Study on pathogenesis of mice model of coronary heart disease with phlegm-blood stasis syndrome based on PPARγ pathway / 中草药

Objective: To explore the pathogenesis of phlegm-blood stasis syndrome in mice model of coronary heart disease based on PPAR gamma pathway. Methods: Healthy SPF C57BL/6J mice were used in the control group and the sham operation group, and ApoE-/-mice were used in the blood stasis group, phlegm turbid group and phlegm-blood stasis group. The phlegm turbid group and the phlegm-blood stasis group were fed with high-fat diet for 12 weeks, and the other groups were fed with normal feed for 12 weeks. At the end of the 8th week, the left anterior descending coronary artery was ligated in the blood stasis group and the phlegm-blood stasis group. The sham operation group was not ligated. The levels of IL-6, ET, Ang II and PPARγ in serum were measured by enzyme linked immunosorbent assay, the levels of PPARγ, ABCA1 and CD36 protein in liver tissue were detected by Western blotting, and the levels of CD40, MMP-9 and NF-κB protein in aorta were detected by immunohistochemistry. Results: Compared with sham operation group, there was no significant change in serum IL-6, the content of serum ET in the group of phlegm and blood stasis was increased significantly (P < 0.01), the content of Ang II in blood stasis group was increased significantly (P < 0.05), the content of serum Ang II in phlegm turbid group and phlegm-blood stasis group was increased significantly (P < 0.01), and the content of PPARγ was decreased. In liver tissue, the expression levels of PPARγ and ABCA1 protein in blood stasis group, phlegm turbid group and phlegm-blood stasis group were decreased significantly (P < 0.01), the expression of CD36 protein was increased. CD40, MMP-9 and NF-κB levels in aorta tissue were increased significantly (P < 0.01). Conclusion: The phlegm-blood stasis syndrome of coronary heart disease can cause more serious atherosclerotic plaque in the course of its onset. Its mechanism may be through activating PPARγ pathway.

Effects of buddleoside on hypertensive vascular remodeling through Ang II/AT1 signaling pathway in aorta / 中草药

Objective: The aim of this paper was to observe the effects of buddleoside on hypertensive vascular remodeling through Ang II/AT1 signaling pathway in aorta. Methods: We used SHR model to examine the blood pressure, vertigo time, histomorphology, and collagen fiber distribution of the aorta, and evaluate whether buddleoside could ameliorate the hypertensive vascular remodeling in vivo. Meanwhile, abnormal proliferation and migration of VSMCs induced by Ang II in vitro was used to identify the mechanism. The anti-proliferation effect of buddleoside in VSMCs was observed using MTT assay and crystal violet assay. The anti-migration effect in VSMCs was observed using monolayer-wounding and boyden chamber transwell assay. Furthermore, the protein expression of Ang II, AT1, MMP-2, MMP-9, Src, p-Src, Syk, and p-Syk were examined. Results: The results showed that buddleoside could significantly decrease SBP, DBP, MBP, and vertigo time, and improve the thickened media aorta, hypertrophy and disordered arrangement of VSMCs, distribution of collagen fibers. Buddleoside could also inhibit the proliferation and migration of VSMCs, inhibit the ROS production, and reduce the protein expression of Ang II, AT1, MMP-2, MMP-9, Src, and p-Src. Conclusion: These data supported that buddleoside can ameliorate hypertensive vascular remodeling by inhibiting the proliferation and migration of VSMCs. Its mechanism is mediated by the regulation of Ang II/AT1 signaling pathway.

Protective Effect of Angiotensin (1-7) on Silicotic Fibrosis in Rats / 生物医学与环境科学（英文）

OBJECTIVE@#Silicosis, caused by inhalation of silica dust, is the most serious occupational disease in China and the aim of present study was to explore the protective effect of Ang (1-7) on silicotic fibrosis and myofibroblast differentiation induced by Ang II.@*METHODS@#HOPE-MED 8050 exposure control apparatus was used to establish the rat silicosis model. Pathological changes and collagen deposition of the lung tissue were examined by H.E. and VG staining, respectively. The localizations of ACE2 and α-smooth muscle actin (α-SMA) in the lung were detected by immunohistochemistry. Expression levels of collagen type I, α-SMA, ACE2, and Mas in the lung tissue and fibroblasts were examined by western blot. Levels of ACE2, Ang (1-7), and Ang II in serum were determined by ELISA. Co-localization of ACE2 and α-SMA in fibroblasts was detected by immunofluorescence.@*RESULTS@#Ang (1-7) induced pathological changes and enhanced collagen deposition in vivo. Ang (1-7) decreased the expressions of collagen type I and α-SMA and increased the expressions of ACE2 and Mas in the silicotic rat lung tissue and fibroblasts stimulated by Ang II. Ang (1-7) increased the levels of ACE2 and Ang (1-7) and decreased the level of Ang II in silicotic rat serum. A779 enhanced the protective effect of Ang (1-7) in fibroblasts stimulated by Ang II.@*CONCLUSION@#Ang (1-7) exerted protective effect on silicotic fibrosis and myofibroblast differentiation induced by Ang II by regulating ACE2-Ang (1-7)-Mas axis.

