| Objective: To observe the change of serum interleukin-17(IL-17),nuclear factors-κBp65(NF-κBp65) in peripheral blood mononuclear cells(PBMC),serum collagen typeⅢ (Col-Ⅲ) of chronic renal failure patients with damp-heat syndrome,and theintervention effect of Qingshen granule.Methods: sixty-eight patients with chronic renal failure of damp-heat syndromewere randomly assigned to treatment group and control group using random digittable.Choose thirty cases of healthy physical examination as normal group.Processof seven patients were lost to follow-up,four cases in treatment group and three casesin control group,the actual completed in sixty-one cases, of which the treatment groupthirty cases, the control group thirty-one cases.The treatment group including eighteenmale and twelve female,average age (53.20±9.41)years.The control group includingnineteen male and twelve female,average age (49.06±12.53)years.Treatment groupand control group were given basic treatment of western medicine and TraditionalChinese Medicine(TCM) retain enema.The same time,treatment group was added withQingshen granule, three times a day,one bag each time,the course of eightweeks.Detection of before and after treatment in treatment group and control groupserum reatinine(Scr),IL-17,Col-Ⅲ levels,NF-κ Bp65in PBMC and estimatedglomerular filtration rate(eGFR)，and observe the changes of TCM symptom integralin both group before and after treatment.Detection the levels of normal group’s serumIL-17,Col-Ⅲ,NF-κBp65in PBMC.Results:1.Comparison of clinical curative effect: treatment group curative effect thetotal efficiency was86.67%,58.06%in the control group, there was significantdifference between two groups (P<0.05).2.Comparison of TCM Syndrome efficacy:treatment group TCM syndrome total efficiency was86.67%, and control group was 45.16%, two groups were significant difference (P<0.01).3.TCM syndrome scorecomparison:Before treatment,treatment group TCM syndrome score compared with thecontrol group, no significant difference (P>0.05).After treatment, TCM syndromescore was significantly lower than that before treatment (P<0.01).The control groupafter treatment of the TCM syndromes score compared with before treatment, nosignificant difference (P>0.05).After treatment,the treatment group TCM syndromescore was significantly lower than the same period in control group(P<0.01).4.Comparison of Scr and eGFR:(1) Before treatment, treatment group Scrlevel compared with the control group had no statistical significance (P>0.05).Aftertreatment, the level of Scr significantly decreased compared with those beforetreatment (P<0.01).The control group level of Scr group after treatment compared withbefore treatment, no significant difference (P>0.05). After treatment, treatment groupScr level lower than the same period in the control group(P<0.05).(2)Before treatment,treatment group eGFR level compared with the control group had no statisticalsignificance (P>0.05).After treatment, the level of eGFR was significantly higher thanthat before treatment (P<0.01).The control group level of eGFR after treatmentcompared with before treatment, no significant difference (P>0.05). eGFR levels aftertreatment in treatment group was higher than that of the control group(P<0.05).5.Comparison of serum IL-17levels:Before treatment, treatment group theserum level of IL-17compared with the normal group, the difference was significant(P<0.01). Before treatment,control group the serum level of IL-17was higher thannormal group, there was significant difference (P<0.01).But the treatment group beforetreatment serum IL-17level compared with the control group, no significantdifference (P>0.05).After treatment, the treatment group serum IL-17levels lower thanthose before therapy (P<0.05).The serum IL-17level of control group after treatmentcompared with before treatment, no significant difference (P>0.05).Serum IL-17levelsafter treatment in treatment group was lower than the control group in the same period(P<0.05).6.Comparison of NF-κ Bp65in PBMC:The treatment group before treatment PBMC NF-κ Bp65content was higher than that of the normal group, thedifference was significant (P<0.01).The control group before treatment PBMC NF-κBp65content was higher than that of the normal group, the difference was significant(P<0.01).But comparison of the treatment group and the control group content of NF-κ Bp65in PBMC group before treatment, no significant difference (P>0.05).Aftertreatment, the PBMC of NF-κ Bp65content was significantly lower than that beforetreatment (P<0.01).The control group after treatment PBMC NF-κ Bp65contentcompared with before treatment, no significant difference (P>0.05).Aftertreatment,treatment group the PBMC of NF-κ Bp65content was significantly lowerthan the same period in the control group (P<0.01).7.Comparison of the level of serumCol-Ⅲ:Before treatment, treatment group and the control group serum Col-III levelswere higher than that of the normal, there was a significant difference (P<0.01).Beforetreatment, treatment group compared with control group of serum Col-Ⅲ level, nosignificant