The effects of Selenohomolanthionine supplementation on the rumen eukaryotic diversity of Shaanbei white cashmere wether goats.

Selenium (Se) is an important microelement for animal health. However, the knowledge about the effects of Se supplementation on rumen eukaryotic community remains less explored. In this study, the ruminal eukaryotic diversity in three months old Shaanbei white cashmere wether goats, with body weight (26.18 ± 2.71) kg, fed a basal diet [0.016 mg/kg Se dry matter (DM), control group (CG)] were compared to those animals given basal diet supplemented with different levels of organic Se in the form of Selenohomolanthionine (SeHLan), namely low Se group (LSE, 0.3 mg/kg DM), medium Se group (MSE, 0.6 mg/kg Se DM) and high Se group (HSE, 1.2 mg/kg DM) using 18S rRNA amplicon sequencing. Illumina sequencing generated 2,623,541 reads corresponding to 3123 operational taxonomic units (OTUs). Taxonomic analysis revealed that Eukaryota (77.95%) and Fungi (14.10%) were the dominant eukaryotic kingdom in all samples. The predominant rumen eukaryotic phylum was found to be Ciliophora (92.14%), while fungal phyla were dominated by Ascomycota (40.77%), Basidiomycota (23.77%), Mucoromycota (18.32%) and unidentified_Fungi (13.89%). The dominant eukaryotic genera were found to be Entodinium (55.44%), Ophryoscolex (10.51%) and Polyplastron (10.19%), while the fungal genera were dominanted by Mucor (15.39%), Pichia (9.88%), Aspergillu (8.24%), Malassezia (7.73%) and unidentified_Neocallimastigaceae (7.72%). The relative abundance of eukaryotic genera Ophryoscolex, Enoploplastron and fungal genus Mucor were found to differ significantly among the four treatment groups (P < 0.05). Moreover, Spearman correlation analysis revealed that the ciliate protozoa and fungi were negatively correlated with each other. The results of this study provided newer information about the effects of Se on rumen eukaryotic diversity patterns using 18s rRNA high-throughput sequencing technology.

Supplemental dietary Selenohomolanthionine affects growth and rumen bacterial population of Shaanbei white cashmere wether goats.

Selenium (Se) is an important trace element for all livestock growth. However, little is known about the dietary supplementation of Selenohomolanthionine (SeHLan) effect on growth and rumen microbiota of cashmere goats. In this study, thirty-two growing Shaanbei white cashmere wether goats with mean body weight (26.18 ± 2.71) kg were randomly assigned into 4 treatments, each with 8 replicates. The goats in 4 experimental groups were fed the basal diet (0.016 mg/kg Se) added with organic Se in the form of SeHLan, namely, control group (CG, added 0 mg/kg Se), low Se group (LSE, added 0.3 mg/kg Se), medium Se group (MSE, added 0.6 mg/kg Se), and high Se group (HSE, added 1.2 mg/kg Se). The feed experiment lasted for 70 days including 10-day adaptation, followed by 11 days digestibility trial including 7-day adaptation and 4-day collection period. On the last day of feeding experiment, rumen fluid was collected for microbial community analysis. The feed, orts, and fecal samples were collected for chemical analysis during digestibility trial. The results showed that average daily feed intake (ADFI) and the apparent digestibility of crude protein (CP) were both quadratic ally increased with increased SeHLan supply (P quadratic < 0.05), while average daily gain (ADG) and feed conversion ratio (FCR) showed a linear response (P linear < 0.05). The ADFI and ADG were all highest in the MSE group, which also had the lowest FCR (P < 0.05). Alpha diversity indices of the microbial community did not differ among four treatments. While principal coordinates analysis (PCoA) showed that rumen bacterial population differed among four groups. Taxonomic analysis revealed that Bacteroidetes, Firmicutes, and Euryarchaeota were the dominant phyla. The dominant families were Prevotellaceae, Selenomonadaceae, Methanobacteriaceae, and Bifidobacteriaceae. The significantly different rumen bacterial genera were found to be Methanobrevibacter, Quinella, Christensenellaceae_R-7_group, Veillonellaceae_UCG-001, and Succinivibrionaceae_UCG-002 (P < 0.05). In addition, Tax4fun analysis revealed that SeHLan supplemented groups enhanced the enrichment of genes related to energy metabolism, amino acid metabolism, carbohydrate metabolism, and enzymes. Twenty-eight pathways showed significant differences among four treatment groups (P < 0.05). In conclusion, dietary supplementation of medium SeHLan significantly affects rumen bacterial composition and ultimately promotes Shaanbei white cashmere wether goats nutrient digestibility and growth.

