[Effects of zhenqing recipe and earthworm on the expressions of transforming growth factor-beta1 and plasminogen activator inhibitor 1 in renal tissues of type 2 diabetic rats].

OBJECTIVE: To study the effects of Zhenqing Recipe (ZQR) and earthworm on the expressions of transforming growth factor-beta1 (TGF-beta1) and plasminogen activator inhibitor-1 (PAI-1) in renal tissues of type 2 diabetes mellitus (T2DM) rats. METHODS: The T2DM model rats induced by high sucrose/fat feeding and low dose streptozotocin injecting were randomly divided into the model group, the ZQR group, and the earthworm group. A normal control group was also set up. ZQR extract was given at the daily dose of 24 g/kg body weight by gastrogavage to rats in the ZQR group. The earthworm extract was given at the daily dose of 2.6 g/kg body weight by gastrogavage to rats in the earthworm group. Equal volume of distilled water was given to rats in the normal control group and the model group by gastrogavage. The medication was once daily for 8 successive weeks. The fasting blood glucose (FBG), serum PAI-1, 24-h urinary albumin excretion (UAE), and endogenous creatinine clearance rate (CCr) were detected. The structural changes of renal tissues stained with Periodic Acid-Schiff were observed by light microscope. The expressions of TGF-beta1 and PAI-1 were analyzed by immunohistochemistry. And the mRNA and protein expressions of TGF-beta1 and PAI-1 in the renal cortex were detected by Real-time PCR and Western blot respectively. RESULTS: Compared with the normal control group, the FBG, serum PAI-1, UAE, the mRNA and protein expressions of TGF-beta1, and PAI-1 significantly increased in the model group, and CCr obviously decreased (P<0.01). Compared with the model group, FBG, serum PAI-1, UAE obviously decreased (P<0.01) and CCr obviously increased (P<0.05) in the ZQR group. FBG was not obviously lowered in the earthworm group. The serum PAI-1 and UAE both decreased obviously (P<0.05, P<0.01) and CCr increased (P<0.05) in the earthworm group. The effects of ZQR were superior to the earthworm. The renal pathological changes got improved in the ZQR group and the earthworm group. The mRNA and protein expressions of TGF-beta1 and PAI-1 significantly decreased in the ZQR group and the earthworm group, especially in the ZQR group. CONCLUSIONS: Early application of ZQR and earthworm in treatment of T2DM rats with nephropathy was effective in protecting the kidney. Its mechanism may possibly be correlated with inhibition of TGF-beta1 expression, resulting in down-regulation of PAI-1 expression. ZQR showed better effects than earthworm.

Co-Delivery of Paclitaxel and Interleukin-12 Regulating Tumor Microenvironment for Cancer Immunochemotherapy.

In the treatment of malignant tumors, the combination of chemotherapy that can directly kill tumor cells and immunotherapy that can activate the body's immune system and regulate tumor microenvironments is becoming one of the most promising cancer treatments. However, to co-deliver agents with different physicochemical properties for immunochemotherapy is still facing a challenge. Here, nanoparticles are developed for the co-delivery of the hydrophobic chemotherapeutic drug paclitaxel (PTX) and biomacromolecule interleukin-12 (IL-12) through the acid-sensitive material mPEG-Dlinkm -PDLLA and low-temperature expansion effect of Pluronic F127. The nanoparticles encrich in the tumor site, significantly inhibit the growth and metastasis of breast cancer cells 4T1, and prolong the overall survival of tumor-bearing mice. The underlying immune mechanism is further explored. The combination of PTX and IL-12 activates T lymphocytes and NK cells to release IFN-γ, selectively inhibits regulatory T cells and induces M1-type differentiation of tumor-related macrophages, thereby improving tumor immunosuppressive microenvironments. This study may provide an effective strategy for cancer immunochemotherapy through co-delivery of chemotherapeutic drug and immune cytokine by the facile thermo-sponge nanoparticles.

Role of salt inducible kinase 1 in high glucose-induced lipid accumulation in HepG2 cells and metformin intervention.

AIMS: To investigate the roles of salt inducible kinase (SIK1) in high glucose-induced triglyceride accumulation in human hepatoma HepG2 cells as well as in the molecular mechanism by which metformin, a drug to treat diabetes, suppresses high glucose-induced lipogenesis. MAIN METHODS: A cell model for high glucose-induced hepatic steatosis was prepared by exposing HepG2 cells to high glucose (25mmol) in the absence or presence of metformin (0.5mmol). Intracellular triglycerides were visualized by Oil Red O and measured using a triglyceride assay kit. Cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. SIK1 overexpression in HepG2 cells was achieved by transient transfection, and the mRNA and protein levels of SIK1 and lipogenic factors were measured using a reverse transcription-polymerase chain reaction and western blotting, respectively. KEY FINDINGS: Lipid accumulation in HepG2 cells was obvious after treatment with high glucose for 24h. In response to high glucose, SIK1 expression was negatively correlated with that of lipogenic factors and lipid accumulation in HepG2 cells. We observed that overexpression of SIK1, or treatment with metformin, suppressed lipogenesis, even in high glucose conditions. Furthermore, treatment with metformin upregulated SIK1 mRNA and protein levels, as well as the active form of SIK1. SIGNIFICANCE: SIK1 plays a vital role in high glucose-induced lipid accumulation, and metformin suppresses lipogenesis via the induction and activation of SIK1.

