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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 1-9, 2024.
Article in Chinese | WPRIM | ID: wpr-1006549

ABSTRACT

ObjectiveTo explore the mechanism of modified Liuwei Dihuangtang in preventing and treating renal injury in diabetic kidney disease (DKD) via the angiotensin-converting enzyme 1 (ACE1)/angiotensin Ⅱ (AngⅡ)/angiotensin Ⅱ type 1 receptor (AT1R) axis. MethodFifty male SD rats were randomized into a normal group (n=8) and a modeling group (n=42). The rats in the modeling group were fed with a high-sugar and high-fat diet for 6 weeks and intraperitoneally injected with 35 mg·kg-1 streptozotocin (STZ) to establish the model of DKD. After successful modeling, the rats were randomized into model, traditional Chinese medicine (modified Liuwei Dihuangtang granules 21 g·kg-1), western medicine (losartan potassium, 33 mg·kg-1), and integrated Chinese and western medicine (losartan potassium 33 mg·kg-1 combined with modified Liuwei Dihuangtang granules 21 g·kg-1) groups. The levels of fasting blood glucose (FBG), urinary protein (Up), blood urea nitrogen (Bun), and serum creatinine (SCr) were measured in each group after 8 consecutive weeks of drug intervention. Enzyme-linked immunosorbent assay was employed to determine the serum levels of ACE1, AngⅡ, and AT1R. Western blot was employed to measure the protein levels of ACE1, AngⅡ, and AT1R in the renal tissue. The pathological and morphological changes of the renal tissue were observed after hematoxylin-eosin (HE) staining, Masson staining, and periodic acid Schiff 's (PAS) staining. The fecal samples of rats in each group were collected for 16S rDNA high-throughput sequencing. ResultCompared with the normal group, the model group showed elevated levels of Up, FBG, Bun, SCr, ACE1, AngⅡ, and AT1R (P<0.01), serious lesions in the renal tissue, up-regulated protein levels of ACE1, AngⅡ, and AT1R (P<0.01), increased Firmicutes/Bacteroidetes (F/B) ratio, decreased relative abundance of Lactobacillus, and increased relative abundance of Moralella and Bifidobacteria. Compared with the model group, drug intervention lowered the levels of Bun, SCr, ACE1, AngⅡ, and AT1R (P<0.01) and alleviated the pathological changes in the renal tissue. Chinese medicine and integrated Chinese and western medicine lowered the levels of Up and FBG (P<0.01), and western medicine and integrated Chinese and western medicine down-regulated the protein levels of ACE1, AngⅡ, and AT1R. In addition, Chinese medicine down-regulated the protein levels of AngⅡ (P<0.01) as well as ACE1 and AT1R (P<0.05). Chinese medicine and integrated Chinese and western medicine decreased the F/B ratio, and western medicine and Chinese medicine increased the relative abundance of Blautia. Chinese medicine and integrated Chinese and western medicine increased the relative abundance of Lactobacillus, Ruminococcus undetermined genera, and Bifidobacteria, decreased the relative abundance of Moralella, and increased the Chao 1 and Ace indexes (P<0.05). Compared with the western medicine group, the integrated Chinese and western medicine group showed lowered levels of Up (P<0.01), Bun (P<0.05), and ACE1 and AT1R (P<0.01), down-regulated protein levels of ACE1, AngⅡ, and AT1R (P<0.05), alleviated pathological changes in the renal tissue, increased relative abundance of Bifidobacteria, and increased Chao 1 and Ace indexes (P<0.05). ConclusionModified Liuwei Dihuangtang combined with losartan potassium can mitigate renal fibrosis by regulating the ACE1/AngⅡ/AT1R axis, increasing the relative abundance of Lactobacillus and Bifidobacterium, reducing the relative abundance of Moralella, improving the richness and evenness of intestinal flora, and alleviating pathological damage in the renal tissue.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 61-70, 2021.
Article in Chinese | WPRIM | ID: wpr-905896

ABSTRACT

Objective:To observe the effects of modified Liuwei Dihuangtang on serum fibroblast growth factor 23 (FGF23), full-length intact parathyroid hormone (iPTH), and 1,25-dihydroxyvitamin D<sub>3 </sub>[1,25(OH)<sub>2</sub>D<sub>3</sub>] levels and Klotho and FGF23 protein expression in renal and bone tissues of rats exposed to high phosphorus combined with adenine, so as to explore the mechanism of modified Liuwei Dihuangtang against renal osteopathy. Method:One hundred and thirty healthy adult SD rats were randomly divided into five groups, namely normal group(<italic>n</italic>=10),high phosphorus group(<italic>n</italic>=30),model group(<italic>n</italic>=30),modified Liuwei Dihuangtang group(<italic>n</italic>=30) , and calcitriol group(<italic>n</italic>=30),and rats in each group were further classified based on three time points, namely 8,10, and 12 weeks. Rats in the normal group were fed with normal diet, the ones in the high phosphorus group with high phosphorus diet, and those in the other groups with adenine and high phosphorus diet for inducing renal osteopathy. Rats in the normal group,high phosphorus group, and model group were intragastrically administered with distilled water (10 mL·kg<sup>-1</sup>·d<sup>-1</sup>),the ones in the modified Liuwei Dihuangtang group with modified Liuwei Dihuangtang (2.556 g·kg<sup>-1</sup>·d<sup>-1</sup>) , and those in the calcitriol group with calcitriol (0.09 μg·kg<sup>-1</sup>·d<sup>-1</sup>). Result:Compared with the normal group and high phosphorus group at the weeks of 8,10 and 12,the model group displayed significantly elevated blood urea nitrogen(BUN),serum creatinine(SCr),serum phosphorus,iPTH,FGF23,renal interstitial fibrosis score, and FGF23 expression in renal and bone tissues, but lowered serum calcium and 1,25(OH)<sub>2</sub>D<sub>3</sub> and Klotho protein expression in renal and bone tissues(<italic>P</italic><0.05 ,<italic>P</italic><0.01). Compared with the model group at the weeks of 8,10 and 12, the modified Liuwei Dihuangtang and calcitriol both significantly decreased the serum BUN,SCr,serum phosphorus,iPTH, FGF23, tubulointerstitial semi-quantitative score, and FGF23 expression in renal and bone tissues, while increased the serum calcium,1,25(OH)<sub>2</sub>D<sub>3</sub>, and Klotho protein expression in renal and bone tissues (<italic>P</italic><0.05,<italic>P</italic><0.01). There was no significant difference in the above-mentioned indexes between the modified Liuwei Dihuangtang group and the calcitriol group at the same time point. Conclusion:Klotho-FGF23 axis is probably involved in renal osteopathy. The modified Liuwei Dihuangtang effectively improves renal function,alleviates pathological changes in renal and bone tissues,and regulates calcium and phosphorus metabolism to protect the bone, which is related to its regulation of Klotho-FGF23 axis.

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