Scutellaria barbata D. Don inhibits growth and induces apoptosis by suppressing IL-6-inducible STAT3 pathway activation in human colorectal cancer cells.

One of the most critical cellular signal transduction pathways known to malfunction in colorectal cancer is the interleukin-6/signal transducer and activator of transcription 3 (IL-6/STAT3) pathway. Scutellaria barbata D. Don (SB) is well-known traditional medicine in China that targets STAT3 signaling, and it has long been used to treat various types of cancer; however, the precise mechanism of its antitumor activity remains largely unclear. In order to further elucidate this underlying mechanism, an ethanol extract of SB (EESB) in cancer treatment. The aim of the present study was to evaluate the effects of EESB on the IL-6-inducible STAT3 pathway. We tested the dose-response association between EESB, IL-6-induced proliferaion and apoptosis using an MTT assay, colony formation and flow cytometry analysis in vitro. In addition, caspase-9 and caspase-3 activation was determined using a colorimetric assay, the activity of IL-6-induced STAT3 pathway was evaluated using western blot analysis, and the expression levels of cyclin D1, cyclin-dependent kinase 4, Bcl2 and Bcl2-associated X were determined using reverse transcription-polymerase chain reaction and western blot analysis. In the present study it was found that EESB could significantly inhibit the IL-6-mediated increase in STAT3 phosphorylation levels and transcriptional activity in HT-29 human colon carcinoma cells, resulting in the suppression of cell proliferation and the induction of apoptosis. In addition, treatment with EESB markedly inhibited the IL-6-induced upregulation of cyclin D1 and B-cell lymphoma-2, two key target genes of the STAT3 pathway. These results suggest that treatment with EESB could effectively inhibit the proliferation and promote the apoptosis of human colon carcinoma cells via modulation of the IL-6/STAT3 signaling pathway and its target genes.

Prevalence and risk factors associated with prehypertension and hypertension in the Chinese She population.

BACKGROUND: Little is known about the prevalence and cardiovascular risk factors for prehypertension and hypertension in the She ethnic minority population of Fujian province in China. METHODS AND RESULTS: Between April 2009 and September 2009, 5,523 participants of She nationality aged between 20 and 80 years participated in this survey and 5,357 were eventually enrolled in analyses. The survey was carried out to assess blood pressure and cardiovascular risk factors. The prevalence of prehypertension and hypertension was 35.87 and 38.42%, respectively, in all participants. Only 26.63% of the subjects with hypertension were aware of their diagnosis. Multivariate logistic regression showed that age, gender, overweight/obesity, dyslipidemia and alcohol use were risk factors for prehypertension, and age, overweight/obesity, dyslipidemia, alcohol use, family history of hypertension and hyperuricemia were risk factors for hypertension. The clustering of 2 and ≥ 3 risk factors was in higher proportion for subjects with hypertension and prehypertension when compared with those with prehypertension and normotension, respectively. After adjusting for other confounding factors, multivariable logistic regression showed that the greater the number of clustering cardiovascular risk factors, the greater the odds ratios for prehypertension and hypertension are. CONCLUSION: Hypertension and prehypertension were common in the She population of Fujian province. Cardiovascular risk factors cluster during prehypertension and awareness of hypertension was minimal. Early lifestyle modifications could be advocated to prevent the transition from prehypertension to hypertension and cardiovascular disease.

Genetic variations in CYP17A1, CACNB2 and PLEKHA7 are associated with blood pressure and/or hypertension in She ethnic minority of China.

