| Objective: To observe the clinical effect of Qingshen Granule on chronic renal failure patients with damp heat syndrome and the effect of serum leptin, Janus-activated kinase transducer and activator of transcription signal transduction pathway on the expression of JAK2 protein and STAT3 protein.Methods: According to the random number table, will meet the inclusion criteria of 68 cases of damp heat syndrome of CRF patients were divided into treatment group and control group with 34 cases in each, and 20 cases of normal subjects(for detection of serum leptin, JAK2 protein and STAT3 protein). In the course of the study, 8 patients were excluded, 5 cases in the treatment group, 3 cases in the control group, 60 cases were actually completed,including 29 cases in the treatment group and 31 cases in the control group. The treatment group and the control group were given retention enema with traditional Chinese medicine and the basic treatment of Western medicine, The treatment group added Qingshen granule with warm water, 3 times a day, 10 g each time for 12 weeks.To observe changes of clinical disease efficacy, TCM syndrome efficacy, TCM syndrome integral value,serum creatinine(Scr), glomerular filtration rate(GFR),serum leptin, JAK2 protein and STAT3 protein in both two groups before and after treatment.Results:1. The total effective rate of clinical disease therapeutic effect was 86.21% in the treatment group, it was 58.06% in the control group, the difference was significant between the two groups(P<0.05).2. The total effective rate of TCM syndromes was 82.76% in the treatment group, it was 54.84% in the control group, the difference was significant between the two groups(P<0.05).3. The two groups have no significant difference of TCM syndrome score before treatment(P>0.05). After treatment, the treatment group of TCM syndrome scoresignificantly reduced than before treatment(P<0.01), the control group of TCM syndrome score had statistical difference than before treatment(P<0.05); After treatment, the treatment group of TCM syndrome score compared with the same period control group was statistically significant difference(P<0.05).4. The two groups had no significant difference of Scr and eGFR before treatment(P>0.05); After treatment, the level of Scr in the treatment group were significantly lower than before treatment(P<0.01), the level of eGFR in the treatment group were significantly higher than before treatment; while the control group was not significantly decreased(P > 0.05); After treatment, the level of Scr in treatment group was lower than that in the same period control group, the level of eGFR in treatment group was higher than that in the same period control group,and the difference was statistically significant(P < 0.05).5. Before treatment,compared with the normal group, the levels of serum leptin, JAK2 and STAT3 in the treatment group and the control group were significantly increased(P<0.01);The two groups had no significant difference of serum leptin, JAK2 and STAT3 before treatment(P>0.05); After treatment, the levels of serum leptin,JAK2 and STAT3 in the treatment group and the control group were significantly lower than those before treatment(P<0.01); After treatment, the patients in the treatment group serum leptin, JAK2, STAT3 level compared with the control group decreased significantly(P<0.05, P<0.01).Conclusion:1. The levels of serum leptin were significantly increased in CRF patients with damp heat syndrome, the expression of JAK2 protein and STAT3 protein was increased.2. Qingshen granule could improve the clinical symptoms of CRF patients with damp heat syndrome, and improve the renal function.3. Qingshen granule could reduce serum leptin, JAK2 protein and STAT3 protein levels of CRF patients with damp heat syndrome, indicating that Qingshen granule may inhibit leptin mediated activation of JAK / STAT signal transductionpathway,plays an important role in improving renal function and slows the progression of renal fibrosis.4. Patients taking Qingshen granule had no adverse reaction, safety indexs was measured without obviously abnormal, indicating that the drug had a good safety.Objective: By establishing the rat model of unilateral ureteral obstruction(UUO), to observe the changes of JAK/STAT signaling pathway by leptin in UUO model rats, and to investigate the mechanism of renal fibrosis; To observe the intervention effect of Qingshen Granule on the JAK/STAT signaling pathway by leptin in UUO model rats and clarify the mechanism of anti renal fibrosis.Methods: 72 male SD rats were selected and the adaptive feeding for one weeks. After one weeks, them were randomly divided into 6 groups by random sampling, including normal group, model group, Qingshen Granule high-dose group, Qingshen Granule medium-dose group, Qingshen Granule low-dose group and valsartan group, with 12 rats in each group. In addition to 12 rats in the normal group were separated out the left ureter without ligation,the other 60 rats were established as UUO model by theoperative method. Animals in each group were given medicine on the first day after modeling. The low, medium and high dose groups of Qingshen Granule: the Qingshen granule aqueous solution were respectively treated with 4.0g/kg?d 、 8.0g/kg?d and16.0g/kg?d;Valsartan group: the suspension of valsartan capsule was administered by20mg/kg·d; Normal group and model group were given warm water to fill the stomach once a day, each time 2ml, for 12 weeks. To detect 24 hour urinary protein(24h Upro)and Scr, BUN,and to detect the serum leptin level by ELISA method; To detect the content of