KCNQ1OT1 Influences HK-2 Apoptosis and Inflammation in LPS-Induced Acute Renal Injury via Modulating miR-30a-5p/NLRP3 Axis.

Objective: To investigate the influence of KCNQ1OT1 on HK-2 apoptosis and inflammation in ARI and its molecular mechanism. Methods: Normal cultivated HK-2 cells were used as negative control (NC) group. Three different concentrations of lipopolysaccharide (LPS) were used to treat the cells (5 µg/mL, 10 µg/mL, and 20 µg/mL). The groups included si-KCN1OT1+ LPS, si-NC + LPS, miR-30a-5p + LPS, pcDNA-NLRP3+si-KCNQ1OT1 + LPS group, miR-NC + LPS group, and pcDNA + si-KCNQ1OT1 + LPS group. CCK-8 and flow cytometry are used to measure cell viability and apoptosis, while RT-qPCR and Western blotting are used to detect KCNQ1OT1, miR-30a-5p, and NLRP3 mRNA. ELISA was used to detect the levels of TNF-α, IL-6, and IL-1ß in HK-2 cells. The targeting relationship among KCNQ1OT1, miR-30a-5p, and NLRP3 was verified. Results: After the intervention of LPS, the viability of HK-2 cells was decreased, while the apoptosis rates were increased. The mRNA and protein expressions of NLRP3 and KCNQ1OT1 were increased, while the mRNA and protein levels of miR-30a-5p were decreased (P < 0.05). The expressions of Bax and Cleaved-caspase-3 were downregulated after silencing KCNQ1OT1 and overexpressed miR-30a-5p. In addition, the viability of HK-2 cells was improved, and the apoptosis was reduced by inhibiting KCNQ1OT1 and overexpressed miR-30a-5p. Thus, KCNQ1OT1 modulated NLRP3 via targeting miR-30a-5p. Overexpression of NLRP3 reverses KCNQ1OT1 inhibition of LPS-induced apoptosis, activity, and inflammation in HK-2 cells. Conclusions: Through modulating the miR-30a-5p/NLRP3 axis, inhibition of KCNQ1OT1 may reduce HK-2 apoptosis and inflammation in LPS-induced ARI.

Astragalus mongholicus Bunge and Panax notoginseng (Burkill) F.H. Chen Formula for Renal Injury in Diabetic Nephropathy-In Vivo and In Vitro Evidence for Autophagy Regulation.

BACKGROUND: Diabetic nephropathy (DN) is a serious complication of diabetes mellitus (DM) with limited treatment options. DN leads to progressive renal failure and accelerates rapidly into end-stage renal disease. Astragalus mongholicus Bunge and Panax notoginseng (Burkill) F.H. Chen formula (APF) is a traditional Chinese medicine (TCM) formula widely used to treat chronic kidney diseases (CKD) in the clinic in the southwest of China. The aim of this study is to explore how APF and its related TCM theory work on DN and whether mTOR/PINK1/Parkin signaling plays a part in this process. METHODS: HPLC was used for preliminary chemical analysis and quantitative analysis of the five components of APF. An in vivo autophagy deficiency model was established in C57BL/6 mice by streptozocin (STZ) combined with a high-fat and high-sugar diet, while the in vitro autophagy deficiency model was induced with high glucose (HG) in renal mesangial cells (RMCs). Renal histopathology staining was performed to investigate the extents of inflammation and injury. Real time-PCR and Western blotting techniques were utilized to assess autophagy-related proteins. RESULTS: APF significantly ameliorated renal injury in DN mice, specifically restoring blood urea nitrogen, serum creatinine, and 24-hour albuminuria. APF also reduced the mRNA and protein expressions of TNFα, IL-1ß, and IL-6 in STZ-induced DN mice. Furthermore, APF improved the autophagy deficiency induced by STZ in vivo or HG in vitro, as revealed by changes in the expressions of mTOR, PINK1, Parkin, Beclin 1, p62, and LC3B. Notably, inhibition of autophagy with 3-methyladenine in APF-treated RMCs aggravated cellular damage and altered mTOR/PINK1/Parkin signaling, indicating that APF rescued HG damage through promoting autophagy. CONCLUSION: APF may protect the kidneys from inflammation injuries in DN by upregulating autophagy via suppressing mTOR and activating PINK1/Parkin signaling. This experimental evidence strongly supports APF as a potential option for the prevention and treatment of DN.

