Qian Yang Yu Yin Granule protects against hypertension-induced renal injury by epigenetic mechanism linked to Nicotinamide N-Methyltransferase (NNMT) expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Qian Yang Yu Yin Granule (QYYY) is a Chinese herbal formulation. It is used to treat hypertensive nephropathy for decades in China, but it is unknown that the exact mechanism of QYYY on hypertensive nephropathy. AIMS OF STUDY: The present study was to elucidate its epigenetic mechanism of QYYY on hypertensive nephropathy. MATERIALS AND METHODS: In the current study, HEK293T cells' proliferation induced by Ang II was chosen to observe epigenetic mechanisms of QYYY on renal damage. The cell proliferation was examined by MTT assays and ethynyldeoxyuridine analysis. Cell cycle analysis was performed. After treatment with QYYY, expression of Nicotinamide N-methyltransferase (NNMT), sirtuin1(SIRT1), S-adenosylhomocysteine(SAH), histone H3K4 methylation, and cortactin acetylation(acetyl-cortactin,ac-cortactin) were further investigated by western-blotting and real time PCR. DNA methylation was detected by ELISA. The study also observed the changes of SIRT1, SAH, H3K4 methylation, acetyl-cortactin when NNMT over-expressed by lentivirus transfection. Angiotensin II(Ang II) induced renal damage in spontaneously hypertensive rats(SHR). After eight weeks treatment of QYYY, blood pressure, serum and urine creatinine, and urinary microalbumin(mAlb) were assessed. The concentration of N1 -methylnicotinamide were detected by liquid chromatography with tandem mass spectrometry. The protein of NNMT, ac-cortactin, H3K3me3 were also assessed in vivo. RESULTS: QYYY inhibited HEK293T cells' proliferation, down-regulated the expression of NNMT, SAH, acetyl-cortactin and DNA methylation, up-regulated the expression of SIRT1, histone H3K4 trimethylation(H3K4me3). Over-expression of NNMT increased the expression of SAH and acetyl-cortactin, and reduced the expression of SIRT1 and H3K4me3. The study also demonstrated that QYYY promoted urinary creatinine excretion and reduced serum creatinine and urinary mAlb in SHR. QYYY decreased the concentration of N1 -methylnicotinamide in Ang II group. QYYY decreased the protein of NNMT, ac-cortactin and increased H3K4me3 in vivo. CONCLUSION: The results showed that QYYY alleviated renal impairment of SHR and inhibited HEK293T cells' proliferation induced by Ang II through the pathway of epigenetic mechanism linked to Nicotinamide N-Methyltransferase (NNMT) expression, including histone methylation, DNA methylation and acetyl-cortactin. This study unveiled a novel molecular mechanism by which QYYY controlled the progression of hypertensive nephropathy.

YQWY Decoction Improves Myocardial Remodeling via Activating the IL-10/Stat3 Signaling Pathway.

Heart failure (HF) has been known as a global health problem, and cardiac remodeling plays an essential role in the development of HF. We hypothesized that YQWY decoction might exert a cardioprotective effect against myocardium inflammation, fibrosis, and apoptosis via activating the interleukin-10 (IL-10)/Stat3 signaling pathway. To test this hypothesis, the HF model in rats was established by pressure overload through the minimally invasive transverse aortic constriction (MTAC). Echocardiography was performed to assess the left ventricular function of rats. Myocardial fibrosis in rats was observed by Masson and Picrosirius red staining, and the degree of myocardial apoptosis was detected via TUNEL staining. In addition, expression levels of IL-10, tumor necrosis factor-α (TNF-α), Stat3 (P-Stat3), P65 (P-P65), CD68, collagen I, TGF-ß, CTGF, Bax, Bcl-2, cleaved caspase-3, and PARP in rat serum and myocardium samples were examined by ELISA, western blot, and immunohistochemistry, respectively. YQWY decoction treatment significantly improved left ventricular function in HF rats, especially in those of the high-dose group (LVEF%: 51.29 ± 5.876 vs. 66.02 ± 1.264, P < 0.01;, LVFS%: 27.75 ± 3.757 vs. 37.76 ± 1.137, P < 0.01). Furthermore, YQWY decoction markedly inhibited MTAC-induced myocardial fibrosis as evidenced by downregulated collagen I, TGF-ß, and CTGF in myocardium and alleviated apoptosis (downregulated caspase-3 and PARP and increased Bcl-2/Bax ratio in cardiomyocytes). In addition, YQWY decoction decreased the level of the proinflammatory cytokine TNF-α in both circulating blood and myocardium and attenuated infiltration of inflammatory cells in heart tissue from HF rats. Most importantly, YQWY decoction suppressed MTAC-induced NF-κB activation and phosphorylated Stat3 by upregulating IL-10 in rat heart tissues. Our study showed that YQWY decoction could attenuate MTAC-induced myocardial inflammation, fibrosis, apoptosis, and reverse the impairment of cardiac function in rats by activating the IL-10/Stat3 signaling pathway and improving myocardium remodeling. Our findings suggested a therapeutic potential of YQWY decoction in HF.