OBJECTIVES: Two large-scale genome-wide association studies (GWAs) have identified multiple variants associated with blood pressure (BP) or hypertension. The present study was to investigate whether some variations were associated with BP traits and hypertension or even prehypertension in adult She ethnic minority of China. METHODS: The population of the present study comprised 4460 (1979 males and 2481 females, respectively) unrelated she ethnic minority based on a cross-sectional study from Ningde City in Fujian province of China. There were 1692 hypertensives, 1600 prehypertensives and 1168 normotensive controls, respectively. We genotyped 7 variants in CYP17A1, PLEKHA7, CACNB2, ATP2B1, TBX3-TBX5, CSK-ULK3 and SH2B3 reported by the previous GWAs on Europeans. All analyses were performed in an additive genetic model. RESULTS: As the minor allele of rs653178 in/near SH2B3 was very rare with the frequency of 0.018, we excluded this single nucleotide polymorphism (SNP) in the further analyses. Of the other 6 loci, linear regression analyses revealed that rs11191548 in CYP17A1 and rs11014166 in CACNB2 were significantly associated with systolic BP (ß = -1.17, P = 0.002 and ß = -0.50, P = 0.006, respectively), while only SNP rs11191548 was significantly associated with diastolic BP (ß = -0.56, P=0.002) after adjusted by age, sex and BMI. Two variants in CACNB2 and PLEKHA7 were found to be significantly related to hypertension (odds ratios [OR] and (95% confidence interval [CI]): 0.79 (0.65-0.97) and 1.19 (1.01-1.41), respectively) in logistic regression analyses after adjusted by age, sex and BMI. In addition, we found that combined risk alleles of the 6 SNPs increased risk of hypertension in a stepwise fashion (P for trend < 0.001). However, none of the 6 SNPs was significantly associated with BMI or prehypertension status. While logistic analysis showed that subjects with cumulative risk alleles more than 9 had significantly higher risk for prehypertension (adjusted OR: 3.10, P < 0.001) compared with those with risk alleles less than 4. CONCLUSIONS: We replicated that variations in CYP17A1, CACNB2 and PLEKHA7 were related to BP traits and/or hypertension in She population. In addition, although we failed to observe single gene associated with prehypertension, we first found that conjoint effect of multiple risk alleles on BP might increase the risk of progressing to prehypertension.

Cardiovascular disease (CVD) risk, insulin resistance and ß-cell function in prehypertension population of China.

OBJECTIVE: To explore the cardiovascular disease (CVD) risk in prehypertensive subjects and evaluate whether high blood pressure (BP) is associated with insulin resistance (IR) and ß-cell dysfunction. METHODS: A total of 2949 people aged 20-94 years old were selected in Fujian province of China. We assessed CVD risk using Minnesota code-indicated major abnormal electrocardiography (MA-ECG) and presence of microalbuminuria in all population. IR/sensitivity and ß-cell function indices were derived from an oral glucose tolerance test. RESULTS: Prehypertensives with systolic/diastolic BP (SBP/DBP) 130-139/85-89 mm Hg had significant higher risk of MA-ECG and presence of microalbuminuria compared with normotensives (odds ratio [OR]: 1.483, 95% confidence interval [CI]: 1.016-2.165 and OR: 1.613, 95% CI: 1.142-2.277, respectively). In non-diabetic subjects, we found that prehypertensives and hypertensives had significant higher HOMA-IR and lower Matsuda's insulin sensitivity index compared with normotensives. There was a slightly decreased trend in ß-cell function assessed by disposition index (DIo) across the BP categories, when adjusted with age and BMI. The slight decline of DIo remained between hypertension and normotension, after additional adjustments were made, but the reduction of DIo lost statistic significance between prehypertension and normotension. CONCLUSIONS: Prehypertensives with SBP/DBP 130-139/85-89 mm Hg have higher CVD risk than normotensives. Prehypertension and hypertension are both in IR condition, however, what is more important is that early ß-cell dysfunction may exists in hypertension to some extent, while for prehypertension the compensation of ß-cell function may be appropriate.

The effect of NF-κB pathway on proliferation and apoptosis of human umbilical vein endothelial cells induced by intermittent high glucose.

Our previous study found that blocking nuclear factor (NF)-κB signaling could protect human umbilical vein endothelial cells (HUVECs) from apoptosis and proliferation inhibition due to high glucose (HG). Intermittent HG makes glucose toxicity more significant. In this study, we aimed to investigate the effect of NF-κB pathway on HUVECs induced by intermittent HG (a daily alternating 5.5 or 30.5 mmol/l glucose). A recombinant adenovirus containing a RNAi cassette targeting the NF-κB/p65 gene was produced, and its silencing effect on p65 gene was detected by Western blot analysis in HUVECs cultured with intermittent HG. The subsequent effect on proliferation of HUVECs in the indicated conditions was measured by the AlamarBlue assay. The Bcl-2 expression was also detected by Western blot. The results showed that the expression of p65 protein could be inhibited efficiently by the RNAi adenovirus. Intermittent HG also induced the translocation of NF-κB in HUVECs. Inhibition of NF-κB with the RNAi adenovirus could prevent the effects. At the 6th day after HUVECs were exposed to intermittent HG, the proliferation of HUVECs with Ad-1566 was significantly higher than that of HUVECs with Ad-DEST (P < 0.01). Knockdown of NF-κB/p65 up-regulated the Bcl-2 expression of HUVECs under intermittent HG conditions (P < 0.01). These findings concluded that the NF-κB/p65-targeting RNAi adenovirus is an important tool, which can efficiently inhibit the expression of p65 gene in HUVECs. Intermittent HG reduces HUVECs proliferation with a concomitant increase in apoptosis. Knockdown of NF-κB/p65 partly protected HUVECs from proliferation inhibition and may reduce apoptosis.

