1Qi supplementing and Yin nourishing drugs could decrease urinary proteinin lgA nephropathy rat, thus protecting kidney function;2Qi supplementing and Yin nourishing drugs could decrease the synthesis ofα-SMA，TNF-α, preventing glomerular sclerosis and renal interstitial fibrosis,and reducing inflammation, thus postponing the progress of kidney disease. |
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