Yiqi Huoxue Tongluo recipe regulates NR4A1 to improve renal mitochondrial function in unilateral ureteral obstruction (UUO) rats.

CONTEXT: Yiqi Huoxue Tongluo recipe (YHTR) is a traditional Chinese medicine for the treatment of chronic kidney disease, but its exact mechanism is not clear. OBJECTIVES: To monitor the potential improvement of renal mitochondrial function in unilateral ureteral obstruction (UUO) rats by regulating NR4A1 using the YHTR. MATERIALS AND METHODS: Wistar rats were randomly divided into four groups: sham, UUO (left ureteral ligation for 14 days), eplerenone (EPL) (UUO + EPL), and YHTR (UUO + YHTR). UUO rats were established and intragastrically administered EPL (100 mg/day/kg) or YHTR (11.7 g/day/kg) for 14 days. The expression of related proteins in kidneys was detected by immunohistochemistry, western blot, RT-PCR, and chemical colorimetric assay, respectively. RESULTS: In vivo, YHTR treatment reduced the levels of BUN and Scr (by 17.9% and 23.5%) in UUO rats. Moreover, YHTR improved the renal mitochondrial function via increasing key enzymes of the tricarboxylic acid (TCA) cycle (p < 0.05) and activity of the mitochondrial complex (I-V) (by 30.8%, 29.1%, 19.7%, 35.9%, and 22.4%) in UUO rats. Compared with the UUO group, the expression of NR4A1 and Bcl-2 were significantly increased (p < 0.05), the expression of caspase-3 and caspase-9 were significantly decreased (p < 0.05) in the YHTR group. YHTR could upregulate key enzymes of the TCA cycle via promoting NR4A1 expression in HK2 cells, leading to inhibition of TGF-ß1 induced cell apoptosis. CONCLUSIONS: YHTR significantly improved the development of CKD; this study may provide new ideas for the pathogenesis of CKD and new strategies for the development of new drugs against CKD.

Huoxue Jiedu Huayu Formula Alleviates Cell Pyroptosis in Contralateral Kidneys of 6-Month-Old UUO Rats through the NLRP3/Caspase-1/IL-1ß Pathway.

OBJECTIVES: To study the protective effects and mechanisms of Huoxue Jiedu Huayu formula on cell pyroptosis through the NLRP3/caspase-1/IL-1ß pathway in contralateral kidneys in 6-month-old unilateral ureteral obstruction (UUO) rats. METHODS: Wistar rats were randomly assigned to 5 groups: a Sham group, a unilateral nephrectomy group (UNX group), a UUO group, a UUO treated with spironolactone group (Spi group), and a UUO treated with Huoxue Jiedu Huayu formula group (HJHF group). After 6 months of oral drug intervention, blood and contralateral kidneys were collected for research. RESULTS: The morphology and function of the contralateral kidneys were essentially normal after unilateral nephrectomy. HJHF obviously decreased serum creatinine, urea, and inflammatory lesions and depressed cell pyroptosis based on the NLRP3/caspase-1/IL-1ß pathway. Moreover, spironolactone, a mineralocorticoid receptor (MR) blocker, suppressed cell pyroptosis through SGK-1 and NF-кB. CONCLUSION: HJHF and spirolactone inhibited excessive activation of MR and then reduced cell pyroptosis, which was dependent on the NLRP3/caspase-1/IL-1ß pathway, to protect the contralateral kidneys of 6-month-old UUO rats.

Characterization of the complete chloroplast genome of Homalomena occulta (Lour.) Schott.

Homalomena occulta (Lour.) Schott (H. occulta) is a traditional Chinese medicine. However, the chloroplast genome has not been reported. Here, we assembled and analyzed the complete chloroplast (CP) genome of H. occulta. We found that the CP genome of H. occulta is 165,398 bp in length and contains a large single-copy (LSC) region of 92,861 bp, a small single-copy (SSC) region of 20,943 bp and an inverted repeat (IR) region of 25,797 bp. The genome contains 130 genes including 85 protein-coding genes, 8 rRNA and 37 tRNA. Phylogenetic analysis indicated that H. occulta is close to Philodendron lanceolatum. This study provides useful data for the development of molecular markers and identification of H. occulta.

Bushenhuoxue improves cognitive function and activates brain-derived neurotrophic factor-mediated signaling in a rat model of vascular dementia.