Single Cell RNA Sequencing Identifies a Unique Inflammatory Macrophage Subset as a Druggable Target for Alleviating Acute Kidney Injury.

Acute kidney injury (AKI) is a complex clinical disorder associated with poor outcomes. Targeted regulation of the degree of inflammation has been a potential strategy for AKI management. Macrophages are the main effector cells of kidney inflammation. However, macrophage heterogeneity in ischemia reperfusion injury induced AKI (IRI-AKI) remains unclear. Using single-cell RNA sequencing of the mononuclear phagocytic system in the murine IRI model, the authors demonstrate the complementary roles of kidney resident macrophages (KRMs) and monocyte-derived infiltrated macrophages (IMs) in modulating tissue inflammation and promoting tissue repair. A unique population of S100a9hi Ly6chi IMs is identified as an early responder to AKI, mediating the initiation and amplification of kidney inflammation. Kidney infiltration of S100A8/A9+ macrophages and the relevance of renal S100A8/A9 to tissue injury is confirmed in human AKI. Targeting the S100a8/a9 signaling with small-molecule inhibitors exhibits renal protective effects represented by improved renal function and reduced mortality in bilateral IRI model, and decreased inflammatory response, ameliorated kidney injury, and improved long-term outcome with decreased renal fibrosis in the unilateral IRI model. The findings support S100A8/A9 blockade as a feasible and clinically relevant therapy potentially waiting for translation in human AKI.

Analysis of Chuanxiong Rhizoma and its active components by Fourier transform infrared spectroscopy combined with two-dimensional correlation infrared spectroscopy.

As complicated mixture systems, active components of Chuanxiong Rhizoma are very difficult to identify and discriminate. In this paper, the macroscopic IR fingerprint method including Fourier transform infrared spectroscopy (FT-IR), the second derivative infrared spectroscopy (SD-IR) and two-dimensional correlation infrared spectroscopy (2DCOS-IR), was applied to study and identify Chuanxiong raw materials and its different segmented production of HPD-100 macroporous resin. Chuanxiong Rhizoma is rich in sucrose. In the FT-IR spectra, water eluate is more similar to sucrose than the powder and the decoction. Their second derivative spectra amplified the differences and revealed the potentially characteristic IR absorption bands and combined with the correlation coefficient, concluding that 50% ethanol eluate had more ligustilide than other eluates. Finally, it can be found from 2DCOS-IR spectra that proteins were extracted by ethanol from Chuanxiong decoction by HPD-100 macroporous resin. It was demonstrated that the above three-step infrared spectroscopy could be applicable for quick, non-destructive and effective analysis and identification of very complicated and similar mixture systems of traditional Chinese medicines.

[Modulation of aberrant extracellular matrix degradation systems by astragali radix and angelicae sinensis radix decoction (A&A) in interstitial fibrotic kidney].

