艾灸延缓肾小管间质纤维化的作用与机制初探OA

Objective To investigate the mechanism of moxibustion intervention on renal tubulointerstitial fibrosis(TIF)in mice with unilateral ureteral obstruction(UUO).Methods A TIF mouse model was established using the UUO method in C57BL/6J mice.The mice were divided into Sham operation(Sham)group,Sham operation plus moxibustion(Sham+Moxa)group,Model group,and Model plus moxibustion(Moxa)group.Pathological changes in renal tissue were observed using Hematoxylin-Eosin,Sirius Red and Masson's staining.Collagen volume fraction(CVF)was measured,and immunohistochemistry was performed to detect α-smooth muscle actin(α-SMA),type Ⅰ collagen(Col Ⅰ),so as to evaluate the degree of TIF.The immunofluorescence method was used to detect hypoxia-inducible factor-1α(HIF-1α)in renal tubules to assess the ischemic-hypoxic state of renal tubules;Thrombospondin(TSP-1)and platelet endothelial cell adhesion molecule-1(PECAM-1/CD31)were detected to evaluate vascular injury;Double staining with the anti-aging protein Klotho/CD31 was detected to assess the Klotho expression in the endothelial cells of peritubular capillaries(PTC)around the renal tubules.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to measure the mRNA expression of fibrosis-related factors Snail family transcriptional repressor 1(Snail1)and CCAAT enhancer binding protein delta(Cebpd).Results Moxibustion could significantly improve renal tubular atrophy and luminal stenosis,reduce extracellular matrix hyperplasia and interstitial collagen fiber expression,and decrease CVF.Immunohistochemistry showed that moxibustion could reduce the positive expression of α-SMA and ColⅠ in the renal tubulointerstitium.Immunofluorescence revealed that moxibustion could decrease the positive expression of HIF-1α and TSP-1 in renal tubular epithelial cells and increase the positive expression of CD31 and Klotho in PTC endothelial cells.RT-qPCR demonstrated that moxibustion could downregulate the mRNA expression of Snail1 and Cebpd in renal tissue.Conclusions Moxibustion delays the progression of TIF by upregulating Klotho expression in PTC endothelial cells,alleviating ischemia and hypoxia in renal tubular epithelial cells,and inhibiting the expression of fibrosis-related factors.