Effects of novel Fufang Biejia Ruangan Tablets with sheep placenta as substitute for Hominis Placenta on CCl / 中草药·英文版

Objective: Fufang Biejia Ruangan Tablet (FBRT) is widely used for the treatment of liver fibrosis. However, Hominis Placenta (HP), as an important adjuvant of FBRT, has been restricted for medicinal using due to the limited availability, ethical controversy and safety issues. The present study aimed to investigate the therapeutic effects of novel FBRT (N-FBRT) with sheep placenta (SP) as substitute for HP on liver fibrosis and explore its possible mechanisms. Different dosages of SP in N-FBRT were also evaluated. Methods: Rats were subcutaneously injected with CCl

Mechanism of Fuzheng Qufeng Prescription in Suppressing Epithelial-mesenchymal Transition of Podocytes in Membranous Nephropathy Rats via TGF-β1/Smad Pathway / 中国实验方剂学杂志

Objective:To study the effect of Fuzheng Qufeng prescription （FZQP） on transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁）/Smad signaling pathway and epithelial-mesenchymal transition of podocyte in membranous nephropathy （MN） rats and to explore its molecular mechanism for podocyte protection. Method:The rats were randomly divided into normal control group （NC） and modeling group. Rats in modeling group induced by bovine serum albumin （C-BSA） were randomly divided into model group （MN）， losartan potassium group （LP， 0.05g·kg^-1）， and FZQP high dose （FZQPH， 41 g·kg^-1）， medium dose （FZQPM， 20.5 g·kg^-1）， and low dose （FZQPL， 10.25 g·kg^-1） groups. The administration lasted for 4 weeks. In week 0， 2， and 4 of administration， the levels of 24 hours urine protein （24 h-Upro） were tested. At the end of 4th week， the levels of blood urea nitrogen （BUN） and serum creatinine （SCr） were detected， and the rats in each group were sacrificed and the renal pathological morphology changes were observed by light microscope with hematoxylin-eosin （HE）， Masson and periodic acid-silver metheramine （PASM） staining. The deposition of immune complex， the thickening of glomerular basement membrane （GBM） and podocyte foot process were observed by transmission electron microscope （TEM）. The distribution and expression intensity of Desmin in renal tissues were detected by immunohistochemistry （IHC）. The mRNA and protein expression levels of TGF-<italic>β</italic>₁， Smad2/3， phospho（p）-Smad2/3， Smad7 and Desmin in renal tissues were respectively detected by Western blot （WB） and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with NC group， the levels of 24 h-Upro， BUN and SCr significantly increased in model group （<italic>P</italic><0.01）， with increased deposition of immune complex， significantly thickened GBM and fusion of foot processes， significantly increased Desmin mRNA and protein expression （<italic>P</italic><0.01） and increased TGF-<italic>β</italic>₁， Smad2， and Smad3 mRNA and protein expression （<italic>P</italic><0.05）， and decreased Smad7 mRNA and protein expression （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with model group， 24 h-Upro and BUN decreased in FZQP groups and LP group （<italic>P</italic><0.05）， levels of serum SCr in FZQPM group decreased （<italic>P</italic><0.05）， deposition of immune complex， thickening of GBM and fusion of foot process were all alleviated in FZQP groups and LP group. Distribution of Desmin along GBM decreased in FZQPH group， FZQPM group and LP group （<italic>P</italic><0.05）. Both mRNA and protein expression levels of TGF-<italic>β</italic>₁ and p-Smad2/Smad2 in FZQPM group decreased， while mRNA and protein expression levels of Smad7 increased （<italic>P</italic><0.05）. Both mRNA and protein expression levels of p-Smad3/Smad3 in FZQPH group decreased （<italic>P</italic><0.05）. Both mRNA and protein expression levels of Desmin in podocyte in FZQPH group， FZQPM group and LP group decreased （<italic>P</italic><0.05）. Conclusion:FZQP might realize podocyte protection effect in MN via suppressing EMT mediated by overactivated TGF-<italic>β</italic>₁/Smad signaling pathway.