difference (P>0.05).After treatment, treatment group the level of serumCol-Ⅲ significantly decreased compared with those before treatment (P<0.01).Thecontrol group after treatment the serum Col-III levels compared with before treatment,no significant difference (P>0.05).The serum Col-III levels after treatment intreatment group were lower than those of the same period in the controlgroup(P<0.01).Conclusion:1.Qingshen granule can improve the clinical symptoms in CRF patientswith damp heat syndrome, it can decrease the levels of Scr and elevate levels ofeGFR.2.Levels of serum IL-17, Col-Ⅲ and NF-κ Bp65in PBMC in CRF patientswith damp heat syndrome are increase.3.Qingshen granule can reduce the levels ofserum IL-17, Col-Ⅲ and NF-κ Bp65in PBMC,it is description of Qingshengranule with better prevention and treatment of renal fibrosis.4.After treatment safetyindex are detected, there is no abnormal change, adverse drug reaction does not appearin the course of treatment,it is description of Qingshen granule with good security. Objective: Through the detection of unilateral ureteral obstruction (UUO) inducedrenal interstitial fibrosis (RIF) in the peripheral blood of rats T helper lymphocytes(CD4+T cells) and suppressor T lymphocytes (CD8+T cells) ratio, ratio of Th17cellsand kidney tissue of interleukin-17(IL-17), nuclear factor κ B-p65(NF-κBp65),collagen type Ⅲ(Col-Ⅲ) expression level to observe of UUO rat the existence ofdisorder of immune function and inflammatory mediators release of excessive ornot,and to observe the intervention effect of Qingshen granule.Methods: Totally60healthy male SD rats were randomly divided into5groups byrandomized block method: Qingshen granule group, Bailing capsule group, modelgroup, sham operation group, normal group,12rats in each group. Before theexperiment the rats blood urea nitrogen (BUN), serum creatinine (Scr) levels weredetected, no statistically significant difference between the groups (P>0.05). Qingshengranule group, Bailing capsule group and model group were prepared by using UUO inrat model of renal interstitial fibrosis,sham operation group had the same operationpath separation ureter but not ligation. Qingshen granule group in Qingshen granule0.6g/100g dissolved in4ml water by gastric perfusion.Bailing capsule group with0.03g/100g Bailing capsule dissolved in4ml water by gavage.The model group, shamoperation group and normal group were intragastric administration with4ml warmwater. The rats intragastrically once daily, for3weeks. After intragastric administrationover,takken the experimental rats abdominal aortic blood and the left kidney,before theoperation all rats leaved24hour urine volume. Biochemical method for BUN, Scrdetection and24hour urinary protein quantitative. Used the flow cytometry technologyto detecte peripheral blood CD4+/CD8+, Th17cell ratio. HE and Masson staining forpathological examination. Immunohistochemical method was used to detect theexpression of IL-17, Col-Ⅲ and NF-κ Bp65in renal tissue. Results:1.Comparison of BUN and Scr level:(1)Comparison between pretherapyBUN rats in each group, no significant difference (P>0.05).After treatment, Qingshengranule group, Bailing capsule group and model group rats BUN level were increased,it compared with the normal group and sham operation group with significantdifference(P<0.01).Compared with the normal group, the level of BUN in rats of thesham operation group was no significant difference (P>0.05).Qingshen granule groupand Bailing capsule group level of BUN were lower than the model group, there wassignificant difference (P<0.01).Qingshen granule group rats BUN levels was lowerthan Bailing capsule group, the difference was statistically significant(P<0.05).(2)Comparison of the level of Scr in each group before treatment, nosignificant difference (P>0.05).After treatment, Qingshen granule group, Bailingcapsule group and model group rats Scr level were increased, with significantdifference from the normal group and sham operation group (P<0.01).Comparison ofScr levels in sham operation group rats and normal rats, there was no significantdifference (P>0.05).Qingshen granule group and Bailing capsule group level of Scrwere lower than the model group, there was significant difference (P<0.01).Qingshengranule group Scr levels was lower than Bailing capsule group, the difference wasstatistically significant (P<0.05).2.Comparison of24hour urinary proteinquantitative:Bailing capsule group and model group rats24hour urinary proteinquantitative were higher than that in normal group and sham operation group, therewas significant difference (P<0.01).Qingshen granule group rats24hour urinaryprotein quantitative was higher than normal group, there was significant difference(P<0.01).Qingshen granule group rats24hour urinary protein quantitative was higherthan the sham operation group, the difference was statistically significant(P<0.05).Sham operation group rats24hour urinary protein quantitative comparedwith the normal group, the difference was not statistically significant(P>0.05).Qingshen granule group and Bailing