Construction of NF-κB-targeting RNAi adenovirus vector and the effect of NF-κB pathway on proliferation and apoptosis of vascular endothelial cells.

To construct a recombinant adenovirus vector expressing a RNAi for the Nuclear Factor kappa B (NF-κB)/p65 gene and use it to explore the role of the NF-κB pathway on the regulation of proliferation and apoptosis of vascular endothelial cells. A recombinant adenovirus containing a RNAi cassette targeting the p65 gene was constructed, and its silencing effect on p65 was detected by Western blot analysis in ECV304 cells. Expression of the p65 protein in ECV304 cells was efficiently down-regulated by the RNAi adenovirus for more than 6 days. ECV304 cells proliferation and apoptosis were measured using the MTT assay and flow cytometry, respectively. Blocking the NF-κB pathway with the RNAi adenovirus substantially decreased the proliferation of ECV304 cells, but only slightly affected cell apoptosis. We used a NF-κB/p65-targeting RNAi adenovirus to demonstrate the role of the NF-κB pathway in the regulation of ECV304 cell proliferation. This adenovirus may serve as an important tool to study the NF-κB pathway.

Associations between cardiovascular risk, insulin resistance, beta-cell function and thyroid dysfunction: a cross-sectional study in She ethnic minority group of Fujian Province in China.

OBJECTIVE: To investigate the associations between cardiovascular risk, insulin resistance (IR), ß-cell function and thyroid dysfunction in She ethnic minority group in China. METHODS: We enrolled 5080 participants of She ethnicity in this analysis eventually. We measured serum TSH and thyroid peroxidase antibody (TPOAb) concentrations, blood glucose and insulin levels in both fasting and 2-h postprandial states, serum lipid levels, blood pressure (BP), brachial-ankle pulse wave velocity (baPWV), electrophysiological parameters, including T(peak)-T(end) interval (T(p-e)), QT interval and height of the R wave in lead aVL (RaVL), and anthropometric parameters. RESULTS: The total prevalence of thyroid dysfunction in this population is 12.1%. Hyperthyroid subjects had shorter T(p-e) interval and QT interval in electrocardiogram (ECG), while hypothyroid subjects had shorter T(p-e) interval and longer QT interval in ECG than euthyroid subjects. Neither hyperthyroid nor hypothyroid subjects showed significant difference in BP, pulse pressure, and baPWV compared with euthyroid subjects. RaVL was slightly higher in hyperthyroid subjects, though the difference did not reach statistical significance (P=0.08). Subjects with TSH<0.3  mIU/l had higher blood glucose, insulin, homeostasis model assessment of insulin resistance (HOMA-IR), and ß-cell function (HOMA-ß), whereas subjects with TSH>10 mIU/l had lower insulin, HOMA-IR, and HOMA-ß than the reference group. There was a significant negative correlation, albeit weak, between TSH and HOMA-IR, HOMA-ß after adjustment for confounding factors. CONCLUSIONS: Hypothyroid subjects may carry higher cardiovascular risk than euthyroid subjects. Moreover, IR and ß-cell function are inversely correlated with TSH, which may be explained by the decreasing insulin-antagonistic effects of thyroid hormones along with increasing TSH.

Overweight, obesity, and their associations with insulin resistance and ß-cell function among Chinese: a cross-sectional study in China.