OBJECTIVE: The imbalance between extracellular matrix (ECM) synthesis and degradation induces the excessive ECM deposition and thus renal fibrosis. The decoction (A&A) which is a combination of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has been shown to alleviate ECM production in animal models of chronic kidney diseases. In this paper, the effect of A&A on ECM degradation was investigated with interstitial fibrosis in rats. METHOD: Male Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After administration of A&A (14 g x kg(-1) x d(-1)) by gavage for 3, 7 and 10 days, morphological changes were evaluated by HE, PAS and Sirius red staining technique. The expression of plasminogen activator inhibitor-1 (PAI-1) and tissue-type plasminogen activator (t-PA), the activity of PAI-1 and t-PA were determined by ELISA. The activity of matrix metalloproteinases (MMP-9, MMP-2), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were evaluated by gelatin zymography or reverse gelatin zymography, respectively. RESULT: Morphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which was reduced by A&A administration (P < 0.05, UAA vs UUO group). Compared with the sham group, the expression of PAI-1 was significantly increased in UUO group by 63%, 91% and 112% at day 3, 7 and 10 respectively; and there were a remarkable decrease in UAA group by 44%, 43% and 52% at day 3, 7 and 10. The expression of active PAI-1 was strikingly increased in UUO group at day 3 [(30.5 +/- 23.8) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], day 7 [(36.5 +/- 11.2) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], and day 10 [(54.5 +/- 14.2) ng x g(-1) vs. (0.5 +/- 0.5) ng x g(-1), P < 0.05)]. The active PAI-1 was decreased in UAA group at day 7 [(14.9 +/- 0.5) ng x g(-1) vs. (36.5 +/- 11.2) ng x g(-1), P < 0.05] and day 10 [(15.4 +/- 4.8) ng x g(-1) vs. (54.5 +/- 14.2) ng x g(-1), P < 0.05]. The expression of t-PA was increased in UUO group only at day 3 [(58.1 +/- 16.5) microg x g(-1) vs. (30.1 +/- 17.3) microg x g(-1)], P < 0.05), meanwhile decreased in UAA group [(26.3 +/- 8.7) microg x g(-1) vs. (58.1 +/- 16.5) microg x g(-1), P < 0.05)]. But the expression of active t-PA was shown no significantly difference among the three groups. For MMP-2 and MMP-9 activity, they were significantly higher compared with the sham group in UUO group, but no significantly change after A&A treatment. The TIMP-1 activity was significantly increased in UUO group by 28% and 63% at day 7 and 10 respectively, significantly decreased in UAA group by 40% and 39% at the same time point. CONCLUSION: The anti-fibrosis effects of A&A might be associated with modulating the imbalance of PAs/PAIs system as well as MMPs/TIMPs system, thereby alleviate ECM accumulation and interstitial fibrosis.

Effects of Huanshuai Recipe Oral Liquid on restructuring glomerular microvasculature and expression of vascular endothelial growth factor in subtotal nephrectomized rats.

OBJECTIVES: To explore the effects and significance of Huanshuai Recipe Oral Liquid (, HSR), a formula with supplementing qi, nourishing blood and activating blood on restructuring glomerular microvasculature and expression of vascular endothelial growth factor (VEGF) in subtotal nephrectomized (SNX) rats. METHODS: A total of 76 male Wistar rats were randomly divided into four groups: 16 in the sham-operated group and fed with tap water 10 mL/kg per day; 20 in the model group were operated with 5/6 SNX and fed with tap water 10 mL/ kg per day; 20 SNX rats in the HSR group were treated with HSR 10 mL/kg per day; 20 SNX rats in the losartan group were treated with losartan 40 mg/kg per day. Serum creatinine (SCr) and urinary protein excretion (Upro) were examined at the 2nd, 4th, 8th, and 12th weeks of the treatment, and the remnant kidneys were harvested. Changes in histological microstructure were evaluated using light microscopy, and the expression of VEGF was detected by using ELISA. RESULTS: Upro, microvasculature injury and glomerulosclerosis were found to be alleviated in HSR and Losartan groups, respectively. The change of VEGF expression showed positive correlation with glomerular capillary area and peritubular capillary number (r=0.448, r=0.422, P<0.01), but negative correlation with that of SCr and Upro (r=-0.592, r=-0.481, P<0.01). CONCLUSIONS: HSR could regulate the VEGF expression, reduce the loss of microvasculature, which demonstrated similar renal protective effects to losartan in SNX rats. Examination of Chinese herbal medicine influence on VEGF signaling and restructuring renal microvasculature may elucidate the molecular mechanism of renal protection to a certain degree.

Altered expression of genes profiles modulated by a combination of Astragali Radix and Angelicae Sinensis Radix in obstructed rat kidney.