capsule group rats24hour urinaryprotein quantitative were lower than the model group, there was significant difference (P<0.01).Qingshen granule group rats24hour urinary protein quantitative was lowerthan Bailing capsule group, the difference was statistically significant (P<0.05).3.Comparison of peripheral blood CD4+/CD8+:Qingshen granule group rats peripheralblood CD4+/CD8+was higher than the normal group and the sham operation group, thedifference was statistically significant (P<0.05).Bailing capsule group and modelgroup of rats peripheral blood CD4+/CD8+were higher than that in normal group andsham operation group, there was significant difference (P<0.01).Compared the normalgroup of peripheral blood CD4+/CD8+in rats with the sham operation group, nosignificant difference (P>0.05).Qingshen granule group of peripheral bloodCD4+/CD8+was lower than the model group, there was significant difference(P<0.01).Bailing capsule group of peripheral blood CD4+/CD8+was lower than themodel group, the difference was statistically significant (P<0.05).Qingshen granulegroup rats peripheral blood CD4+/CD8+was lower than Bailing capsule group, therewas significant difference (P<0.01).4.Comparison the proportion of peripheral Th17cells:Qingshen granule group, Bailing capsule group and model group rats peripheralTh17cell ratio were higher than that in normal group and sham operation group, therewas significant difference (P<0.01).Sham operation group rats peripheral Th17cellratio compared with the normal group, the difference was not statistically significant(P>0.05).Qingshen granule group rats peripheral Th17cell ratio lower than the modelgroup, there was significant difference (P<0.01).Bailing capsule group rats peripheralTh17cell ratio lower than the model group, the difference was statistically significant(P<0.05).But Qingshen granule group rat peripheral Th17cell ratio was lower thanBailing capsule group, the difference was statistically significant (P<0.05).5.Comparison of immunohistochemical IL-17staining:Qingshen granule group,Bailing capsule group and model group rats IL-17semi quantitative score were higherthan that in normal group and sham operation group, there was significant difference(P<0.01).Sham operation group rats IL-17semi quantitative score compared with thenormal group, the difference was not statistically significant (P>0.05).Qingshen granule group and Bailing capsule group rats IL-17semi quantitative scores werelower than the model group, there was significant difference (P<0.01).Qingshengranule group rats IL-17semi quantitative score was lower than Bailing capsule group,the difference was statistically significant (P<0.05).6.Comparison ofimmunohistochemical NF-κ Bp65staining:Qingshen granule group, Bailing capsulegroup and model group rats NF-κ Bp65coloring semi quantitative score werehigher than that in normal group and sham operation group, there was significantdifference (P<0.01).Sham operation group rats NF-κ Bp65coloring semiquantitative score compared with the normal group, the difference was not statisticallysignificant (P>0.05).Qingshen granule group and Bailing capsule group rats NF-κBp65coloring semi quantitative scores were lower than the model group, there wassignificant difference (P<0.01).Qingshen granule group rats NF-κ Bp65coloringsemi quantitative scores were lower than Bailing capsule group, there was significantdifference (P<0.01).7.Comparison of immunohistochemical Col-Ⅲ staining:Bailingcapsule group and the model group rats Col-Ⅲ coloring semi quantitative score werehigher than that in normal group and sham operation group, there was significantdifference (P<0.01).Qingshen granule group rats Col-Ⅲ staining Semi quantitativescore was higher than normal group and sham operation group, the difference wasstatistically significant (P<0.05).Sham operation group rats Col-Ⅲ coloring semiquantitative score compared with the normal group, the difference was not statisticallysignificant (P>0.05).Qingshen granule group and Bailing capsule group rats Col-Ⅲcoloring semi quantitative scores were lower than the model group, there wassignificant difference (P<0.01).Qingshen granule group rats Col-Ⅲ staining Semiquantitative score was lower than Bailing capsule group, the difference wasstatistically significant (P<0.05).Conclusions:1.The inflammatory immune mediated mechanisms are involved in theoccurrence of renal interstitial fibrosis.2.Qingshen granule can reduce renal fibrosis in rats serum creatinine, urea nitrogen level, and it can reduce24h urine proteinquantity.3.Qingshen granule can reduce the renal pathological damage of renal fibrosisin rats, and it can reduce inflammation.4.Qingshen granule can regulate the disorder ofimmune function, and reduce inflammation medium IL-17release, inhibition of NF-κ B signal pathway, reduce NF-κ Bp65release, down regulated the expression ofrats with renal interstitial renal fibrosis Col-III, reduced renal interstitial extracellularmatrix (ECM) deposition, thus plays renal fibrosis antagonist (RF) function. |
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