[Effects of fast-twisting long-retaining acupuncture therapy on apoptosis and expression of related proteins in vestibular nucleus in rats with vertigo induced by posterior circulation ischemia].

OBJECTIVE: To observe the effects of fast-twisting long-retaining (FTLR) acupuncture therapy on apoptosis of vestibular nucleus and expression of Caspase-3, Bcl-2 and Bax in rats with vertigo induced by posterior circulation ischemia. METHODS: A total of 70 healthy SD rats were randomly divided into a sham operation group, a model group, a medication group, a regular acupuncture group and a FTLR acupuncture group, 14 rats in each group. The rats in the model group, medication group, regular acupuncture group and FTLR acupuncture group were intervented with surgical ligation of the right common carotid artery (CCA) and the right subclavian artery (SCA) to establish the model of vertigo induced by posterior circulation ischemia; in the sham operation group, the right CCA and the right SCA were separated without ligation. The rats in the medication group were treated with gavage of flunarizine hydrochloride suspension (10 mL/kg). "Baihui" (GV 20), "Shuaigu" (GB 8) and "Fengchi" (GB 20) were selected in the two acupuncture groups. The rats in the regular acupuncture group were treated with routine acupuncture and the needles were retained for 30 min, while the rats in the FTLR acupuncture group were treated with quick twist (200-300 times/min) for 1 min and the needles were retained for 60 min. The rats in the sham operation group and the model group received no intervention. All the intervention was provided once a day for 10 days. The decline rate of local blood flow in vestibular nucleus was observed; the apoptosis of vestibular nucleus was observed by TUNEL method; the expression of Caspase-3, Bcl-2 and Bax proteins were detected by immunohistochemistry. RESULTS: Compared with the sham operation group, the decline rate of local blood flow in the right vestibular nucleus was significantly increased in the model group (P<0.01), and the apoptosis index (AI) of vestibular nucleus was significantly increased (P<0.01). Compared with the model group, the decline rates of local blood flow in the right vestibular nucleus in the two acupuncture groups and medication group were significantly reduced (P<0.01), and the AIs of vestibular nucleus cells were significantly reduced (P<0.01). The decline rate of local blood flow in the right vestibular nucleus in the FTLR acupuncture group was lower than those in the medication group and the regular acupuncture group (P<0.01, P<0.05), and the AI of vestibular nucleus was lower than those in the regular acupuncture group and the medication group (P<0.05). Compared with the sham operation group, the expression of Bcl-2 in the vestibular nucleus was significantly decreased in the model group (P<0.01), and the expressions of Bax and Caspase-3 were significantly increased (P<0.01). Compared with the model group, the expressions of Bcl-2 in the vestibular nucleus were significantly increased in the two acupuncture groups and medication group (P<0.01), and the expressions of Bax and Caspase-3 were significantly reduced (P<0.01). The expression of Bcl-2 in the vestibular nucleus in the FTLR acupuncture group was higher than those in the regular acupuncture group and the medication group (P<0.05), and the expressions of Bax and Caspase-3 were lower than those in the regular acupuncture group and the medication group (P<0.05). CONCLUSION: The FTLR acupuncture therapy could effectively inhibit the apoptosis of vestibular nucleus in rats with vertigo induced by posterior circulation ischemia, and its mechanism may be related to improving the blood supply of vestibular nucleus and regulating the expressions of Caspase-3, Bcl-2 and Bax proteins.

[Treatment of Level 2 Hypertension by Diagnosis and Treatment Program of Integrative Medicine: a Multi-centre, Randomized Controlled Trial].

