Effects of Hirsutella sinensis on TGF-beta1 and Snail expressions and transdifferentiation of tubular epithelial-myofibroblast in renal tissue of rats with chronic aristolochic acid nephropathy / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the antagonizing effect of Hirsutella sinensis (HS) on renal tubular epithelial-myofibroblast transdifferentiation (TEMT) and its possible pathogenic mechanism in rats with chronic aristolochic acid nephropathy (CAAN).</p><p><b>METHODS</b>Eighteen male Sprague-Dawley rats were equally divided into 3 groups, the model (M) group, the intervention (I) group and the control (C) group. The 24 h urinary protein (UP) in rats was measured before intervention and at the end of the 1st, 4th, 8th, and 12th week, and creatinine clearance rate (CCr) was measured before intervention and at the end of the 12th week respectively. All rats were sacrificed at the end of the 12th week, their kidney was taken for examining the degree of fibrosis in renal interstitial with Masson's stain and determining mRNA and protein expressions of transforming growth factor-beta1 (TGF-beta1), Snail, alpha-smooth muscle actin (alpha-SMA) and cytokeratin in renal tissue by Real-time RT-PCR and immunohistochemistry staining, respectively.</p><p><b>RESULTS</b>Compared with the C group, CCr was significantly lower, while 24 h UP was higher; the relative area of interstitial fibrosis was significantly larger in the M group; besides, the mRNA and protein expressions of TGF-beta1, Snail and alpha-SMA were significantly up-regulated (P < 0.01 or P < 0.05), and those of cytokeratin were significantly down-regulated (P < 0.01) in renal tissue of the M group. While in the I group, all the above-mentioned abnormalities were restored to some extent (P < 0.05) and showed significant difference (all P < 0.05) as compared with those in the M group.</p><p><b>CONCLUSION</b>HS can downregulate TGF-beta1 and Snail expressions in renal tissue, antagonize TEMT and renal interstitial fibrosis, and improve renal function in CAAN rats.</p>

The medicinal serum of yi-shen ruan-jian san antagonized the effect of aristolochic acid on human proximal tubular epithelial cells in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate whether the medicinal serum of Yi-shen Ruan-jian san can antagonize the fibrogenic effect of human proximal tubular epithelial cell line (HKC) activated by aristolochic acid (AA) in vitro.</p><p><b>METHOD</b>The HKC was incubated in the media containing 40 mg x L(-1) aristolochic acid sodium salt (AA-Na) with or without 10% concentration of Yi-shen Ruan-jian san medicinal serum. Then the cell proliferation and cytotoxicity of HKC were determined by MTF and lactate dehydrogenase (LDH) release assay respectively, the antigen expression of cytokeratin and alpha-smooth muscle actin on HKC was detected by immunocytochemistry, the mRNA expression of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), plasminogen activator inhibitor-1 (PAI-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and type I Collagen (Col I) of HKC was measured by RT-PCR, and their protein expression was measured by ELISA or Western blot.</p><p><b>RESULT</b>No cytotoxic effect was found in HKC after stimulation of AA-Na with or without the medicinal serum of Yi-shen Ruan-jian san (P > 0.05). No epithelial-myofibroblast transdifferentiation was found in HKC after AA-Na stimulation. The mRNA and protein expression of TGF-beta1, CTGF, PAI-1 and TIMP-1 of HKC was significantly upregulated by AA-Na (P < 0.05). The above-mentioned enhanced mRNA and protein expression, except for PAI-1, was significantly downregulated by the medicinal serum of Yi-shen Ruan-jian san, compared with the control (normal rat serum in the same concentration) (P < 0.05).</p><p><b>CONCLUSION</b>The fibrogenic effects of HKC activated by AA are antagonized by Yi-shen Ruan-jian san, through downregulating the expression of promoting excellular matrix (ECM) synthesis factors (TGF-beta1, CTGF) and inhibiting ECM degradation factor (TIMP-1).</p>