Farrerol inhibits AngII-induced VSMCs proliferation by reducing Cx43 expression / 中国药理学通报

Aim To study the role of Cx43 in inhibi-tion of AngII-induced vascular smooth muscle cells(VSMCs) proliferation by farrerol. Methods The primary VSMCs were isolated and cultured by direct adherent culture methods. VSMCs were identified by immunohistochemstry. The cells were divided into the following groups:control group,AngII group,AngII+Farrerol group. The cell viability was measured by CCK-8 cell vitality test. The proliferation of VSMCs was measured by the methods of Edu. The cell cycle of VSMCs was detected by flow cytometry. The mRNA levels of Cx43 were measured by Real-time PCR. The protein levels of Cx43 were measured by Western blot. Results 60 μmol·L-1farrerol could significantly de-crease the cell viability and EdU rate of VSMCs in-duced by AngII(P<0.05),which could also prevent the transformation of VSMCs from G0/G1phase to S phase. The results of real-time PCR and Western blot showed that,compared with the model group,Farrerol could significantly reduce the mRNA and protein ex-pression level of Cx43(P <0.01). After the interfer-ence of Cx43 by siRNA, the inhibition of proliferation by farrerol decreased significantly. Conclusion Far-rerol inhibits AngII-induced VSMCs proliferation signif-icantly, which might be associated with reducing the expression of Cx43.

Protective effect and mechanism of ligustrazine combined with astragaloside IV on angiogenesis of human umbilical vein endothelial cells / 中草药

Objective: To study the protective effect and mechanism of ligustrazine combined with astragaloside IV on hypoxia injury of human umbilical vein endothelial cells (HUVECs). Methods: The model of hypoxia injury was established, and the cells for pharmacodynamics study were divided into five groups: control group, model group, ligustrazine group (80 μg/mL), astragaloside IV group (40 μg/mL), and compatibility of astragaloside IV and ligustrazine group. The effects of ligustrazine, astragaloside IV, and their compatibility on cell proliferation in each group after hypoxia injury were detected by MTT assay. Immunohistochemical method was used to observe the expression of proteins VEGF and Ang-II in HUVECs with hypoxia injury, and Western blotting was used to observe the expression of proteins VEGF and Ang-II. RT-PCR was used to observe the mRNA expression of VEGF and Ang-II. Results: Compared with those in the model group, cell viability of ligustrazine group, astragaloside IV group, and compatibility group significantly increased, and the best group was ligustrazine (80 μg/mL)+astragaloside IV (40 μg/mL) group. Ligustrazine (80 μg/mL)+astragaloside IV (40 μg/mL) group could up-regulate the protein expression levels of VEGF and Ang-II and the levels of VEGF and Ang-II mRNA. Conclusion: Ligustrazine combined with astragaloside IV may be targeted by increasing angiogenesis factor, and the expression of VEGF and Ang-II plays the role of promoting angiogenesis.

Comparative effects of different extracts of Kansui Radix stir-baked with vinegar on function of expelling water retention with drastic purgative in cancerous ascites model rats / 中草药

Objective: To study the effects of different extracts of Kansui Radix stir-baked with vinegar (KRV) on the function of expelling water retention with drastic purgative in cancerous ascites model rats. Methods: The cancerous ascites model rats were respectively ig administered with KRV powder, ethanol extract, aqueous extract, and ehanol and aqueous extract of KRV (340 mg/kg) for 7 d. The amounts of urine and ascites, the levels of urinary sodium, potassium, chloride ion, and pH value, and the contents of PRA, Ang II, and ALD in serum were investigated. UPLC-QTOF MS technology was used to explore the components differences in various extracts of KRV. Results: Compared with the control group, the amount of urine in model group was significantly reduced (P<0.05), the ascites generated, and the urinary sodium, potassium, chloride ion, pH value, and the contents of PRA, Ang II, and ALD in serum were all significantly increased (P<0.01). Compared with the model groups, the treatment groups showed decreasing trend in ascites; The amounts of urine in positive groups, powder groups, ethanol and aqueous extract groups showed a significant increase (P<0.05); The level of urinary sodium of water extraction groups showed significant decrease (P<0.05); The levels of urinary sodium, potassium, chloride ion, pH value, and the contents of PRA, Ang II, and ALD in serum of positive groups, powder groups, ethanol extract groups, and ethanol and aqueous extract groups all showed a significant decrease (P<0.05, 0.01). Diterpenes were inspected in the alcohol extract and alcohol and aqueous extract, fewer in the aqueous extract. Conclusion: Powder groups and ethanol and aqueous extract groups of KRV have remarkable effect on expelling water retention with drastic purgative, and there is no significant difference between the two groups, which could provide the basis for clinical medication of KRV that is made into the pill and powder. Diterpenes in KRV may be the active components on the function of expelling water retention with drastic purgative.