OBJECTIVE: To explore the therapeutic effect of diagnosis and treatment program of integrative medicine (IM) on level 2 hypertension in the young and middle-aged patients and their ambulatory blood pressure. METHODS: A randomized, placebo parallel and controlled, multi-center clinical trial was performed. Totally 199 young and middle-aged level 2 hypertension patients were randomly assigned to the treatment group (99 cases) and the control group (100 cases). All received combined hypotensive treatment program by taking Nifedipine Sustained Release Tablet and Hydrochlorothiazide as basic drugs. Patients in the treatment group additionally took Western medicine (WM) combined Jiangyabao serial drugs (0.31 g per tablet, 2 tablets each time, twice daily), while those in the control group additionally took WM combined simulative agents of Jiangyabao serial drugs (0.31 g per tablet, 2 tablets each time, twice daily). The treatment course was 8 weeks for all, and 24-week follow-ups performed. 24 h ambulatory blood pressure and casual blood pressure, and their efficacies were compared between the two groups, and safety assessed as well. RESULTS: Compared with before treatment in the same group, daytime and night casual blood pressure, as well as 24 h ambulatory blood pressure were all obviously improved in the two groups (P < 0.01). Average diastolic and systolic blood pressures at night decreased more in the treatment group than in the control group with statistical difference (P < 0.05). There was no statistical difference in total efficacies of daytime casual blood pressure or ambulatory blood pressure (P > 0.05). CONCLUSION: Jiangyabao serial drugs combined WM in treating young and middle-aged level 2 hypertension patients showed obvious effect in improving night blood pressure, especially for night diastolic blood pressure.

Ginsenoside Rg1 reduces aldosterone-induced autophagy via the AMPK/mTOR pathway in NRK-52E cells.

Aldosterone is a steroid hormone secreted from the adrenal cortex, which regulates blood pressure. Higher concentrations of aldosterone can cause several diseases, including hypertension, diabetic nephropathy and chronic kidney disease. Previous reports have demonstrated that aldosterone has a pathogenic role in renal injury via reactive oxygen species (ROS), which involves the regulation of autophagy. However, whether aldosterone can induce autophagy in renal tubular cells remains to be elucidated. In the present study, elevated autophagy was observed in rat renal tubular NRK-52E cells exposed to aldosterone, which was demonstrated by the increased number of autophagosomes, conversion of LC3-I to LC3-II and the expression of Beclin-1. The enhanced autophagy was accompanied by increased production of intracellular ROS, which was reversed by N-acetylcysteine, a specific inhibitor of ROS signaling. Furthermore, treatment with ginsenoside Rg1 reduced the aldosterone-induced autophagy and production of ROS, possibly through reducing the phosphorylation of AMPK and preserving mTOR activity. These findings demonstrated that aldosterone promoted ROS generation and increased autophagy in the NRK-52E cells. Ginsenoside Rg1 effectively relieved aldosterone-induced oxidative stress and abnormal autophagy, suggesting that Rg1 may be used as a potential therapeutic drug to inhibit the renal injury, which is induced by aldosterone.

Biodegradable and thermosensitive monomethoxy poly(ethylene glycol)-poly(lactic acid) hydrogel as a barrier for prevention of post-operative abdominal adhesion.

Post-operative peritoneal adhesions are serious consequences of abdominal or pelvic surgery and cause severe bowel obstruction, chronic pelvic pain and infertility. In this study, a novel nano-hydrogel system based on a monomethoxy poly(ethylene glycol)-poly(lactic acid) (MPEG-PLA) di-block copolymer was studied for its ability to prevent abdominal adhesion in rats. The MPEG-PLA hydrogel at a concentration of 40% (w/v) was injected and was able to adhere to defect sites at body temperature. The ability of the hydrogel to inhibit adhesion of post-operative tissues was evaluated by utilizing a rat model of abdominal sidewall-cecum abrasion. It was possible to heal wounded tissue through regeneration of neo-peritoneal tissues ten days after surgery. Our data showed that this hydrogel system is equally as effective as current commercialized anti-adhesive products.

[Management of chronic kidney disease guided by the theory of Traditional Chinese Medicine: an experimental study].

OBJECTIVE: To determine the impact of Traditional Chinese Medicine on patients with chronic kidney disease (CKD). METHODS: A total of 225 CKD patients in an outpatient department were recruited for this study, among whom 170 received regular Western and Chinese medicine treatments (control group) and 55 received treatments guided by the theory of Traditional Chinese Medicine (experimental group). The effectiveness of the treatments was determined through a pre-post comparison. RESULTS: Significant pre-intervention differences in age (P < 0.01), stage of glomerular filtration rate (GFR) (P = 0.007) and urine protein (P < 0.01) were found between the two groups of patients. But age, gender and proteinuria were not significant predictors on clinical outcomes of the patients in the multivariate regression models. The experimental group had a greater level of decrease in blood urea nitrogen (P < 0.01) and serum creatine (P < 0. 01) than the control group. No significant differences between the groups were found in changes of uric acid (P = 0.475), urine protein (P = 0.058), urine red cells (P = 0.577), and urine white cells (P = 0.01). A greater level of increase in estimated glomerular filtration rate was found in the experimental group compared with the control (P < 0.001). The multivariate linear regression analysis identified group (B = 0.395, P < 0.001) and stage of GFR (B = 0.165, P = 0.008) as significant predictors on the outcomes of treatment. CONCLUSION: The treatment of CKD patients guided by the theory of Traditional Chinese Medicine can improve renal function through influencing glomerular filtration rate. The effect is more prominent than the regular treatment regime.