The aim of this study was to evaluate the associations of body mass index (BMI) with insulin resistance and ß-cell function in subjects with normal glucose tolerance. A cross-sectional study was carried out in Fujian province by multistratified sampling from July 2007 to May 2008. The sample consisted of 2931 subjects aged from 20 to 79 years. The questionnaires, physical examinations, and laboratory tests were obtained from all the participants. The homeostasis model assessment of insulin resistance (HOMA-IR) index was used to estimate insulin sensitivity, insulin secretion was assessed using the HOMA-ß index, and ß-cell function was quantified as the ratio of the incremental insulin to glucose responses over the first 30 minutes during the oral glucose tolerance test (ΔI30/ΔG30). Another measure was adjusted for insulin sensitivity as it modulates ß-cell function ([ΔI30/ΔG30]/HOMA-IR). Associations of BMI with morbidities were estimated using multiple logistic regression analysis. Relationships of BMI to insulin resistance and ß-cell function were assessed using multiple linear regression analysis and analysis of covariance. The age- and sex-adjusted prevalence of overweight and obesity was 23.04% (27.44% in men and 18.40% in women) and 2.65% (2.75% in men and 2.55% in women), respectively. After adjustment for covariables, BMI was independently associated with morbidity conditions; and there were increasing trend for odds ratios of morbidities across the BMI categories. There were independent differences for HOMA-IR, HOMA-ß, and ΔI30/ΔG30 between the normal-weight, overweight, and obese groups except for (ΔI30/ΔG30)/HOMA-IR. Body mass index was significantly and independently associated with HOMA-IR, HOMA-ß, and ΔI30/ΔG30 in the multiple linear regression analysis. Body mass index was an independent risk factor for hypertension, type 2 diabetes mellitus, dyslipidemia, metabolic syndrome, as well as the indexes of insulin resistance and ß-cell function. It is imperative that the whole society pay more attention to the identification and intervention of overweight and obesity to prevent obesity-related diseases at the very early stage.

High FFA-induced proliferation and apoptosis in human umbilical vein endothelial cell partly through Wnt/beta-catenin signal pathway.

Free fatty acids (FFA)-induced proliferation and apoptosis was studied in human umbilical vein endothelial cells (HUVECs). A recombinant adenovirus containing a RNAi cassette targeting the GSK-3beta gene was produced and its silencing effect on GSK-3beta gene was detected by Western blot analysis and immunohistochemistry assay in HUVECs. The effect of the RNAi on the protein level of beta-catenin was explored by transfecting the RNAi adenovirus to inhibit the expression of GSK-3beta protein. The subsequent effect on the Wnt/GSK-3beta/beta-catenin signal pathway and on proliferation and apoptosis of HUVECs cultured with FFAs, was analyzed by BrdU assay, Annexin V-FITC/PI Apoptosis Detection Kit, and 4',6-diamidino-2- phenylindole(DAPI) to explore the possible connection between the signaling pathway and FFA-induced proliferation and apoptosis. The Western blot results showed that the expression of GSK-3beta protein in HUVECs could be inhibited efficiently by the RNAi adenovirus, and that the protein level of beta-catenin was increased by RNAi adenovirus transfection. The results of the BrdU assay suggested that knockdown of GSK-3beta with the RNAi adenovirus may stimulate the proliferation of HUVECs. Apoptosis was observed in HUVECs exposed to FFAs (0.75 mmol/L) for 72 h, and this effect could be partly reversed when interfering with the RNAi adenovirus. It may be concluded that the RNAi adenovirus specific to GSK-3beta may partly protect HUVECs from apoptosis induced by FFAs, probably through the up-regulation of the Wnt/beta-catenin signal pathway.

Regulation of GSK-3 beta in the proliferation and apoptosis of human thyrocytes investigated using a GSK-3 beta-targeting RNAi adenovirus expression vector: involvement the Wnt/beta-catenin pathway.

Disorders in the proliferation and apoptosis of thyrocytes may induce goitre, adenoma and carcinoma in the thyroid. The Wnt/beta-catenin pathway has been demonstrated to be involved in the regulation of cell proliferation, differentiation and apoptosis in various cell lines. The regulatory mechanism on the proliferation and differentiation of thyrocytes is not well characterized. In the present study, a GSK-3beta-targeting RNA interference (RNAi) adenovirus vector was constructed and delivered to primary human thyrocytes. Results showed that the expression of beta-catenin protein in primary human thyrocytes was increased after GSK-3beta-targeting RNAi adenovirus infection, the proliferation of primary human thyrocytes was significantly stimulated using Bromodeoxyuridine (BrdU) assay, while cell apoptosis was slightly affected which was observed through flow cytometry. It is concluded that the Wnt/beta-catenin pathway plays a significant role in the regulation of the proliferation of primary human thyrocytes.