[Modulative effect of zhenqing recipe on expressions of matrix metalloproteinase-9 and tissue inhibitor of matrix metalloproteinase-1 in experimental type 2 diabetic rats].

OBJECTIVE: To explore the effect of Zhenqing Recipe (ZQR) on renal structure and expressions of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in experimental type 2 diabetic rats. METHODS: The rat models of type 2 diabetic were set up by intraperitoneally giving small-dose streptozotocin (STZ) after fed with high carbohydrate and high fat diets for one month. The model rats were randomly divided into the model group,the high and low dose ZQR-treated groups,and the enalapril-treated group; a normal control group was also established. The course of treatment continued 8 weeks. The expressions of MMP-9, TIMP-1, and fibronectin (FN) in renal tissues were detected by immunohistochemistry. The morphological changes of glomeruli and renal tubules were checked by microscopy. RESULTS: Compared with the normal control group, the expression of TIMP-1 and FN increased and MMP-9 decreased in the model group; the treated groups could decrease the expressions of TIMP-1 and FN, and increase the expression of MMP-9, especially the high-dose ZQR group had the best effect. The morphological changes of renal tubules and glomerulus in the treated groups were improved better as compared with the model group. CONCLUSION: The protective effect of ZQR on renal structure may be achieved by modulating the expressions of MMP-1 and TIMP-1.

[Determnination of betaine in Fufang Guilu granule by HPLC].

OBJECTIVE: To describe a HPLC method for assessing betaine in Fufang Guilu granule. METHOD: The content of betainephenaxcyl bromide in Fufang Guilu granule was determined by HPLC. The analytical column was a shim-pack CLC-ODS (6.0 mm x 150 mm) filling a 5 microm stationary phase; The mobile phase consisted of acetonitrile-water(35:65) with 0.1 mol x L(-1) NaClO4; The flow-rate was 1 mL x min (-1); The detector was set at 254 nm. RESULT: The calibration curve was linear over the range of 0.09-0.585 microg (r = 0.9997). The average recovery of the method was 98.4%, RSD 2.5% (n = 5). CONCLUSION: The results showed that this method was reliable and accurate, and can be used for quality control of Fufang Guilu granule.

Jueming Prescription reduces body weight by increasing the mRNA expressions of beta3-adrenergic receptor and uncoupling protein-2 in adipose tissue of diet-induced obese rats.

OBJECTIVE: To investigate the antiobesity effect of Jueming Prescription (JMP), a Chinese herbal medicine formula, and its influence on mRNA expressions of beta3 adrenergic receptor (beta3-AR) and uncoupling protein-2 (UCP-2) in adipose tissue of diet-induced obese rats. METHODS: Fifty male Sprague-Dawley rats were randomly divided into the normal control group (n =8) that was on a standard chow diet, and the obese model group (n =42) that was on a diet of high fat chow. Two weeks after the high fat diet, 29 obese rats in the obese model group were further randomly divided into 3 groups: the untreated obese model group (n =9), the metformin group (n =10, metformin 300 mg kg⁻¹ day)⁻¹, and the JMP group (n =10, JMP 4 g kg⁻¹ day⁻¹). After 8-week treatment, body weight, wet weight of visceral fat, and percentage of body fat (PBF) were measured. The levels of fasting blood glucose, serum lipids, and insulin were assessed, and insulin sensitivity index (ISI) was calculated. The adipose tissue section was stained with hematoxylin-Eosin, and the cellular diameter and quantity of adipocytes were evaluated by light microscopy. The mRNA expressions of beta3-AR and UCP-2 from the peri-renal fat tissue were determined by real-time reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Compared with the obese model group, treatment with JMP resulted in significantly lower body weight, wet weight of visceral fat, PBF, and diameter of adipocytes, and significantly higher level of high-density lipoprotein cholesterol, ISI (all P<0.01), JMP increased the mRNA expressions of beta3-AR and UCP-2 from perirenal fat tissue (P <0.05, P<0.01). CONCLUSIONS: JMP could reduce body weight and adipocyte size; and the effect was associated with the up-regulation of beta3-AR and UCP-2 expressions in the adipose tissue and improvement of insulin sensitivity.