[Investigate the effects of compound radix notoginseng on renal interstitial fibrosis and kidney-targeting treatment].

OBJECTIVE: Investigate the effects of compound Radix Notoginseng on renal interstitial fibrosis and kidney-targeting treatment. METHODS: 100 healthy Sprague-Dawley rats were randomly divided into 5 groups: Unilateral ureteral obstruction (UUO) group, sham-operation (SOR) group, Radix Notoginseng (RN) group, compound Radix Notoginseng (CRN) group and Losartan (ARB) group. After operation, RN, CRN and ARB groups were intragastric administrated with RN (3 mL/d), CRN (3 mL/d) and ARB [20 mg/(kg x d)] respectively. Each group randomly included 18 rats for statistical analysis. The histological changes of renal interstitial tissues were observed by HE, Masson and PAS staining. Total kidney collagen content was determined by measuring the amount of hydroxyproline. The mRNA of alpha-SMA, collagen I and fibronectin were reverse transcribed and quantified by real-time PCR. The expression of alpha-SMA protein was assessed by immunohistochemistry and Western blot analysis. RESULTS: In UUO model, the obstructed kidney showed typical features of renal tubulointerstitial fibrosis, such as severe tubular loss, dilation, atrophy, infiltration of inflammatory cells, interstitial matrix deposition (P < 0.05). Partial correlation assay showed that the expression of alpha-SMA was related to the renal tubular injury (r = 0.55; P < 0.05). Administration of RN, CRN and ARB improved tubulointerstitial damage and collagen matrix accumulation induced by UUO in different degree. The expression of the alpha-SMA at mRNA and protein levels were significantly increased in the UUO group (P < 0.05), which was also suppressed by treatment with RN, CRN and ARB in different degree. Moreover, more effective role in preventing fibrosis was observed in CRN group than when compared with that of RN group. CONCLUSION: RN and CRN can inhibit UUO-induced renal interstitial fibrosis in rats, and CRN treatment is more effective than RN in reducing interstitial fibrosis.

Ginsenoside-Rg1 protects podocytes from complement mediated injury.

AIM OF THE STUDY: Podocytes injury mediated by complement complex C5b-9 is the main feature of membranous nephropathy (MN). Little work has been done to prove that ginsenoside-Rg1 could inhibit this process. Our study aims to investigate the efficacy of ginsenoside-Rg1 in protecting the podocyte from complement mediated injury. MATERIALS AND METHODS: We chose sublethal C5b-9 induced podocyte injury as the model of MN in vitro. Ginsenoside-Rg1 was given as an intervention. Morphological changes were observed by electron microscope and fluorescence microscope. The production of reactive oxygen species (ROS) was detected by flow cytometry. The expression of the mitogen activated protein kinase (MAPK) including JNK, ERK and P38 was detected by western-blot technique. RESULTS: Ginsenoside-Rg1 could protect foot processes of podocytes, suppress the damage of F-actin, decrease the production of ROS, and inhibit the activation of P38 kinase pathway. CONCLUSION: These results suggest that ginsenoside-Rg1 could protect podocyte from sMAC-induced injury partly because of its antioxidant property and inhibit the activation of P38 kinase pathway.

Ginsenoside Rg1 modulation on thrombospondin-1 and vascular endothelial growth factor expression in early renal fibrogenesis in unilateral obstruction.