The decoction of a combination of two Chinese herbs, Astragali Radix (the roots of Astragalus membranaceus var. mongholicus) and Angelicae Sinensis Radix (the roots of Angelica sinensis), here named as A&A, has been demonstrated to have renoprotective effects in several animal models and may be considered as a complementary therapeutic medicine for chronic kidney disease. In this study, genomic approaches were employed to identify expression signatures in the obstructed kidney, which may be linked to the molecular actions associated with anti-fibrotic effects of A&A. Ninety-six male Wistar rats were divided randomly into sham, SAA (sham + A&A), UUO (unilateral ureteral obstruction), and UAA (UUO + A&A) groups. The rats in the SAA and UAA groups were administered A&A (14 g/kg) by oral gavage once daily; the ones in the sham and UUO groups were given equal volumes of water. Eight rats from each group were sacrificed at days 3, 7, and 10 after the operation, respectively. Changes in gene expression in the kidneys were determined using Affymetrix RAE-230A GeneChips. The differential expression of known genes between UAA and UUO was confirmed by RT-PCR. The results revealed that 40, 65, and 104 genes were upregulated and 30, 36, and 40 genes downregulated in UUO compared with the sham group at days 3, 7, and 10, respectively. Compared to the UUO group, eight genes were upregulated and two genes were downregulated at day 3 in the UAA group, and two genes were upregulated at day 10. These genes included transient receptor protein 3 (TRP3), bone marrow stromal cell antigen 1 (BST-1), peroxisomal biogenesis factor 6 (PEX6), xanthine dehydrogenase (XDH), cytochrome P450 subfamily I member A1 (CYP1A1), serine/cysteine proteinase inhibitor clade E member1 (PAI-1), fibroblast growth factor 23 (FGF23), and five ESTs. Among these genes, differential expression of PAI-1, FGF23, and CYP1A1 were further confirmed by RT-PCR. These data provide the evidence that the anti-fibrotic effects of A&A are mediated through multiple pathways in obstructive nephropathy, and novel mechanisms may be involved in the increasing degeneration of ECM, decreasing ROS reaction, and regulation of the calcium-phosphate metabolism.

[Screening effective parts of combination of Astragali Radix and Angelicae Sinensis Radix (A&A) for its renoprotective effects].

OBJECTIVE: The decoction of Astragali Radix and Angelicae Sinensis Radix (A&A) has shown antifibrotic effects in rats with unilateral ureteral obstruction (UUO). The aim of this study was to track the effective parts of A&A for its renoprotective effects, according to the improvement of renal function and renal tubulointerstitial damage. METHOD: A&A was sequentially extracted by using different solvents for three times and eleven different parts were gained. Wistar rats were randomly divided into Sham, UUO and the treatment groups with A&A or each part of A&A. After administration of A&A or its parts for 10 days, the levels of serum creatinin (Scr) and urea were measured. The morphological changes of kidneys were also semi-quantitatively analyzed by HE, Masson stained tissue sections, which including interstitial cell infiltration, tubular atrophy and interstitial fibrosis. RESULT: The levels of Scr, urea were significantly increased, accompanied with severe renal damage in rats with UUO. As same as A&A, the part I in the first extraction and part IC in the second extraction were all shown to decrease the levels of Scr and urea and the index of renal interstitial damage. However, the following 4 parts extracted from IC in the third extraction were shown no effect on the above indexes. CONCLUSION: The extract part I and part IC could be considered as the predominant parts of A&A for its renoprotective effects, due to their improvement of renal damage in interstitial nephropathy.

A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, improved renal microvascular insufficiency in 5/6 nephrectomized rats.