OBJECTIVE: To explore the protective mechanisms of the Traditional Chinese Medicine Bushenhuoxue (BSHX) in a rat model of vascular dementia (VD). METHODS: A rat model of VD was developed using bilateral common carotid artery occlusion (BCCAO). Rats were administered BSHX (10.14 or 5.07 g/kg), nimodipine (11.06 mg/kg; positive control), or saline (control) by gavage daily for 30 d post-surgery. Learning and memory abilities were assessed using the Morris water maze. Morphological changes in the hippocampus were observed using light microscopy (hematoxylin and eosin staining) and transmission electron microscopy (TEM). The mRNA and protein expression levels of brain-derived neurotrophic factor (BDNF), tyrosine receptor kinase B (TrkB), phosphatidyl inositol 3-kinase (PI3K), serine/threonine kinase (AKT), and cAMP response element binding protein (CREB) were measured by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: Compared with the sham group, rats with BCCAO exhibited impaired learning and memory abilities (Morris water maze) and showed abnormalities in neuronal morphology (light microscopy) and ultrastructure (TEM) in the hippocampus. They also had decreased mRNA and protein expressions of BDNF, TrkB, PI3K, AKT, and CREB in hippocampal tissue (all P < 0.05). In rats with BCCAO, administration of BSHX attenuated deficits in learning and memory, improved the morphology and ultrastructure of hippocampal neurons, and enhanced mRNA and protein expression levels of BDNF, TrkB, PI3K, AKT, and CREB (all P < 0.05). CONCLUSION: BSHX may protect hippocampal neurons and improve learning and memory abilities, at least in part via the activation of BDNF/TrkB/PI3K/AKT/CREB signaling.

Huoxue Jiedu Huayu Recipe Ameliorates Mesangial Cell Pyroptosis in Contralateral Kidney of UUO Rats.

OBJECTIVES: To observe the effects of the Huoxue Jiedu Huayu Recipe (HJHR) on pyroptosis of glomerular mesangial cells in the contralateral unobstructed kidney (CK) of unilateral ureteral obstruction (UUO) rats. METHODS: Sprague-Dawley rats were randomly divided into 4 groups: sham group, UUO group (10 days of left ureter ligation), UUO treated with eplerenone (EPL) (UUO + EPL) group, and UUO treated with HJHR (UUO + HJHR) group. The CKs of all rats were collected for studies. RESULTS: Cell pyroptosis and macrophage infiltration was found in contralateral glomeruli, and nucleotide-binding oligomerization domain-like pyrin domain containing protein 3 (NLRP3) and interleukin (IL)-1ß expression was upregulated in the CK of UUO rats. All of these changes were inhibited by HJHR and eplerenone. To determine how aldosterone (Aldo) activated the mineralocorticoid receptor (MR) and then induced mesangial cell pyroptosis with NLRP3-caspase-1-IL-1ß pathway, human mesangial cells (HMCs) were treated with HJHR and eplerenone, which were examined to detect the expression of NLRP3 inflammasome-associated proteins following treatment with Aldo. CONCLUSION: These results suggest that HJHR and eplerenone suppressed HMC pyroptosis via the MR/NLRP3 pathway.

[Effect of Huayu Jiedu Recipe on the Expressions of NLRP3, Caspasel, and IL-1 ß in Kidneys of Obstructive Nephropathy Rats].

Objective To observe the effect of Huayu Jiedu Recipe (HJR) on the expressions of nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) , Caspase-1 , IL-1 ß in kidneys of obstructive nephropathy rats. Methods Totally 40 clean grade SD rats were randomly divided into the sham-operation group (n =10) and the model group (n =30). The model of obstructive nephropa- thy was established by unilateral ureteral obstruction (UUO). Totally 30 successfully modeled UUO rats were randomly divided into the model group, the Western medicine group, the Chinese medicine group, 10 in each group. Eplerenone (100 mg . kg ⁻¹ . d⁻¹) was administrated to rats in the Western medicine group. HJR (13.7 g . kg ⁻¹ . d⁻¹) was administrated to rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medica- tion was performed once daily for 10 successive days. The serum IL-1 ß level was detected. Protein and mRNA expressions of NLRP3, Caspase-1, and IL-1 ß in renal tissue were detected. TUNEL positive rate was detected by TUNEL method. Results The expression of NLRP3 was not obviously seen, Caspase-1 and IL-1 ß were weakly expressed, and only fewer amount of TUNEL positive cells could be seen in the sham-operation group. Compared with the sham-operation group, serum IL-1ß level increased (P < 0. 01) , mRNA and protein expression of NLRP3, Caspase-1 , and IL-1 ß were up-regulated in renal tissue of the model group (P <0. 01). NLRP3 was mainly expressed in renal interstitial macrophages and renal tubular epithelial cells. Caspase-1 and IL-1 ß were mainly expressed in the cytoplasm of renal tubular epithelial cells. TUNEL positive cells were significantly increased, mainly dominated in interstitial expanded epithelial cells of distal tubules (P <0. 01). Compared with the model group, serum IL-1 ß level was significantly decreased (P <0. 01) ; mRNA and protein expressions of NLRP3, Caspase-1 , and IL- ß were obviously down-regulated (P <0. 01) , and the TUNEL positive rate was obviously decreased (P <0. 05, P < 0. 01) in the two medicated groups. Conclusion HJR could down-regulate mRNA and protein expres- sions of NLRP3, Caspase-1 , and IL-1ß, thus attenuating inflammatory injury of renal tissue.