Renal interstitial fibrosis is the major histopathological change seen in a variety of renal disorders and is closely related to renal dysfunction. Progressive interstitial fibrosis accompanied by the loss of renal tubules and interstitial capillaries typifies all progressive renal disease. Thrombospondin-1 (TSP-1) is a major angiogenic inhibitor. It is demonstrated that TSP-1 levels were correlated with the loss of glomerular and peritubular capillaries and TSP-1 could promote renal scarring by effects on the endothelium. It has been reported that ginsenoside Rg1 inhibited renal interstitial fibrosis in rats via suppressing the expression of TSP-1. The present study was designed to examine whether ginsenoside Rg1 could modulate the integrity of the microvasculature and hence affect the progression of renal fibrosis in a rat unilateral ureteral obstruction (UUO) model. In UUO control kidneys, associated with interstitial fibrosis, lower peritubular capillary densities were prominent. These changes were all improved by ginsenoside Rg1 treatment. Interestingly, ginsenoside Rg1 decreased the expression of TSP-1 and enhanced vascular endothelial growth factor (VEGF) expression. The results show for the first time that ginsenoside Rg1 can evidently inhibit renal interstitial fibrosis in rats with UUO. The mechanism might be related to suppression of the expression of TSP-1 and to repair of the peritubular capillary.

[Effect of ginsenoside Rgl on the expression of TNF-alpha and MCP-1 in rats with diabetic nephropathy].

OBJECTIVE: To investigate the effects of Ginsenoside Rgl on proteinuria and the expression of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-alpha) in rats with diabetic nephropathy (DN). METHODS: The DN rat model was established by injection of streptozotocin (STZ, 65 mg/kg) in abdominal cavity. Forty Sprague-Dawley male rats were randomly divided into 4 groups: normal group, DN group, Ginsenoside Rgl treatment group and Irbesartan treatment group. The blood glucose was monitored routinely. Twenty-four hours urine protein and serum creatine were measured the day before the rats were killed when the eight weeks of treatments had been completed. The renal pathological and podocyte changes were evaluated. Immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) were performed to examine the protein expression levels of MCP-1 and TNF-alpha, respectively. The mRNA of TNF-alpha and MCP-1 were reverse transcribed and quantified by real-time PCR. RESULTS: The DN rats had increased volume of renal glomerulus, thickened basement membrane, and increased mesenterium mass, as well as some inflammatory cells in renal glomerulus. The number of potocyte decreased significantly in the DN group compared with the normal group (P<0.01). Compared with the DN group, the basement membrane became thinning and the number of podocyte increased in the two treatment groups (P<0.05). The rats in the DN group and the two treatment groups had significantly higher levels of twenty-four hour urine protein, serum creatine, serum glucose, serum MCP-1 and TNF-alpha than the normal rats (P<0.05). The rats in the treatment groups had lower levels of twenty-four hours urine protein and serum creatine than the rats in the DN group (P<0.05). But the serum glucose had little changes (P>0.05). There was no difference between the two treatment groups. Immunohistochemisty, ELISA and real-time PCR results indicated that the expression levels of MCP-1 and TNF-alpha in the rats in the DN group and the two treatment groups were significantly higher than those in the normal group (P<0.05). The rats in the treatment groups had lower levels of expression of MCP-1 and TNF-alpha than those in the DN group (P<0.05). The correlation analysis indicated that the levels of MCP-1 and TNF-alpha were positively related to twenty-four hours urine protein (r=0.7802, 0.6963), glomerular sclerosis index (r=0.8296, 0.7413) and thickness of podocyte membrane (r=0.7678, 0.6701, P<0.05). CONCLUSION: Ginsenoside Rgl reduces the expression of MCP-1 and TNF-alpha, repairs the pathological lesions of podocyte and nephron, and reduces the twenty-four hour urine protein rats with diabetic nephropathy.

Astragalus mongholicus ameliorates renal fibrosis by modulating HGF and TGF-beta in rats with unilateral ureteral obstruction.

Astragalus mongholicus (AM) derived from the dry root of Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is a widely used traditional Chinese medicine. The present study investigated the potential role of AM on renal fibrosis on a rat model of unilateral ureteral obstruction (UUO). We divided 48 Sprague-Dawley rats randomly into 4 groups: sham-operated group (Sham), untreated UUO group, AM-treated (10 g/(kg x d)) UUO group, and losartan-treated (20 mg/(kg x d)) UUO group as positive control. Haematoxylin & eosin (HE) and Masson staining were used to study the dynamic histological changes of the kidneys 7 and 14 d after operation. The expressions of fibronectin (FN), type I collagen (colI), hepatocyte growth factor (HGF), transforming growth factor-beta1 (TGF-beta1), and alpha-smooth muscle actin (alpha-SMA) were analyzed by real-time polymerase chain reaction (PCR), immunohistochemistry staining, and Western blot. Results show that, similar to losartan, AM alleviated the renal damage and decreased the deposition of FN and colI from UUO by reducing the expressions of TGF-beta1 and alpha-SMA (P<0.05), whereas HGF increased greatly with AM treatment (P<0.05). Our findings reveal that AM could retard the progression of renal fibrosis. The renoprotective effect of AM might be related to inhibition of myofibroblast activation, inducing of HGF and reducing of TGF-beta1 expression.

