| ObjectivesIn this study,using senescence-accelerate mouse prone 8（SAMP8）,an animal model of accelerated aging,we observed the pathological changes in aged mice kidneys,and the expression of the Sonic Hedgehog signaling pathway-related molecules was analyzed as well.Meanwhile,the effects of moxibustion at the meridian point BL-23（Shen-shu）on renal fibrosis in mice and Sonic Hedgehog proteins（Shh,Smo,and Gli1）proteins were investigated.It is hoped that our results will be able to provide new entry points for related research on the prevention and treatment of renal damage and renal function decline.MethodsIn our study,we used 10-month-old SAMP8（SPF-grade）as a model and senescence-accelerated mouse/resistance 1（SAMR1）of the same age as a control.For the moxibustion treatment group,moxibustion with moxa sticks is performed 2-3 cm above the bilateral BL-23（on the level of the lower border of the spinous process of the 2nd lumbar vertebra,and 3mm lateral to the posterior midline）,for 15 min each time,1 time a day,6 times constituted one course,four courses were required.The mice of the model group and the control group did not receive the experimental treatment but should be grasped and restrained in the same way at the same time.HE and Masson staining were used to observing the pathological structural changes and fibrosis of renal glomerular and tubulointerstitial kidney tissue in mice.And the expression of Shh,Smo,and Gli1 was detected by the immunofluorescence method.ResultsExperiment One The mice in the model group showed signs of apathy,decreased activity,reduced eating food and drinking water intake,displayed dull hair and partial hair shedding,and decreased urination and defecation.The vital signs of the mice in the control group were stable and in good condition.Through HE staining,we observed obvious pathological damage in the kidneys of the model mice,including renal tubular epithelial cells swelling with cytoplasmic vacuolation,a large number of inflammatory cells infiltration in the tubulointerstitium and around the blood vessels,glomerular mesangial proliferation,basement membrane hyperplasia,contrasting the normal renal morphology in control mice.Masson’s trichrome staining showed an obvious blue-positive stain in the glomerulus and renal tubules of the mice in the model group,indicating a higher collagen deposition level.Control mice had only a small amount of collagen deposition.By calculating the fibrosis area of the two groups,the statistical results showed that there was a significant difference between the two groups（P<0.01）.Experiment Two The results of Immunofluorescence showed that Shh,Smo and Gli1 proteins were significantly positively expressed in the renal tubules of mice in the model group,and compared with the control,the difference was statistically significant（P<0.01）.Experiment Three After moxibustion treatment,through HE staining and Masson staining,we found that the pathological changes in aged mice kidneys were reversed,including the reduction of inflammatory cell infiltration and collagen deposition,and so on.Compared with the model group,the renal fibrosis of the mice in the moxibustion group was remarkably reduced,and there was a significant difference between the two groups（P<0.01）.The immunofluorescence showed the expressions of Shh,Smo,and Gli1 at the protein levels in the moxibustion group were remarkably reduced,and compared with the model group,the difference was significant（P<0.01）.Conclusion10-month-old SAMP8 exhibited typical kidney fibrosis,and the high expression of Shh,Smo,and Gli1 indicated that the Sonic Hedgehog signaling pathway in aged mice kidneys was abnormally activated.Moxibustion at BL-23（Shen-shu）can markedly attenuate the fibrotic responses in the kidneys and evidently down-regulate the expressions of Shh,Smo,and Gli1 at the protein levels.Therefore,it is speculated that moxibustion at BL-23（Shen-shu）may repair kidney damage by inhibiting the abnormal activation of the Sonic Hedgehog signaling pathway. |
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