[Protective effects of chailing decoction on cyclosporine A induced chronic renal injury and its mechanisms].

OBJECTIVE: To observe the effects of Chailing Decoction (CD) on transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), renal cell apoptosis and proliferation in rats with chronic cyclosporine A nephropathy (CCN), and to explore its possible mechanism for inhibiting renal fibrosis. METHODS: The CCN rat model was prepared using oral administration of cyclosporine A (CsA, 30 mg x kg(-1) x d(-1)). Meanwhile, they were treated with CD (3 g x kg(-1) x d(-1)) by gastrogavage. The serum blood urea nitrogen (BUN), serum creatinine (SCr), and creatinine clearance rate (CCr) were measured by the end of the fourth week of the experiment. The kidneys were taken out on the next day. The degree of renal fibrosis was detected using Masson staining. The protein and gene expressions of TGF-beta1, and CTGF were observed using immunohistochemical assay and RT-PCR. The renal cell apoptosis rate and the proliferation index were detected by flow cytometry. RESULTS: Compared with the control group (BUN: 6.123 +/- 0.588 mmol/L; SCr: 75.654 +/- 8.196 micromol/L; CCr: 0.539 +/- 0.169 mL/min), the renal function of the model group (BUN: 11.600 +/- 1.437 mmol/L; SCr: 101.985 +/- 10.809 micromol/L; CCr: 0.272 +/- 0.060 mL/min) obviously declined (P < 0.01). The collagen deposition in the renal interstitial area significantly increased. The protein and mRNA expressions of TGF-beta1, and CTGF in the tubular epithelial cells and the mesenchymal cells were significantly enhanced (P < 0.01). The cell proliferation index and the apoptosis rate both increased, but the ratio of apoptosis to proliferation (0.317 +/- 0.059) decreased more than that in the control group (0.680 +/- 0.150, P < 0.01). After treatment by CD, the renal function (BUN: 7.340 +/- 0.857; SCr: 84.923 +/- 10.627; CCr: 0.405 +/- 0.081) was significantly enhanced (P < 0.05, P < 0.01), the collagen deposition decreased, the high protein and mRNA expressions of TGF-beta1 and CTGF were down-regulated (P < 0.01), the ratio of apoptosis to proliferation increased (0.650 +/- 0.092, P<0. 01). CONCLUSION: CD could improve the renal function of CCN model rats, inhibit the expressions of TGF-beta1 and CTGF, and recover the balance between the renal cell apoptosis and proliferation by inducing cell apoptosis and inhibiting cell proliferation, thus delaying the renal fibrosis process.

[Inhibitory effect of chailing decoction on renal tubular epithelial phenotype transformation in rats with cyclosporine A-induced nephropathy].

OBJECTIVE: To observe the effect of Chailing Decoction (CLD) on alpha-smooth muscular actin (alpha-SMA, a marker of renal tubular epithelial phenotype transformation) in rats with cyclosporine A (CsA)-induced nephropathy for investigate its mechanism of action in inhibiting tubulo-interstitial fibrosis. METHODS: Forty Sprague-Dawley rats were equally randomized into four groups: the normal group, the model group, the positive control group and the Chinese medicine (CM) group. Excepting those in the normal group received gastrogavage with olive oil, rats in the other three groups were made into nephropathy model by oral infusion of CsA 30 mg/ (kg x d) for 28 days. At the same time of modeling, rats in the positive control and CM groups were treated respectively with Valsartan 10 mg/(kg x d) and CLD 3 g/(kg x d). The mRNA and protein expressions of alpha-SMA in rats' kidney were detected by RT-PCR, Western blot, immunohistochemical and flow cytometry, and the mRNA expressions of transforming growth factor-beta1 (TGF-beta1) and collagen type III (Col III) were determined by RT-PCR. RESULTS: Levels of (alpha-SMA, TGF-beta1, and Col III were significantly higher in the model group than those in the normal group respectively (P < 0.01 or P < 0.05), and the high levels were down-regulated in the positive control and CM groups after treatment (P < 0.01 or P < 0.05). CONCLUSION: CLD could retard the progress of renal interstitial fibrosis by way of inhibiting renal tubular epithelial phenotype transformation.