Chronic renal ischemia and hypoxia in the tubulointerstitium are involved in the mechanisms of progressive chronic kidney disease. Previous studies showed that the decoction of a combination of two Chinese herbs, Astragalus membmnaceus var. mongholicus and Angelica sinensis (A & A) has antifibrotic effects through multiple pathways in different animal models. In this study, remnant kidney model was employed to investigate whether A & A affect the expression of VEGF, the density of the renal microvasculature and thus alleviate the renal injury. Rats were divided randomly into four groups: sham group (N-31), 5/6 Nx group (5/ 6 nephrectomy, N=43), A & A treated group (A & A group, N=40, A & A 12 g/kg/d po), enalapril treated group (Ena group, N=56, enalapril 4 mg/kg/d po). Rats from each group were sacrificed at the 2th, 4th, 8th and 12th weeks respectively after surgery and treatment The 24 h urinary protein excretion, serum creatinine (Scr) and urea were measured. The collagen IV (COL-IV), fibronectin (FN), aminopeptidase P (APP) and VEGF were stained using immunohistochemistry. The COL-IV, FN and APP were semi-quantitatively analyzed. Peritubular capillary density in the cortical interstitial area was quantified. The level of VEGF was assayed by ELISA. The results revealed that Scr, urea and urinary protein excretion remained constant at each time point in sham group. Compared to sham group, 5/6 Nx group was shown severe glomerulosclerosis, tubulointerstitial lesions and vascular damage, as well as higher level of Scr from the 2nd week (72.3 +/- 5.2 vs. 48.6 +/- 2.6 micromol/L P < 0.05) to the 12th week (71.9 +/- 8.0 vs. 55.7 +/- 4.5 micromol/L P < 0.05). Although there was no significant difference in Scr level after treatment of enalapril or A & A (P > 0.05), kidney damage was alleviated at the 8th and the 12th week in the two treatment groups (P < 0.05, vs. 5/6 Nx group). The urinary protein excretion of 5/6 Nx group was significantly increased from the 4th week, it was 1.5, 2.4 and 3.8 fold of that of sham group at the 4th, 8th and 12th week, respectively. Compared to 5/6 Nx group, proteinuria was decreased by enalapril to 59%, 33% at 8th and 12th week. After A & A administration, urinary protein excretion decreased to 66%, 56%, 75%, 55% of 5/6 Nx group at the 2nd, 4th, 8th and 12th, respectively (P < 0.05). Compared with sham group, there was increased expression of FN and COL-IV in rats with 5/6 Nx. After A & A or enalapril administration, the expression of FN and COL-IV was significantly decreased compared with that in the 5/6 Nx group at 8th and 12th week (P < 0.05). On the other hand, the capillary density was decreased at the 8th and 12th week in 5/6 Nx rats (P < 0.01). In A & A-treated group, similarly with enalapril group, the amount of APP-positive glomerular capillary increased by 36% (P < 0.01), and the peritubular capillary density was increased 94% at 8th week and 52% at 12th week compared with 5/6 Nx group (P < 0.05). The renal level of VEGF was decreased in 5/6 Nx rats, but increased at the 8th and 12th week in A & A group (P < 0.05, vs. 5/6 Nx group). In conclusion, A & A has renoprotective effects by suppression of extra cellular matrix deposition in 5/6 Nx rat. The renoprotective effects may be associated with reduction of proteinuria, up-regulation of VEGF which may reduce the loss of capillaries and improve microstructure dysfunction.

[Renoprotective effects of Astragalus and Angelica mixture in rats with 5/6 nephrectomy].

OBJECTIVE: To explore whether Chinese herbs Astragalus and Angelica mixture (A&A) have renoprotective effects on chronic renal failure in rats. METHODS: 5/6 nephrectomized kidney in rats was used as chronic renal failure model. Rats were randomly divided into four groups which included sham, Model, A&A and enalapril (Ena) groups. The remnant kidney was harvested after 2, 4, 8, 12 weeks, and levels of serum creatinine (scr), urea and urinary protein excretion were measured using routine biochemical methods. The pathological lesions were observed by light microscope. The expressions of fibronectin (FN) and collagen IV (Col-IV) in glomeruli and interstitium were detected by immnuohistochemistry. RESULTS: 5/6 nephrectomized rats were presented with the characteristics of increasing scr and urea, persistent proteinuria at the end of the 2nd, 4th, 8th, 12th weeks, respectively. They had increasing size of glomeruli with progressive proliferation of mesangial cells and mesangial matrix followed by glomerulosclerosis and progressive elevated expressions of FN and Col-IV. Compared with Model group, the administration of A&A could decrease urinary protein excretion for 34%, 44%, 25%, 45% at the end of the 2nd, 4th, 8th and 12th weeks, respectively (P<0.05). At the end of the 8 th week of A&A administration, mesangial proliferation was much lower than that in the untreated rats, the expressions of FN and Col-IV were decreased for 18% and 29%(P<0.05), respectively. At the end of the 12th week, the number of sclerotic glomeruli was decreased and renal interstitial fibrosis almost disappeared, accompanied with the decreased expressions of FN and Col-IV for 17% and 14% (P<0.05), respectively. The beneficial effects of A&A were similar to the effects of enalapril in the study. CONCLUSION: A&A reduced extracellular matrix deposition and down-regulated the lesions of glomerulosclerosis and renal interstitial fibrosis in rats with chronic renal failure, which may be due to its effect on the earlier and persistent reduction of proteinuria.