Effect of Bushen Shugan Prescription on Oxidative Stress and TGF-β1/Smad Signaling Pathway in Hippocampus of Senile Depressed Mice / 中国实验方剂学杂志

Objective:To observe the changes in oxidative stress and transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁）/Smad signaling pathway in hippocampal tissue of senile depressed mice after chronic unpredictable mild stress and to explore the possible anti-depression mechanism of Bushen Shugan prescription. Method:Ninety five-month-old mice were randomly divided into six groups， namely the normal group， senile depression model group， high-， medium-， and low-dose Bushen Shugan prescription groups， and fluoxetine group， with 15 in each group. Mice in all groups， except for the normal group， were exposed to chronic unpredictable mild stress （CUMS） for inducing the senile depression. Since the first day of modeling， the mice in the high-， medium- and low-dose Bushen Shugan prescription groups were gavaged with 19.5， 9.75， 4.87 g·kg^-1 Bushen Shugan prescription， the ones in the fluoxetine group with 0.033 g·kg^-1fluoxetine， and those in the normal and senile depression model groups with an equal volume of normal saline for 21 successive days. The behavioral responses of mice in each group were evaluated in the open field test （OFT）， and the hippocampal tissues of mice were collected for testing the relevant indexes. The superoxide dismutase （SOD） content was determined by WST-1 method， malondialdehyde （MDA） content by TBA method， glutathione （GSH） content by micro enzyme-linked immunosorbent assay （ELISA）， and mRNA expression of TGF-<italic>β</italic>₁， Smad2， Smad3， and Smad7 by Real-time polymerase chain reaction （Real-time PCR）. Result:Compared with the normal group， the senile depression model group exhibited significantly lowered horizontal and vertical scores in OFT， decreased SOD and GSH contents in hippocampal tissues， elevated MDA （<italic>P</italic><0.05）， up-regulated TGF-<italic>β</italic>₁， Smad2， and Smad3 mRNA expression， and down-regulated Smad7 （<italic>P</italic><0.05）. Compared with the senile depression model group， Bushen Shugan prescription at the high， medium， and low doses and fluoxetine all increased SOD and GSH contents in mouse hippocampal tissues， decreased the MDA content （<italic>P</italic><0.05）， down-regulated the mRNA expression of TGF-<italic>β</italic>₁， Smad2， and Smad3 in hippocampal tissues， and up-regulated the Smad7 mRNA expression （<italic>P</italic><0.05）. The comparison with the high-dose Bushen Shugan prescription group showed that the SOD and GSH contents in hippocampal tissues of mice in the medium- and low-dose Bushen Shugan prescription groups declined significantly， while the MDA contents rose significantly （<italic>P</italic><0.05）. Besides， the mRNA expression levels of TGF-<italic>β</italic>₁， Smad2 and Smad3 in the hippocampal tissues of mice in the medium- and low-dose Bushen Shugan prescription groups were significantly up-regulated， and those of Smad7 were significantly down-regulated （<italic>P</italic><0.05）. Conclusion:Bushen Shugan prescription alleviates the depression symptoms in aged SAPM8 mice possibly by regulating the hippocampal oxidative stress and TGF-<italic>β</italic>₁/Smad signaling pathway.

Mechanism of Ethanol Extracts from Sophorae Tonkinensis Radix et Rhizoma on Hypertrophic Scars of Rabbit Ears Through TGF-β1/Smad Pathway / 中国实验方剂学杂志

Objective:To investigate that the effect of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma on the expression of transforming growth factor-β1 (TGF-β1)and Smad3 in the hypertrophic scars of rabbit ears and elucidate its mechanism to improve hypertrophic scars. Method:The model of hypertrophic ear scar model was established by damaging the inner skin of ears in New Zealand white rabbits.The 49 rabbits were randomly divided into control group, model group, low, medium and high-dose ethanol extracts groups from Sophorae Tonkinensis Radix et Rhizoma (0.4，1.0，2.0 g·kg-1), asiaticoside ointment group(5 mg·kg-1) and compound heparin sodium allantoin gel group(20 mg·kg-1), 7 rabbits per group. Except control group, the different drug about 0.5 mL had been applied the hypertrophic scar of rabbit ears once a day. After 42 days, the tissues of hypertrophic scar were obtained. Hematoxylin-eosin（HE）staining was used to observe the pathological changes of rabbit ear scar tissue and determine the scar hyperplasia index. The expression of TGF-β1 and Smad3 in scar tissue of rabbit ears were detected by immunohistochemistry, Western blot and reverse transcription PCR（RT-PCR）. Result:Compared with control group, the pathological changes of the ear scars in the model group showed obvious hyperplasia and higher hyperplasia index (P<0.01). Meanwhile, the expressions of TGF-β1 and Smad3 in scar tissue of rabbit ears were significantly increased (P<0.01). Compared with model group, the pathological structures of the ear scar tissue were significantly improved and the hyperplasia index of ear scar tissue was clearly reduced in medium and high-dose groups of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma(P<0.05，P<0.01). The protein and mRNA expression of TGF-β1 and Smad3 in scar tissue were also decreased in different group of ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma compared with the model group (P<0.05，P<0.01). Conclusions:Ethanol extracts from Sophorae Tonkinensis Radix et Rhizoma may play a curative role in inhibiting hypertrophic scars by reducing the expression of TGF-β1 and Smad3 in scar tissue and inhibiting the TGF-β1/Smads signal transduction pathway. These provides the experimental basis for the clinical application of Sophorae Tonkinensis Radix et Rhizoma in the treatment of hypertrophic scars.