YQWY decoction reverses cardiac hypertrophy induced by TAC through inhibiting GATA4 phosphorylation and MAPKs.

To investigate the effect of Yiqi Wenyang (YQWY) decoction on reversing cardiac hypertrophy induced by the transverse aortic constriction (TAC). Wistar rats aged 7-8 weeks were subjected to TAC surgery and then randomly divided into 4 groups (n = 5/group): Sham group, TAC group, low-dose group and high dose group. After 16-week intragastric administration of YQWY decoction, the effect of YQWY decoction on alleviating cardiomyocyte hypertrophy was examined by transthoracic echocardiography (TTE), hematoxylin/eosin (HE), wheat germ agglutinin (WGA) staining, enzyme linked immunosorbent assay (ELISA), Western blot (WB), immunohistochemistry (IHC) and immunofluorescence (IF), respectively. The results showed significant differences in left ventricle volume-diastole/systole (LV Vol d/s), N-terminal pro-B-type brain natriuretic peptide (NT-proBNP) (P < 0.01), Ejection Fraction (EF), LV mass and fractional shortening (FS) (P < 0.05) between YQWY-treated group and TAC group. HE and WGA staining showed that treatment with YQWY decoction dramatically prevented TAC-induced cardiomycyte hypertrophy. Moreover, the results of WB, IHC and IF indicated that administration of YQWY could suppress the expressions of cardiac hypertrophic markers, which included the atrial natriuretic peptide (ANP), BNP and myosin heavy chain 7 (MYH7) (P < 0.05) and inhibit phosphorylation of GATA binding protein 4 (P-GATA4) (P < 0.05), phosphorylation of extracellular signal-regulated kinase (P-ERK) (P < 0.05), phosphorylation of P38 mitogen activated protein kinase (P-P38) (P < 0.05) and phosphorylation of c-Jun N-terminal kinase (P-JNK) (P < 0.05). Thus, we concluded that YQWY decoction suppressed cardiomyocyte hypertrophy and reversed the impaired heart function, and the curative effects of YQWY decoction were associated with the decreased phosphorylation of GATA4 and mitogen activated protein kinases (MAPKs), as well as the reduced expression of the downstream targets of GATA4, including ANP, BNP, and MYH7.

Panax quinquefolius saponin inhibits endoplasmic reticulum stress-mediated apoptosis and neurite injury and improves functional recovery in a rat spinal cord injury model.

The treatment goal in spinal cord injury (SCI) is to repair neurites and suppress cell apoptosis. Panax quinquefolius saponin (PQS) is the major active ingredient of American ginseng and has been demonstrated to have anti-inflammatory and anti-apoptotic roles in various diseases. However, the potential effect of PQS on the pathological process of acute SCI remains unknown. This work tested the effects of PQS on acute SCI and clarified its potential mechanisms. PQS treatment ameliorated the damage to spinal tissue and improved the functional recovery after SCI. PQS treatment inhibited endoplasmic reticulum (ER) stress and the associated apoptosis after acute SCI. PQS further abolished the triglyceride (TG)-induced ER stress and associated apoptosis in neuronal cultures. PQS appears to inhibit the ER-stress-induced neurite injury in PC12 cells. Our results suggest that PQS is a novel therapeutic agent for acute central nervous system injury.