A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, enhanced nitric oxide production in obstructed rat kidney.

BACKGROUND: The persistent renal hemodynamic maladjustment caused by imbalances between vasoactivators predisposes the kidney to tubulointerstitial injury and ultimate interstitial fibrosis. The decoction (A&A) of a combination of roots of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has shown antifibrotic effects in rats with chronic kidney diseases and improvement of renal blood flow in rats with acute ischemic renal injury. In the present study, we investigated the effects and possible mechanisms of A&A on vasoactivators in the process of renal interstitial fibrosis. METHODS: Male Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After oral administration of A&A (14 g/kg/d) for 3, 7 and 10 days, morphological changes were evaluated by HE, Masson and Sirius red staining technique. The levels of Ang-II, ET-1, and the activities of different nitric oxide synthases (NOSs) in renal homogenate were measured by radioimmunoassay. The nitrite concentration as nitric oxide (NO) production was measured using the Griess reagent. Western blot analysis and immunohistochemical staining were performed to determine the expressions of eNOS, nNOS, and iNOS in the kidney. The ability of scavenging reactive oxygen species (ROS) was evaluated by spectrophotometry. RESULTS: Morphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which reduced by A&A administration (p<0.05, UAA vs. UUO group). The levels of Ang-II and ET-I were increased in obstructed kidneys, but not significantly changed after A&A administration. NO production did not change in obstructed kidney at day 3 but increased in day 7 and day 10. Administering A&A progressively increased NO production by 2.2, 1.2, and 1.2 fold at days 3, 7 and 10, respectively. The activities of constitutive NOS and iNOS were comparable between UUO group and sham group. In contrast, the activity of constitutive NOS was much higher in UAA than that of UUO rats, which increased 78%, 68% and 78% at days 3, 7 and 10 respectively, although the protein expression of eNOS, nNOS and iNOS in renal tissue had no change in UAA rats. The activities of scavenging ROS in UUO group were not significantly different from the sham group at days 3 and 7, but increased at day 10 (24.1+/-15.0 vs. 10.1+/-0.8 U/min/mg protein, p<0.05). After A&A administration, the activities of scavenging ROS were significantly increased at days 3 and 7 (51.5+/-17.9 vs. 11.7+/-7.4 U/min/mg protein, p<0.05; and 16.1+/-5.6 vs. 7.7+/-1.4 U/min/mg protein, p<0.05) respectively, comparing with the UUO group. CONCLUSION: The anti-fibrosis effects of A&A might be associated with enhancing NO production via eNOS activation and scavenging ROS, and in turn might improve ischemic microvasculature and attenuate interstitial fibrosis.

[Studies on solid phase extraction method of aristolochic acids and aristololactams in rat urine].

OBJECTIVE: To develop a urine pretreatment method of Solid Phase Extraction (SPE) for the quantitative determination of a number of aristolochic acids (AAs) and aristololactams (ALs) in rat urine. METHOD: The HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other sixteen AAs and ALs was chosen as evaluating index to study the extract results of five Solid Phase Extraction columns (Agilent C18/100 mg, Alltech HG18/100 mg, Alltech C18/100 mg, Alltech C18/300 mg and Agilent Phenyl/200 mg) comparatively. The influences of two washing solvents (water and 1% acetic acid-0.02% triethylamine solution) and seven eluting solvents (ether, acetone, chloroform, ethyl acetate, dichloromethane, methanol and acetonitrile) on extract results of AAs and ALs are comparatively studied with the extracting recoveries of AA-I , AA-II, AL-I and AL-II as indicators. The HPLC peak area of AA-I , AA-II, AL-I and AL-II, and other seven AAs and ALs with good separation being targets, several factors which affect extracting efficiency of analytes, including activating volume, cleansing volume, washing volume and eluting volume, are optimized by orthogonal design experiments with four factors at three levels. RESULT: The established method of SPE is as follows: Agilent Phenyl SPE column of 200 mg, activating with 1.0 mL methanol, cleansing with 1 mL water, adding 1.0 mL rat urine sample, washing with 0.8 mL 1% acetic acid 0.02% triethylamine solution, and eluting with 3.0 mL methanol. CONCLUSION: The established method of SPE is efficient, selective, simple and fast, and can be used as urine pretreatment method to analyze a variety of aristolochic acids and aristololactams in rat urine.