Ginsenoside Rb1, a panoxadiol saponin against oxidative damage and renal interstitial fibrosis in rats with unilateral ureteral obstruction.

OBJECTIVE: To investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction (UUO). METHODS: In total, 80 male rats were randomly divided into 4 groups, 20 in each group: the sham operated group (SOR), UUO group, UUO with ginsenoside Rb1 treatment group (treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group (as the positive control, treated with 20 mg/kg by gastrogavage per day). The rats were randomly sacrificed on day 3, 7 and 14 after surgery, respectively. The histopathologic changes of renal interstitial tissues were observed with Masson staining. The mRNA of transforming growth factor beta 1 (TGF-beta 1), collagen I and fibronectin were reversed transcribed and quantified by Real-time PCR. Enzyme-linked immunosorbent assay was used to quantitatively detect TGF-beta 1 and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. P47phox protein expression was assessed by immunohistochemistry and Western blot analysis. RESULTS: In the UUO model, the obstructed kidney showed typical features of progressive renal tubulointerstitial fibrosis, and the levels of TGF-beta1, collagen I and fibronectin increased (P<0.05). As compared with the UUO group, ginsennoside Rb1 significantly inhibited the interstitial fibrosis including tubular injury and collagen deposition, and decreased the levels of TGF-beta1 (P<0.05). Ginsenoside Rb1 also inhibited the heme oxygenase (HO-1) and 8-OHdG, two markers of oxidative stress (P<0.05). Moreover, ginsenoside Rb1 suppressed the expression of p47phox, a subunit of nicotinamide adeninedinucleotide phosphate (NADPH) oxidase (P<0.05). CONCLUSION: Ginsenoside Rb1 can obviously inhibit renal interstitial fibrosis in rats with UUO, its mechanism possibly via against the oxidative damage and suppressing TGF-beta1 expression.

[Meridian three-combined therapy for treatment of ordinary psoriasis: a multi-center randomized controlled study].

OBJECTIVE: To evaluate the effectiveness and safety of the meridian three-combined therapy for treatment of ordinary psoriasis. METHODS: A multi-central, randomized and positive drug controlled trial was adopted, and 233 cases were divided into an observation group of 116 cases and a control group of 117 cases. The observation group was treated with thread embedding at points, blood-letting puncture on the back of ear and auricular point pressing (i.e. meridian three-combined therapy). For thread embedding, 3-4 local points such ear points as Fei (CO14), Gan (CO12), Pizhixia (AT4), Shenmen (TF4) , cephalic and symmetric points of severe parts of the limb skin were selected according to the skin lesion position, and the treatment was given once each two weeks. For ear point tapping and pressing, 3-5 points were selected in each session. And the control group was treated with oral administration of Di yin Tablets, 5 tablets each time, twice each day. After treatment of 6 weeks, the clinical therapeutic effects, the score of skin lesion area, the scores for skin lesion severity and safety were compared in the two groups. RESULTS: The markedly effective rate was 57.8 % in the observation group and 51.3% in the control group with no significant difference between the two groups (P>0.05); after treatment the scores for both the skin lesion area and the skin lesion severity were significantly decreased in the two groups compared with those before treatment (P<0.01), and with a significant difference between the two groups (P>0.05). And the incidence rate of the adverse reaction in the observation group was significantly lower than that in the control group. CONCLUSION: The meridian three-combined therapy is effective and safe for treatment of ordinary psoriasis.

[Effect of Astragalus mongholicus on expression of hepatocyte growth factor in SD rats with unilateral ureteral obstruction].