Taxus chinensis ameliorates diabetic nephropathy through down-regulating TGF-β1/Smad pathway / 中国天然药物

Diabetic nephropathy (DN) is one of the common microvascular complications of diabetes mellitus. Renal fibrosis is closely related to the deterioration of renal function. The present study aimed to investigate protective effect of Taxus chinensis on high-fat diet/streptozotocin-induced DN in rats and explore the underlying mechanism of action. The rat DN model was established via feeding high fat diet for 4 weeks and subsequently injecting streptozotocin (30 mg·kg body weight) intraperitoneally. The rats with blood glucose levels higher than 16.8 mmol·L were selected for experiments. The DN rats were treated with Taxus chinensis orally (0.32, 0.64, and 1.28 g·kg) once a day for 8 weeks. Taxus chinensis significantly improved the renal damage, which was indicated by the decreases in 24-h urinary albumin excretion rate, blood serum creatinine, and blood urea nitrogen. Histopathological examination confirmed the protective effect of Taxus chinensis. The thickness of glomerular basement membrane was reduced, and proliferation of mesangial cells and podocytes cells and increase in mesangial matrix were attenuated. Further experiments showed that Taxus chinensis treatment down-regulated the expression of TGF-β1 and α-SMA, inhibited phosphorylation of Smad2 and Smad3. These results demonstrated that Taxus chinensis alleviated renal injuries in DN rats, which may be associated with suppressing TGF-β1/Smad signaling pathway.

Taxus chinensis ameliorates diabetic nephropathy through down-regulating TGF-β1/Smad pathway / 中国天然药物

Diabetic nephropathy (DN) is one of the common microvascular complications of diabetes mellitus. Renal fibrosis is closely related to the deterioration of renal function. The present study aimed to investigate protective effect of Taxus chinensis on high-fat diet/streptozotocin-induced DN in rats and explore the underlying mechanism of action. The rat DN model was established via feeding high fat diet for 4 weeks and subsequently injecting streptozotocin (30 mg·kg body weight) intraperitoneally. The rats with blood glucose levels higher than 16.8 mmol·L were selected for experiments. The DN rats were treated with Taxus chinensis orally (0.32, 0.64, and 1.28 g·kg) once a day for 8 weeks. Taxus chinensis significantly improved the renal damage, which was indicated by the decreases in 24-h urinary albumin excretion rate, blood serum creatinine, and blood urea nitrogen. Histopathological examination confirmed the protective effect of Taxus chinensis. The thickness of glomerular basement membrane was reduced, and proliferation of mesangial cells and podocytes cells and increase in mesangial matrix were attenuated. Further experiments showed that Taxus chinensis treatment down-regulated the expression of TGF-β1 and α-SMA, inhibited phosphorylation of Smad2 and Smad3. These results demonstrated that Taxus chinensis alleviated renal injuries in DN rats, which may be associated with suppressing TGF-β1/Smad signaling pathway.

Effect of Combined Intervention of Electroacupuncture and Astragaloside IV on Myocardial Hypertrophy and TGF-β 1/Smad Signaling in Rats with Myocardial Fibrosis / 针刺研究

OBJECTIVE: To observe the effect of combined intervention of electroacupuncture (EA) and astragaloside IV(ASIV) on cardiac hypertrophy and transforming growth factor β 1 (TGF-β 1)/Smad signaling in isoproterenol (ISO) induced cardiac hypertrophy rats, so as to investigate its underlying mechanisms in improving myocardial fibrosis. METHODS: A total of 50 SD rats were randomly divided into 5 groups: normal control, model (ISO), Propranolol (PRO)，ASIV and EA＋ASIV groups (n＝10 in each group). The myocardial fibrosis model was established by intraperitoneal injection (i.p.) of ISO (10 mg·kg－1·d－1), once daily for 30 days. Rats of the control group were given normal saline (i.p.), those of the PRO group given with PRO (40 mg·kg－1·d－1, gavage), and those of the ASIV and EA＋ASIV groups were treated by gavage of ASIV (40 mg·kg－1·d－1), once daily for 30 days. EA (20 Hz, 6 V) was applied to bilateral "Neiguan" (PC 6) for 10 min, once every day for 30 d. The heart mass index (HMI, whole heart weight/body weight) and left ventricular (LV) mass index (LVMI, weight of the LV/body weight) were calculated to assess the state of cardiac hypertrophy. The enzyme linked immunosorbent assay (ELISA) was used to determine the levels of procollagen I carboxy-terminal propeptide (PICP，a marker of extracellular matrix remodeling) and carboxyterminal telopeptide of type I collagen (ICTP, a metabolite of type I collagen) in serum, and Western blot was used to test protein contents of TGF- β 1, Smad 2 / 3, Smad 4, Smad 7 in the left ventricle tissue of the heart. RESULTS: After modeling, the HMI and LVMI, serum PICP and ICTP contents and the expression levels of myocardial TGF-β 1, Smad 2/3 and Smad 4 proteins were significantly increased in the model (ISO) group (P 0.05). CONCLUSIONS: EA stimulation of PC 6 combined with ASIV can relieve cardiac hypertrophy and myocardial fibrosis in rats, which may be associated with its effects in regulating myocardial TGF-β 1/Smad signaling pathway.