[Research progress of traditional Chinese medicine intervention vascular cognitive impairment].

Vascular cognitive impairment (VCI) is a kind of syndrome from mild cognitive impairment to dementia, which is caused by different vascular factors. It can be prevented and delayed the progress of VCI and even reversed cognitive impairment before it progresses to vascular dementia by early diagnosis and intervention. Many experimental and clinical studies have confirmed that traditional Chinese medicine (TCM) monomer, effective fraction, compound preparation,etc can improve vascular cognitive function. Our paper summarizes the research progress in the concept, pathogenesis, cellular and molecular mechanisms, and TCM treatment of VCI.

[Effects of ApoE gene polymorphism on anti-inflammatory action of xuezhikang capsule].

OBJECTIVE: To study the effects of ApoE gene polymorphism on anti-inflammatory action of Xuezhikang Capsule. METHODS: One hundred and two patients with hyperlipidemia (as the treated group) and one hundred healthy volunteers (as the control group) were enrolled in the case-control study. Total DNA of the peripheral blood was extracted and ApoE genotypes were determined by PCR sequence analysis. The serum levels of tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and high sensitivity C reactive protein (hs-CRP)were measured in all subjects. The changes of TNF-alpha, IL-6, and hs-CRP were detected before and after 6-week Xuezhikang Capsule treatment, thus analyzing the correlation between ApoE gene polymorphism and changes of each inflammatory factor. RESULTS: The frequency of E3/3 genotype was 86% (86/100 cases)in the control group, significantly higher than that of the treated group (62.7%, 64/102 cases). The frequency of E3/4 genotype was 6% (6/100 cases) in the control group, significantly lower than that of the treated group (21.6%, 22/102 cases; both P < 0.05). Compared with the control group, the serum levels of TNF-alpha, IL-6, and hs-CPR were higher in the treated group before treatment (P < 0.05). In hyperlipidemia patients with E3/4 + E4/4 genotype, the serum level of TNF-alpha was higher than that of E3/3 genotype (P < 0.05); the serum level of IL-6 was higher than that of E2/E2 + E2/E3 genotype (P < 0.05); the serum level of hs-CRP was higher than that of E2/E2 + E2/E3 and E3/E3 genotype (P < 0.05). But there was no statistical difference in the serum levels of TNF-alpha, IL-6, or hs-CPR between E3/3 and E2/E2 + E2/E3 genotype. After 6-week intervention of Xuezhikang Capsule, the serum levels of TNF-alpha, IL-6, and hs-CRP were lower in the treated group (P < 0.05), but the serum levels of TNF-alpha and IL-6 were still higher than those of the control group (P < 0.05). But there was no statistical difference in the decrement of TNF-alpha, IL-6, or hsCRP among E2/E2 + E2/E3, E3/E3, or E3/4 + E4/4 genotypes (P > 0.05). CONCLUSIONS: The distribution of ApoE gene polymorphism is different between the hyperlipidemia patients and the healthy people. Chronic inflammatory reactions exist in hyperlipidemia patients, especially in those with e4 allele. Xuezhikang Capsule showed anti-inflammatory effects, but ApoE gene polymorphism did not affect its effects.

[Effect of Jiangzhi Kangyanghua mixture on high-sensitivity C-reactive protein and vascular endothelial functions of hypertension patients].