OBJECTIVE: To study the effect of Astragalus mongholicus (AM) on the expression of hepatocyte growth factor (HGF) in SD rats with unilateral ureteral obstruction (UUO) and to elucidate the mechanisms underlying the renoprotective effects of AM. METHODS: Fifty four Sprague-Dawley rats were randomly divided into 3 groups: sham-operation group (SOR), UUO group (UUO) and UUO + AM group (AM). After administration of AM[10 g/ (kg x d)] for 3, 7 and 14 days,the histological changes of renal interstitial tissues were observed dynamically, and renal damage including tubular impairment and interstitial fibrosis were quantified on HE and Masson stained tissue sections. The protein expression of HGF and alpha-smooth muscle actin (alpha-SMA) was measured by immunohistochemistry. The mRNA expression of HGF and alpha-SMA were determined by real-time PCR. The expression of HGF and its receptor (C-met) were assessed by Western blot. RESULTS: Renal damage was exacerbated and the expression of alpha-SMA was significantly increased in UUO group compared with those of SOR group (P < 0.05) at each time point. HGF and C-met expression peaked at the 7th day after UUO and then decreased greatly. After AM intervention, tubular impairment and interstitial fibrosis were alleviated, up-regulations of alpha-SMA expressions was significantly suppressed, whileas the expression of HGF and C-met were significantly increased when compared with UUO group (P < 0. 05) at each time point. CONCLUSION: AM can ameliorate renal interstitial fibrosis induced by UUO in rats. The mechanisms of its antifibrotic effects may be related with the up-regulation of HGF and C-met expression, and the suppression of tubulo-epithelial mesenchymal transdifferentiation in renal intersitial progress.

[Effects of ginsenoside Rb1 on TGF-beta1 induced p47phox expression and extracellular matrix accumulation in rat renal tubular epethelial cells].

OBJECTIVE: To investigate the effects of Ginsenoside Rb1 (G-Rb1) on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-beta1 (TGF-beta1). METHODS: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, 10 ng/mL TGF-beta1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 (10 ng/mL, 20 ng/mL, 40 ng/mL) after TGF-beta1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI(diphenyleneiodonium, an inhibitor of NADPH oxidase) group. Intracellular reactive oxidative species (ROS) level was measured by flowcytometry. p47phox protein expression was assessed by immunohistochemistry and western blotting method. The expressions of collagen I (Col-I) and fibronectin(FN) gene were measured by real-time PCR analysis. The protein level of Col-I and FN were quantitatively detected by enzyme-linked immunosorbent assay. RESULTS: TGF-beta1 at 10 ng/mL significantly increased the intercellular ROS production and p47phox expression (P < 0.05). The levels of Col-I and FN were also significantly up-regulated with the stimulation of 10 ng/mL TGF-beta1 (P < 0.05). Compared to TGF-beta1-induced group, G-Rb1 and DPI depressed TGF-beta1-induced ROS production and p47phox overexpression. Meanhile, G-Rb1 and DPI decreased the levels of Col-I and FN. CONCLUSION: G-Rb1 could inhibit TGF-beta1 induced ROS production and decrease the levels of Col-I and FN in a dose-dependent manner. The mechanism might be partly related to the suppression of p47phox expression.

A case report of an effective treatment for diabetic foot ulcers with integration of traditional Chinese medicine and Western medicine.

Diabetes contributes 75-85% of the factors predisposing to foot amputations, usually in association with infection and gangrene. The treatment of foot ulcers is expensive, and the effectiveness of treatment varies. We report herein a case of a diabetic foot ulcer that was treated with integrated traditional Chinese and Western medicine, with desirable cost-effective results. Traditional Chinese medicine (TCM) therapeutic principles include improving the spleen, nourishing yin, regulating qi, and resolving dampness, as well as activating stagnant blood. Western medicine includes wound debridement, skin grafting, and use of insulin, antibiotics, and vasodilators. The patient was treated with a holistic multidisciplinary approach (i.e., a combination of TCM and Western medicine, surgical management, education for diabetic foot care, and psychological counseling). Without this approach, the patient might have ended up with foot amputation and/or sepsis.

Ginsenoside Rg1, a major active component isolated from Panax notoginseng, restrains tubular epithelial to myofibroblast transition in vitro.