Antagonism of toll-like receptor 2 attenuates the formation and progression of abdominal aortic aneurysm

Abdominal aortic aneurysm (AAA) is an inflammatory vascular disorder with high mortality. Accumulating evidence shows that toll-like receptor 2 (TLR2) plays a critical role in the regulation of wound-repairing process after tissue injury. We wondered if TLR2 signaling contributed to the pathogenesis of AAA and that targeting TLR2 would attenuate AAA development and progression. In this study, enhanced expression of TLR2 and its ligands were observed in human AAA tissue. Neutralization of TLR2 protected against AAA development and caused established AAA to regress in mouse models of AAA. In addition, TLR2-deficient mice also failed to develop AAA. The prophylactic and therapeutic effects of blocking TLR2 were accompanied by a significant resolution of inflammation and vascular remodeling, as indicated by the decreased expression or activity of MMP-2/9, α-SMA, inflammatory cytokines, and transcription factors NF-κB, AP-1 and STAT1/3 in AAA tissue. Mechanistically, blocking TLR2 decreased the expression and interaction of TLR2 and several endogenous ligands, which diminished chronic inflammation and vascular remodeling in the vascular tissue of AAA. Our studies indicate that the interactions between TLR2 and its endogenous ligands contribute to the pathogenesis of AAA and that targeting TLR2 offers great potential toward the development of therapeutic agents against AAA.

Antihypertensive effect and vascular regulation mechanism of rhynchophylline on spontaneously hypertensive rats / 中草药

Objective: To observe the effect of rhynchophylline (Rhy) on systolic blood pressure (SBP) with different time after administration in spontaneously hypertensive rats (SHR), and explore the protective effect on vascular endothelial cells of Rhy after long-term administration. Methods: Male SHR were randomly divided into model, positive control (Captopril 6.25 mg/kg), low-, mid-, and high-dose (1.25, 2.50, 5.00 mg/kg) Rhy groups. Other SD rats were included as the control group. Rats in the model and control groups were given the same volume of distilled water once daily for 21 d. Rat tail artery SBP was measured before administration and day 7, 14, and 21 during the administration. The levels of plasma Ang II, ADMA, AT1R, and serum NO, NOS were detected after the last administration underwent blood sampling. Results: Compared with the model group, Rhy reduced SBP significantly. Moreover, the plasma Ang II, ADMA, and AT1R levels were up-regulated, and the serum NO and NOS levels were decreased in the model group, which could be reversed by the treatment of Rhy (P<0.05, 0.01). Conclusion: Rhy could reduce the SBP of SHR significantly, decrease plasma Ang II, ADMA, and AT1R levels, and promote serum NO and NOS levels, which has the protection of vascular endothelial function.

The effects of Yishen-Pinggan decoction on the urine microalbumin and the expression of ACE2, AngⅡin the spotnaneously hype / 国际中医中药杂志

Objective To observe the effects of Yishen-Pinggan dcoction on the urine microalbumin and the expression of ACE2, AngⅡin the spontaneously hypertensive rat kidney(SHR);to observe the kidney level hepatic side of the protective effect of early renal in SHR hypertension. Methods The same old male SHR and normal rats(Wistar-Kyoto rats, WKY) for the study, at 10 weeks of age, SHR rats were randomly divided into a model group, a Chinese medicine group, and a lotensin group;while WKY rats were served as a normal control group(n=8). They were continuously treated for 12 weeks. Urine mAlb and β2-MG of rats were quantified at pre-experiment and after experiment, immuno-histochemical detection of renal ACE2 and AngⅡwere performed, the obtained results were statistically analyzed by computer image analysis. Result At the 12th week, compared with urinary mAlb(509.3±160.3)ng/ml,β2-MG(0.150±0.037)μg/ml, Ang kidneyⅡ(7 174.24±1 068.99)μm2,kidney ACE2(2 158.90±376.60)μm2 in the WKY group, the urinary mAlb(908.3 ±13.2)ng/ml, β2-MG(0.378±0.099)μg/ml, kidney AngⅡ(11 085.37±1 398.03)μm2 in the hypertension model group were significantly higher(P<0.01), and kidney ACE2(1 375.77±466.21)μm2 was decreased significantly(P<0.01); compared with hypertensive model group, rat urinary mAlb(574.9±197.8)ng/ml,β2-MG(0.198±0.040)μg/ml, kidney AngⅡ(8 462.91±781.23)μm2 in Chinese medicine group, rat urinary mAlb(644.4±147.5)ng/ml, β2-MG(0.206±0.044)μg/ml, kidney AngⅡ(8 696.88±679.70)μm2 in lotensin group were significantly decreased(P<0.01); while kidney ACE2(1 904.02 ± 454.08)μm2 in Chinese medicine group, kidney ACE2(1896.11±445.43)μm2 in lotensin group was significantly increased(P<0.01). Conclusion Yishen-Pinggan decoction can reduce the SHR urinary microalbumin excretion so as to reduce high blood pressure early kidney damage; its role may be associated with its increasing local renal ACE2 expression and reducing the kidney Ang Ⅱ expression.