To observe the effect of Jiangzhikangyanghua Mixture on high-sensitivity CRP (hs-CRP) and vascular endothelial functions of essential hypertension (EH) patients. In this study, 72 cases of out-patients with EH were selected from department of cardiology of Wujin hospital of traditional Chinese Medicine, and randomly divided into the control group (n= 36, amlodipine 5 mg qd + valsartan 80 mg qd) and the test group (n =36 amlodipine 5 mg qd + valsartan 80 mg qd + Jiangzhikangyanghua mixture 20 mL tid). The contents of hs-CRP, ET-1 and NO were measured before and after treatment for two months. The result showed that the contents of hs-CRP, ET-1 in both groups reduced (P <0. 05) , while the test group show a more significant reduction than the control group (P <0. 05). After the treatment, the content of NO raised in both group, while the test group show a more significant increase than that of the control group (P <0. 05). This study indicated that Jiangzhi Kangyanghua mixture could reduce the contents of hs-CRP and ET-1 and raise NO of EH patients.

[Distribution of Apo E gene sequencing in patients with coronary heart disease of phlegm-stasis syndrome].

OBJECTIVE: To study the correlation between Apo E gene polymorphism and patients with coronary heart disease (CHD) of phlegm-stasis syndrome (PSS). METHODS: 78 CHD patients were assigned to PSS (49 cases) and non-phlegm-stasis syndrome (NPSS). Polymorphisms of Apo E gene in 78 CHD patients and 100 healthy subjects were detected by complete DNA sequencing. RESULTS: Five gene types as E3/3, E4/4, E2/ 3, E2/4, and E3/4 were detected in the two groups. The frequencies of genotype E3/3 and epsilon 3 allele were significantly lower in CHD patients than in the healthy subjects (P<0.01). But the frequencies of genotype E3/4 and epsilon 4 allele were significantly higher in CHD patients than in the healthy subjects (P<0.01). In CHD patients, the frequencies of genotype E2/4 + E3/4 + E4/4 and epsilon 4 allele were higher in PSS than in NPSS. CONCLUSIONS: Apo E epsilon 4 allele was a susceptible allele to CHD, which was closely correlated to CHD PSS. It was inferred that it might be one of main susceptible alleles for CHD PSS.

[Influence of apolipoprotein E gene polymorphism on the lipid regulation by xuezhikang capsule].

OBJECTIVE: To explore the influence of apolipoprotein E (ApoE) gene polymorphism on the lipid metabolism regulatory effect of Xuezhikang Capsule (XZKC). METHODS: ApoE polymorphism of 74 patients with hyperlipidemia was detected by gene sequencing method, and their plasma levels of total cholesterol (TC), triglyceride (TG), high- and low-density lipoprotein cholesterol (HDL-C and LDL-C) were determined before and after they received a 6-week treatment of XZKC, for analyzing the relationship between ApoE gene polymorphism and the changes of various blood lipids associated indices. RESULTS: The effect of XZKC on reducing TG in the epsilon2 allele (E2/E2 and E2/E3 genotypes) was higher than that in the E3/E3 genotypes and epsilon4 allele (E3/E4 and E4/E4 genotypes), while on increasing HDL-C, it showed more effect in the epsilon4 allele (E3/E4 and E4/E4 genotypes) than that in the epsilon2 allele (E2/E2 and E2/E3 genotypes) and E3/E3 genotypes. CONCLUSION: Patients' ApoE gene polymorphism could influence the lipid regulatory effect of XZKC, embodying mainly by raising HDL-C and reducing TG in patients with different ApoE genotypes to different extents.

[Study on the relationship between polymorphism of ApoE gene and TCM syndrome type of primary hyperlipemia].

OBJECTIVE: To study the relationship between the polymorphism of ApoE gene and TCM syndrome type of primary hyperlipemia. METHODS: ApoE genotype of 102 patients with hyperlipemia was detected by gene PCR sequencing. RESULTS: A total of five genotypes were detectable, they were E2/2, E3/3, E4/4, E2/3 and E3/4. The frequency of E3/4 + E4/4 and epsilon4 allelotype detected in the patients of Gan-Shen Yin deficiency syndrome type were significantly higher than those in patients of Pi-Shen Yang-deficiency type or of phlegm stagnation type (P < 0.05, P < 0.01), and which in patients of Qi-stagnation caused blood stasis type were significantly higher than those in patients of phlegm stagnation type ( P < 0.05). CONCLUSION: Polymorphism of ApoE gene is related in a certain degree to TCM syndrome type of primary hyperlipemia.