The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-fibrosis properties. Epithelial-myofibroblast transition (EMT) plays an important role in renal tubulointerstitial fibrosis. The present study was designed to examine whether ginsenoside Rg1, a major active component isolated from Panax notoginseng, has an ability to block this phenotypic transition in rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-beta1 (TGF-beta1). The morphology of tubular epithelial-myofibroblast transition was observed through light microscope and transmission electron microscopy. alpha-SMA and E-cadherin are two markers of tubular epithelial-myofibroblast transition, their protein expressions were assessed by immunohistochemistry and western blot analysis. Gene expression of alpha-SMA as well as the two major extracellular matrix components collagen I and fibronectin was measured by real-time PCR analysis. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant. Our results revealed that ginsenoside Rg1 obviously blocked morphologic transformation in NRK-52E induced by TGF-beta1. Meanwhile, ginsenoside Rg1 inhibited the expression of alpha-SMA and the loss of E-cadherin, subsequently decreased the levels of collagen I and fibronectin in a dose-dependent manner. In addition, western blot analysis indicated that ginsenoside Rg1 inhibited the expression of P-ERK1/2 in NRK-52E induced by TGF-beta1. These results suggest that ginsenoside Rg1 can restrain the process of EMT maybe via suppressing the expression of P-ERK1/2 in vitro.

[Effect of Astragalus mongholicus on expression of connective tissue growth factor in kidney of SD rats with unilateral ureteral obstruction].

OBJECTIVE: To study the effect of Astrangalus mongholicus (AM) on the expression of Connective Tissue Growth Factor (CTGF) in SD rats with Unilateral Ureteral Obstruction (UUO) and elucidate the mechanism underlying the renorotective effects of AM. METHODS: 36 Sprague-Dawley rats were randomly divided into 3 groups: sham-operation group (Sham), UUO group (UUO) and UUO + AM group (AM). After administration of AM (10 g/kg x d) for 7 and 14 days, the dynamic histological changes of renal interstitial tissues were observed and renal damage including tubular impairment and interstitial fibrosis were quantified on HE and Masson stained tissue sections. The expressions of CTGF and alpha-smooth muscle actin (alpha-SMA) were measured by immunohistochemistry staining sections. The mRNA of CTGF and alpha-SMA were reversely transcribed and quantified to real-time PCR. The expression of CTGF protein was assessed by Western blot. RESULTS: Renal damage was exacerbated and the expressions of alpha-SMA and CTGF significantly increased in UUO group compared with those of Sham group (P < 0.05) at each time point. Tubular impairment and interstitial fibrosis were alleviated, and up-regulations of expressions of CTGF and alpha-SMA were significantly depressed by AM treatment (P < 0.05). CONCLUSIONS: AM can ameliorate renal interstitial fibrosis induced by UUO in rats. The mechanism of its antifibrotic effects may be related to the down-regulation of CTGF expression, following suppression of tubulo-epithelial mesenchymal transdifferentation in renal intersitial progress.

[Ginsenoside R(g1) inhibit transdifferentiation in rat renal tubular epethelial cells induced by TGF-beta1].

OBJECTIVE: To investigate the effects of ginsenoside R(g1) on the transdifferentiation of rat renal tubular epethelial cells induced by transforming growth factor-beta1, (TGF-beta1). METHOD: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, TGF-beta1-induced group and treated with ginsenoside R(g1) at different concentration (10, 20, 40 mg x L(-1)) group. The morphology of tubular epithelial-myofibroblast transdifferentiation induced by TGF-beta1 was observed through light microscope. alpha-SMA and E-cadherin protein expression were assessed by immunohistochemistry and western blot analyses. alpha-SMA, collagen I and and fibronectin gene expression were assessed by real-time quantitative chain reaction. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant. RESULT: 10 mg x L(-1) TGF-beta1 could induce the transdifferentiation of tubular epithelial myofibroblast, showing fibroblast-like in morphology, with significantly enhanced expression of alpha-SMA, depressed expression of E-cadherin and increased secretion of fibronectin and collagen I (P < 0.05). Compared to TGF-beta1-induced group, ginsenoside R(g1) partly abrogated the alpha-SMA expression and E-cadherin depression triggered by TGF-beta1 in tubular epithelial cells in a dose-dependent manner (P < 0.05). Meanhile, ginsenoside R(g1) blocked morphologic transformation of tubular epithelial cells and decreased levels of collagen I and fibronectin (P < 0.05). CONCLUSION: Ginsenoside R(g1) could inhibit TGF-beta1 induced the tubular epithelial-myofibroblast transdifferentiation and decreased levels of collagen I and fibronectin in NRK52E.