Regulatory effects of total triterpenoid of Prunella vulgarisL. on activities of ERK and TGF-β1/Smad signaling pathway in protecting hepatic fibrosis in rats / 中国药理学通报

Aim To investigate the protective effects of total triterpenoid from Prunella vulgaris L. ( TTP) on CCl4-induced hepatic fibrosis in rats and its mecha-nism. Methods Rat liver fibrosis was induced by 50% CCl4 twice a week for 12 weeks. From the 5th week, all the therapeutic groups were treated with the TTP(25, 50, 100 mg·kg-1 ) and the colchicine (0. 1 mg· kg-1 ) respectively once a day for 8 weeks. At the end of the twelfth week, the levels of ALT, AST, HA, PCⅢ, CⅣ, MDA, SOD, GSH-Px, Hyp were measured . HE and Masson staining were used to evalu-late the degree of hepatic fibrosis. The mRNA expres-sion ofα-SMA, procollagen I, Smad2, Smad3, Smad7 in liver was detected by RT-PCR, and the p-ERK pro-tein expression was evaluated by Western blot. Results Compared with the model group, TTP(25, 50, 100 mg·kg-1 ) not only reduced serum content of ALT, AST, HA, PCⅢ, CⅣand Hyp, MDA in liver tissue, improved the morphologic changes of hepatic fibrosis, but also increased SOD and GSH-Px activity. Moreo-ver, it decreased the α-SMA, procollagen I, Smad2, Smad3 mRNA expression and increased Smad7 mRNA expression in liver tissues obviously. Furthermore, TTP reduced the protein expression of p-ERK. Conclusions TTP can protect rats from CCl4-induced liver fibro-sis. The mechanism of this process may involve inhibi-ting the expression of p-ERK and interference with TGF-β1/Smad signal transduction pathway.

Interventional effects of fosinopril on TGF-β1/Smad signaling pathway in glomerular mesangial cells / 临床儿科杂志

Objective To observe the effects of fosinopril(FOS)on secretion of ColⅠ,expression of Smad2、Smad7 mRNA in TGF-β1-induced glomerulomesangial cells(GMC)in rat model. Methods Rat glomerular mesangial cells were cultured in vitro,passages 3 - 10 cells were used in the study after identification,and the cells were divided into 3 groups:control group(Ctrl group),TGF-β1 group,and fosinopril group. Expression of Col Ⅰ in cell culture supernatant was detected by the enzyme-linked immunosorbent assay(ELISA)at 6 h,24 h and 48 h. Changes of Smad2,Smad7 mRNA expression were evaluated by fluorescent quantitation PCR. Results Glomerular mesangial cells had Col Ⅰ protein expression. Secretion of Col Ⅰ was significantly higher in TGF-β1 group than those in Ctrl group at each time point(P ＜ 0.01),however the Col Ⅰ was significantly lower in fosinopril group at all time points than that in TGF-β1 groups(P ＜ 0.05). Glomerular mesangial cells also had Smad2,Smad7 mRNA expressions. The expressions of Smad2,Smad7 mRNA were significantly higher in TGF-β1 group than those in Ctrl group at each time point. Expression of Smad2 mRNA was significantly lower in fosinopril group than that in TGF-β1 group at all time points,while the difference in Smad7 mRNA expression between TGF-β1 group and fosinopril group showed no statistical significance(P ＞ 0.05). Conclusions Fosinopril could inhibit the secretion of Col Ⅰ and expression of Smad2 mRNA in glomerular mesangial cells induced by TGF-β1,suggesting that fosinopril might delay glomerular sclerosis through inhibiting the expression of Smad2 in TGF-β1/Smad signaling pathway.