Angiotensin II stimulates MCP-1 production in rat glomerular endothelial cells via NAD(P)H oxidase-dependent nuclear factor-kappa B signaling

Angiotensin II (Ang II) plays a crucial role in the pathogenesis of renal diseases. The objective of the present study was to investigate the possible inflammatory effect of Ang II on glomerular endothelial cells and the underlying mechanism. We isolated and characterized primary cultures of rat glomerular endothelial cells (GECs) and observed that Ang II induced the synthesis of monocyte chemoattractant protein-1 (MCP-1) in GECs as demonstrated by Western blot. Ang II stimulation, at concentrations ranging from 0.1 to 10 µm, of rat GECs induced a rapid increase in the generation of reactive oxygen species as indicated by laser fluoroscopy. The level of p47phox protein, an NAD(P)H oxidase subunit, was also increased by Ang II treatment. These effects of Ang II on GECs were all reduced by diphenyleneiodonium (1.0 µm), an NAD(P)H oxidase inhibitor. Ang II stimulation also promoted the activation of nuclear factor-kappa B (NF-κB). Telmisartan (1.0 µm), an AT1 receptor blocker, blocked all the effects of Ang II on rat GECs. These data suggest that the inhibition of NAD(P)H oxidase-dependent NF-κB signaling reduces the increase in MCP-1 production by GECs induced by Ang II. This may provide a mechanistic basis for the benefits of selective AT1 blockade in dealing with chronic renal disease.

Effect of Intravenous Low Intensity Laser Radiation Combined with TCM on Model Rabbit of Diabetic Stroke / 中国中医药信息杂志

Objective To observe the effect of intravenous low intensity laser radiation (ILLLI) combined with traditional Chinese medicine on TXB2, 6-Keto-PGF1? and Ang II of the rabbits of experimental diabetic stroke. Method 35 successfully modeled rabbits, after alloxian injection for diabetes and photochemical radiation for stroke, were randomized into four treatment group-control group (B), ILIB group (C), a group with compound treatment of ILIB and TCH (D), TCM treatment group (E), and 7 unmodeled rabbits were made as the normal group (A). TXB2, 6-Keto-PGF1? and Ang II level were observed and compared. Result Compared with group B, group C and E can significantly rectify the disorderly TXB2, 6-Keto-PGF1? and Ang II, Group D was better than C and E. Conclusion ILLLI combined with TCM can effectively rectify the TXB2, 6-Keto-PGF1? and Ang II level, reduce nervous injury, cure diabetes cerebral infarction.

Effect of Age on Proliferation of Rat Vascular Smooth Muscle Cells Stimulated with Fetal Bovine Serum, Angiotensin II and Phorbol 12-Myristate 13-Acetate / 대한해부학회지

The present study investigated effects of fetal bovine serum (FBS), angiotensin II (Ang II) and phorbol 12 -myristate 13 -acetate (PMA) on proliferation of vascular smooth muscle cells (VSMC) isolated from aorta of 5, 9, 12 and 17 week old Sprague -Dawley rats. With 10% FBS, proliferation rate of VSMCs was markedly greater in 17 week old group than in 5 week old group. Treatment of 0.1 ~10 micro M Ang II or 10 and 100 micro M PMA in 0.1% bovine serum albumin and 10% FBS media did not influence the cell proliferation. However, in 5% FBS media, Ang II and PMA enhanced the proliferation of the VSMC from all age groups except 5 week. To test the effects of Ang II in the FBS, captopril, an Ang converting enzyme inhibitor, and ibersartan, an Ang II receptor antagonist, were pretreated. Neither 10 micro M captopril nor ibersartan in 5% FBS influenced the VSMC proliferation. These results indicate that the VSMCs from older rats proliferate faster than those from 5 week -old rats, and 5% FBS -induced proliferation was enhanced by Ang II and PMA.