[Studies on pharmacodynamic characteristics of aristolochic acid I in rats].

OBJECTIVE: To study pharmacodynamic characteristics by oral administration aristolochic acid I (AA-I) in rats. METHOD: After one-time oral administration of Aristolochiae manshuriensis decoction 10 g x kg(-1) and 125I labeled AA-I (containing AA-I 37.2 microg x mL(-1)), whole blood concentration of 125I-AA-I and the binding rate of serum albumin were detected in 69 normal wistar male rats. Metabolic dynamic parameters were calculated by program 3P87 with a two compartment model. The distribution ratio and ID% of nine viscera or tissue were measured and compared with other until the 40th day. RESULT: After oral administration, AA-I was rapidly absorbed into the blood and reached its peak at 30 minutes and lasted till 90 minutes. AA-I concentration in the blood gradually declined afterwards. 24 hours later, only few AA-I could be detected. By the 10th day, 68.5% of AA-I presented as the binding type with serum albumin. Pharmacodynamic parameters were calculated as follows: Tmax 0.74 h, Cmax 0.92 microg x mL(-1), t1/2alpha 0.68 h, t1/2beta 20.46 h, V/F 87.39 mL, CL(s) 5.85 mL x h(-1) (0.10 mL x min(-1)). On the other hand, after oral administration AA-I was rapidly distributed to all the viscera or tissue, whose peak appeared in 5 minutes and the vallecula was from 24 to 48 hours. The distribution ratio of AA-I rose in the kidney after 24 hours, and it showed the highest level in the kidney and in the liver by the 4th day compared with other organs or tissue (P < 0.05). However, the distribution ratio of AA-I in the kidney became the most dominant one after the 30th and the 40th day compared with the others (P < 0.05). CONCLUSION: AA-I is rapidly absorbed after oral administration in rats. Its distribution has the organ specificity, which is characterized as the possible partial metabolism in the liver and the accumulation in the kidney because of rather slower elimination. The characteristics may be related to the long term nephrotoxicity of AA-I.

[Preventive and therapeutic effects of astragalus and angelica mixture on renal tubulointerstitial fibrosis after unilateral ureteral obstruction in rats].

OBJECTIVE: To investigate the preventive and therapeutic effects of Astragalus and Angelica mixture A&A on renal tubulointerstitial fibrosis after unilateral ureteral obstruction UUO in rats and their mechanisms. METHODS: UUO rats were randomly divided into Sham, UUO, A&A or ACEI groups. A&A, ACEI or the same amount of water was administered by gavage beginning 24 hours before UUO preparation and continued through ten days after UUO. Sera and the kidney tissues were collected from each group on the tenth day. Scr and BUN were measured. Trichrome staining, measurement of tubulo interstitial damage index and immunohistochemical studies localizing alpha-smooth muscle actin alpha-SMA , TGF-beta1, fibronectin FN , laminin LN were carried out. RESULTS: In UUO rats, the tubular-interstitial damage index, the expressions of alpha-SMA, TGF-beta1, FN and LN were all increased compared with those of Sham group. The tubulo interstitial damage index had positive correlation with expressions of alpha-SMA, TGF-beta1, FN and LN. A&A significantly ameliorated deterioration of renal function, tubulo interstitial damage index and inhibited the over-expressions of alpha-SMA, TGF-beta1, FN and LN in UUO rats. These anti-fibrotic effects were similar to those affected by ACEI. CONCLUSION: In renal interstitial fibrosis-induced UUO rats, A&A retard the progression of renal fibrosis and renal function deterioration by inhibiting myofibroblasts and suppressing TGF-beta1 expression, which may consequently result in a decreased production of extracellular matrix.