The Intervention Of Huayu Tongluo Traditional Chinese Medicine To Diabetic Nephropathy Rats And Its Effect On Kidney Desmin Expression

Objective: Diabetic nephropathy (DN) refers to as diabetes can causeglomerular lesions, is a serious complication of diabetes develops to a certainstage. The study by intraperitoneal injection of streptozotocin (STZ) toestablish the model of DN rats, Huayu Tongluo intervention, by observing themorphological changes of renal tissue in rats, and in general, quantitativemeasurement of24hours urinary protein, blood regular test, bloodbiochemical index, the expression level of desmin, study of Huayu Tongluomedicine role, and to explore its possible mechanism.Methods:80male SD rats were randomly selected15for the normalcontrol group (C), the remaining rats were fasted for12hours or more, asingle intraperitoneal injection of STZ (55mg/kg), measured rats’ tail veinrandom blood glucose≥16.7mmol/L after72hours for successful modeling.The model rats were randomly divided into model group (M), the HuayuTongluo group (Q), the Huayu group (H), Tongluo group (T), the irbesartangroup (E), the corresponding drug orally to16weeks. Rats’24-hour urinewere collected at the end of16weeks. Fasted for more than12hours, the rats’blood were drew under anesthesia condition for testing serum biochemicalmarkers, and specimens from renal tissue for light microscopy, electronmicroscopy, immunohistochemistry and PCR detection.Results:1Rats general statusNormal control rats were generally in good condition; rats after STZinjection molded, typically drank more, ate more, urinated more, becamethinner and the most significant in the model group. During treatment, HuayuTongluo group one died, irbesartan group two died, model group six died. 224-hour urinary total protein (24h UTP)The results showed: Total urinary protein level of24hours in eachtreatment group with the normal control group and with model group weredifferent (P<0.05), significantly higher than the normal control group, butsignificantly lower than the model group; compared among treatment groups,Huayu group24-hour urine total protein was significantly higher than theHuayu Tongluo group, the Tongluo group and irbesartan group (P <0.05).3Blood test resultsIn blood regular test, compared with the normal control group, the HuayuTongluo group, the Huayu group, the Tongluo group and the irbesartan group,the platelet count in the model group increased significantly (P<0.05);compared with the model group and the irbesartan group, the large plateletratio (P-LCR) in the normal control group, the Huayu Tongluo group, HuayuGroup, Tongluo group was significantly lower (P <0.05).In blood biochemical test, compared with the normal control group, TGlevels in the modeling rats in each group were significantly increased (P<0.05),the Huayu Tongluo group, the Tongluo group and the irbesartan groupcompared with the model group and the Huayu group were significantlylower(P<0.05); compared with the normal control group; TP level in themodel group, the Huayu Tongluo group, the Huayu group, the Tongluo groupwas significantly lower (P<0.05), TP level in the model group, the HuayuTongluo group, the Huayu group and the Tongluo group was not statisticallysignificant (P>0.05); TP level between the irbesartan group and the normalcontrol group was not statistically significant (P>0.05); the modeling groupsALB level was significantly reduced compared with the control group(P<0.05), ALB level in each treatment group compared with the model groupwas significantly higher (P<0.05), among the treatment groups was notstatistically significant (P>0.05).4Diabetic Cataract (DC)The results showed: Normal control group found no cataract rats; modelgroup all cataract rats; the incidence of cataract in the Huayu Tongluo group is 45%; the incidence of cataract in the Huayu group is circulation group is42%;the incidence of cataract in the Tongluo group is circulation group is92%; the incidence of cataract in the irbesartan group is90%. Apparently, theHuayu Tongluo group and the Huayu group had less diabetic cataract rats thanthe Tongluo and irbesartan group.5Renal tissue morphology observations5.1Light microscopy observationsNormal control group rats had clear and normal kidney tissue structure;model group rats glomerular hypertrophy, renal capsule was slit-like evenballoon adhesions, obvious thickening of the basement membrane, mesangialhyperplasia; compared with the model group, each treatment group wassignificantly reduced pathological changes; among the treatment groups, theHuayu group pathology performance is slightly heavier than the othertreatment groups.5.2Electron microscopy observationsNormal control group rats had clear and normal kidney tissue structure;model group rats heterogeneity of glomerular basement membrane thickening,increased mesangial matrix, podocyte foot processes were widely fusion;compared with the model group, each treatment group pathology changedsignificantly reduced; among the treatment groups, the Huayu grouppathology performance is slightly heavier than the other treatment groups.6Kidney tissue Desmin immunohistochemistry assayThe results showed: Normal control group showed a small amount ofexpression of desmin in glomerular mesangial cells, podocytes showed noexpression; compared with normal control group, modeling group desminexpression was significantly increased (P<0.05) in both podocytes andmesangial cells; compared with the model group, the treatment group desminexpression significantly decreased (P <0.05); among the treatment groups, theHuayu group and the Huayu Tongluo group, the Tongluo group, the irbesartangroup were significantly different (P <0.05).7Real-TimePCR detection of renal tissue Desmin mRNA expression The results showed: Compared with the normal control group, modelinggroups desmin mRNA expression was significantly increased (P <0.05) in therenal tissue; compared with the model group, desmin mRNA expression ineach treatment group was significantly reduced (P<0.05); among the treatmentgroups, the Huayu group and the Huayu Tongluo group, the Tongluo group,the irbesartan group were significantly different (P <0.05).Conclusions:1Huayu Tongluo medicine can improve the general status of diabeticnephropathy rats, decrease the urine protein, maintain the plasma proteincontent and relieve pathology of diabetic nephropathy, these aspects of therole similar to irbesartan.2Huayu Tongluo medicine can decrease the number of platelet, reduceits activity, thereby preventing thrombosis, the effect is superior to that ofirbesartan.3Huayu Tongluo medicine can decrease plasma triglyceride levels,improve lipid metabolism disorder in diabetes, role in Tongluo medicine issuperior to Huayu medicine.4Huayu Tongluo medicine can reduce the incidence of diabetic cataract,and Huayu medicine is superior to Tongluo medicine and irbesartan.5Huayu Tongluo medicine can reduce desmin expression in the kidneyof diabetic nephropathy rats, suggesting its therapeutic effect on DN may berelated to protecting podocytes.6In conclusion，Huayu Tongluo medicine is superior to Huayu medicineand Tongluo medicine separately.