leptin receptor(OB-R), phosphorylated JAK2 protein(p-JAK2), phosphorylated STAT3 protein(p-STAT3), nuclear factors-κBp65(NF-κBp65) and Monocytechemotatic protein-1(MCP-1) in kidney tissue by western blot method; To detect the expression levels of leptin, JAK2 protein, STAT3 protein, E- calcium mucin(E-cadherin) and alpha smooth muscle actin(α--SMA) m RNA in renal tissue by RT-PCR method; To detect the expression of Col- IV and FN in renal tissue by Immunohistochemical method; To observe renal pathological changes by HE and PAS staining.Results:1. Comparison of 24 h Upro,BUN and Scr: Compared with the normal group,the levels of 24 h Upro,BUN and Scr in the model group and drug treatment group rats were significantly higher(P<0.05, P<0.01); Compared with the model group, the levels of 24 h Upro,BUN and Scr in the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.05, P<0.01); The middle dose group decreased most significantly in each treatment groups of Qingshen granule; Compared the levels of 24 h Upro,BUN in valsartan group with Qingshen granule high and middle dose group rats, there was no significant difference between the two groups(P>0.05),compared the levels of Scr in valsartan group with each treatment groups of Qingshen granule, there was no significant difference(P>0.05);2. Comparison of serum leptin of rats: Compared with the normal group, the level of serum leptin in the model group and drug treatment group rats were significantly higher(P<0.01); Compared with the model group, the levels of serum leptin in the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.01), the middle dose group decreased most significantly; Compared the level of serum leptin in valsartan group with Qingshen granule middle dose group rats, the difference was not statistically significant(P>0.05).3. Comparison of OB-R、p-JAK2、p-STAT3、NF-k Bp65、MCP-1 protein in renal tissue of rats in each group: Compared with the normal group, the levels of OB-R、 p-JAK2、p-STAT3、NF-k Bp65、MCP-1 in the model group and drug treatment group rats were significantly higher(P<0.05,P<0.01); Compared with the model group, Compared with the model group, the levels of OB-R、p-JAK2、p-STAT3、NF-k Bp65、MCP-1 in the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.01), the middle dose group decreased most significantly; Compared the levels of OB-R、MCP-1 in valsartan group with Qingshen granule high dose group rats, there were no significant difference(P>0.05),compared the levels of p-JAK2、p-STAT3、NF-k Bp65 in valsartan group with Qingshen granule middle dose group rats, the difference was not statistically significant(P>0.05).4. Comparison of leptin、JAK2、STAT3、E-cadherin、α-SMA mRNA in renal tissue of rats in each group: Compared with the normal group, the levels of leptin、JAK2、STAT3 、 α-SMA m RNA in the model group and drug treatment group rats were significantly higher(P<0.01),the level of E-cadherin in the model group and drug treatment group rats were significantly lower(P<0.05,P<0.01); Compared with the model group, the levels of leptin、JAK2、STAT3、α-SMA m RNA in the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.01), the middle dose group decreased most significantly; Compared with the model group, the levels of E-cadherin in the each dose of Qingshen granule group and valsartan group were significantly higher(P<0.05,P<0.01), the middle dose group increased most significantly; Compared the levels of leptin、JAK2、E-cadherin、α-SMA in valsartan group with Qingshen granule high and middle dose group rats, there were no significant difference(P>0.05),compared the levels of STAT3 in valsartan group with Qingshen granule middle dose group rats, the difference was not statistically significant(P>0.05).5. Comparison of immunohistochemical results of rats in each group: The normal group of renal tubular epithelial cells and interstitial cells expressed a small amount of FN and Col-Ⅳ.Compared with the normal group, the levels of FN and Col-Ⅳ in the model group and drug treatment group rats were significantly higher(P<0.05,P<0.01); Compared with the model group, the levels of FN and Col-Ⅳin the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.05,P<0.01), the middle dose group decreased most significantly; Compared the levels of FN and Col-Ⅳ in valsartan group with Qingshen granule high and low dose group rats, there were no significant difference(P>0.05),6. Comparison of renal tubular interstitial injury of rats in each group: Scores of the model group and drug treatment group were significantly higher than the normal group(P<0.01); Compared with the model group, the scores in the each dose of Qingshen granule group and valsartan group were significantly lower(P<0.05,P<0.01), the middle dose group decreased most significantly; Compared the scores in valsartan group with Qingshen granule middle dose group, the difference was statistically significant(P<0.05), compared with the high and low dose group, there were no significant difference(P>0.05).Conclusion:1.The activation of JAK/STAT signaling pathway mediated by leptin was involved in the process of renal interstitial fibrosis in UUO rats.2. Qingshen granule could reduce the levels of 24 h Upro,BUN and Scr in UUO rats.3. Qingshen granule could reduce renal damage of the renal interstitial fibrosis in rats, has the role of anti renal fibrosis.4. Qingshen granule could inhibit the activity of JAK/STAT signaling pathway mediated by leptin, thereby reducing NF-κB activity and induced inflammatory reaction, thereby inhibiting renal tubular EMT, reducing the degree of renal fibrosis. |
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