Protective effects of yishen ruanjian power on renal interstitial fribrosis in chronic aristolochic acid induced nephropathy rat model / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the protective effects of Yishen Ruanjian Power (YRP) on renal interstitial fibrosis in rats with chronic aristolochic acid induced nephropathy (CAAN).</p><p><b>METHODS</b>Eighteen male SD rats were divided into 3 groups, 6 in each group. Water solution of Caulis Aristolochia Manshuriensis (CAM) Liquid Extract were given to the mice in the model group by gastrogavage to make CAAN animal model. For those in the TCM group, decocted water solution of YRP was given by gastrogavage after the mice being modeled with the above-mentioned method. Tap water was given by gastrogavage to the mice in the control group. Body weight, 24-hr urinary protein excretion and beta2 microglobulin (beta2-MG), and serum creatinine (r) were determined at the end of the 1st, 4th, 8th, 12th and 16th week. At the end of the 16th week, the rats were sacrificed and the pathological figure of their kidneys were observed by Masson staining. Transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), plasminogen activator inhibitor-1 (PAI-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and type I collagen (Col I ) in kidney tissue were determined by RT-PCR and immunohistochemical method, respectively.</p><p><b>RESULTS</b>At end of the 1st week, urinary protein excretion, urinary beta2-MG and SCr in the model group were significantly increased to the levels higher than those in the control group (P < 0.01 or 0.05). Relative area of interstitial fibrosis was significantly enlarged in the model group at the end of the 16th week (P<0.01), and at the same time, the mRNA and protein expression of TCF-beta1, CTGF, PAI-1, TIMP-1 and Col I in kidney tissue were significant up-regulated (P<0.01). After intervention with YRP, the above-mentioned up-regulated parameters, except 24-hr urinary protein excretion, were all significantly inhibited (P <0.01 or 0.05).</p><p><b>CONCLUSION</b>YRP could inhibit the accumulation of extracellular matrix in renal interstitial tissue, so as to alleviate the renal interstitial fibrosis and improve the renal function.</p>

Antagonistic effect of yishen ruanjian san contained serum against aristolochic acid in antagonizing human renal interstitial fibroblasts / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study whether yishen ruanjian san contained serum (S-YRS) could intervene the action of aristolochic acid (AA) in antagonizing human renal interstitial fibroblasts (hRIFs) to induce extracellular matrix (ECM) accumulation.</p><p><b>METHODS</b>AA-Na 40 microg/ml, with or without 10% S-YRS, was co-cultured with hRIFs, then the hRIFs mRNA of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), plasminogen activator inhibitor-1 (PAI-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and type I collagen (Col I) in the cultured cells were detected by RT-PCR, and their protein expression monitored with ELISA and Western blot respectively.</p><p><b>RESULTS</b>The mRNA and protein expression of all the above-mentioned factors were significantly up-regulated by AA-Na (P < 0.05). Excepting PAI-1, the enhanced mRNA and protein expression were significantly down-regulated by S-YRS (P < 0.05).</p><p><b>CONCLUSION</b>S-YRS could down-regulate the hRIF to promote the expression of ECM synthesis factors and inhibit the ECM degradation factors in hRIFs, so as to antagonize the AA stimulated accumulation of ECM such as Col I.</p>

Multicenter prospective study on relationship of TCM syndrome type and renal pathology in 286 patients with IgA nephropathy / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the distribution pattern of TCM Syndrome type and its relationship with renal pathology in patients with IgA nephropathy.</p><p><b>METHODS</b>Apopting multicenter coordinated method, patients' TCM Syndrome type was differentiated according to their clinical manifestations, data concerning laboratory examination and renal pathology were collected to establish a database for analyzing the relationship between TCM Syndrome type and renal pathology in 286 patients.</p><p><b>RESULTS</b>Patients of Pi-Fei Qi-deficiency type (type 1) and both Qi-Yin deficiency type (type 2) showed rather milder pathological changes, by Lee classification, most of them belonged to grade I-III (72.3%, 70.2%); patients of Gan-Shen Yin deficiency type (type 3) had severe pathological change and majority of them belonged to grade III-IV (84.6%); and the most severe pathological change was shown in patients of Pi-Shen Yang-deficiency type (type 4), and the Lee's grade IV-V was dominant (88.0%) in them. Syndrome type of patients was significantly correlated with their Lee's grade (r = 0.26, P < 0.01). Percentage of glomerular sclerosis in patients of type 4 was higher than that in patients of the other three types. Semi-quantitative scoring upon pathological changes showed that the total, glomerular, tubulo-interstitial and vascular scores were significantly higher in patients of type 4 than in those of the other three types; those scores were higher in patients of type 3 than in type 1; the total, glomerular and vascular scores were higher in patients of type 3 than in type 2; and scores in patients of type 1 and type 2 showed insignificant difference.</p><p><b>CONCLUSION</b>Multicentric prospective study proves that the TCM Syndrome typeof patients with IgA nephropathy is significantly correlated with the grade and severity of their renal pathological changes, thus, the TCM Syndrome typing shows definite referential importance to conclude the severity of renal pathological change in patients with IgA nephropathy.</p>