Ganzhirong Granule Inhibits Hepatic Gluconeogenesis through the SIRT3-MPC1-PC/PDH Axis in Type 2 Diabetes

Summary

This study investigated whether Ganzhirong Granule ameliorates hyperglycemia in type 2 diabetes by inhibiting hepatic gluconeogenesis through the SIRT3-MPC1-PC/PDH axis.

Abstract

Type 2 diabetes mellitus (T2DM) is a global metabolic disorder characterized by hepatic insulin resistance and excessive gluconeogenesis. Ganzhirong Granule (GZRG), a traditional Chinese medicine formula, has shown potential in managing T2DM, but its underlying mechanisms, particularly concerning hepatic gluconeogenesis, remain unclear. This study investigated whether GZRG ameliorates hyperglycemia by modulating the SIRT3-mediated signaling pathway. The anti-diabetic effects of GZRG were evaluated in high-fat diet (HFD)-induced T2DM mice and free fatty acid (FFA)-induced insulin-resistant HepG2 cells. Metabolic parameters, glucose and pyruvate tolerance, insulin sensitivity, and lipid profiles were assessed. Molecular mechanisms were explored through the overexpression and knockdown of SIRT3, examining the expression of key proteins (SIRT3, MPC1, PC, PDH-E2, PCK1, and G6Pase) in the gluconeogenic pathway. GZRG treatment significantly ameliorated hyperglycemia, enhanced insulin sensitivity, and improved lipid metabolism in both T2DM mice and insulin-resistant (IR) HepG2 cells. It attenuated hepatic steatosis and suppressed gluconeogenesis. Mechanistically, GZRG downregulated SIRT3 expression, which led to concomitant reductions in MPC1 and PC levels and an increase in PDH-E2. This shift in protein expression redirected pyruvate metabolism away from gluconeogenesis. SIRT3 overexpression reversed the suppressive effects of GZRG on gluconeogenesis, whereas SIRT3 knockdown synergized with GZRG. GZRG alleviates T2DM by inhibiting hepatic gluconeogenesis through the SIRT3-MPC1-PC/PDH axis. These findings elucidate a novel molecular mechanism of GZRG and highlight SIRT3 as a potential therapeutic target for T2DM management.

Introduction

Type 2 diabetes mellitus (T2DM) has emerged as one of the most prevalent and debilitating chronic metabolic disorders worldwide, posing a substantial public health and economic burden^1,2. Characterized by insulin resistance (IR) and progressive pancreatic β-cell dysfunction, T2DM is primarily driven by impaired glucose and lipid homeostasis^3,4. Among the multiple organs involved in glucose regulation, the liver plays a central role by controlling endogenous glucose production through glycogenolysis and gluconeogenesis⁵. Excessive hepatic gluconeogenesis is a major contributor to fasting hyperglycemia, a hallmark of T2DM pathophysiology, and represents a crucial therapeutic target for restoring glucose balance⁶.

Current hypoglycemic agents -- including metformin⁷, thiazolidinediones⁸, GLP-1 receptor agonists⁹, and SGLT2 inhibitors¹⁰ -- achieve glycemic control through distinct mechanisms, yet their long-term use is often limited by adverse effects, declining efficacy, or treatment intolerance. Consequently, there is increasing interest in complementary or alternative therapeutic approaches that can safely and effectively target multiple metabolic pathways. Traditional Chinese medicine (TCM), characterized by its multi-component and multi-target pharmacological properties, offers promising prospects for managing complex metabolic diseases such as T2DM^{11,12,13,14,15}. Several classical TCM formulations have been reported to ameliorate insulin resistance, improve lipid metabolism, and suppress hepatic gluconeogenesis through integrated regulatory mechanisms^16,17.

Ganzhirong granule (GZRG) is a standardized TCM formula with a history of clinical use in managing metabolic disorders. GZRG is an herbal formulation consisting of eight medicinal herbs. Based on our previous phytochemical and pharmacological investigations, a range of bioactive compounds has been identified in GZRG, including but not limited to Salvianolic acid B, D-stachyose, Proanthocyanidins, Violet oxalic acid, Salvianolic acid A, Salvianolic acid C, Calycosin 7 glucoside, 3′-methoxypuerarin, 3′-hydroxy puerarin, Apigenin-7-o-beta-d-pyrano, Emodin-8-O-β-D-glucose, Chrysophanol-1-O-glucoside, Chrysophanol-8-O-glucoside, Puerarin, Rosmarinic acid, Cryptotanshinone, Catechin, Ft-nyflavin, Calycosin, Rhein, Guanosine, Oleic acid, Aloe emodin, Formononetin, Palmitic acid, Chrysophanol, D-tryptophan, Danshensu, Ferulic acid, Mannitol, Gallic acid, and L-phenylalanine. A fundamental advantage of traditional Chinese medicine (TCM) in managing type 2 diabetes lies in its multi-component and multi-target synergistic actions. The dozens of active constituents contained in GZRG are likely to interact with multiple biological processes-such as insulin signaling pathways, glycogen metabolism, and inflammatory responses-thereby producing an integrated network effect that cannot be adequately replicated by any single purified compound¹⁸. Previous research has demonstrated that GZRG exerts beneficial effects on glucose and lipid metabolism in diabetic models¹⁹; however, the precise molecular mechanisms underlying these effects remain unclear. In particular, whether GZRG exerts its antidiabetic actions by modulating hepatic gluconeogenic signaling has not yet been elucidated.

Sirtuin 3 (SIRT3), a mitochondrial NAD⁺-dependent deacetylase, is a pivotal regulator of cellular energy homeostasis²⁰. By deacetylating and activating key metabolic enzymes, SIRT3 influences mitochondrial oxidative phosphorylation, fatty acid oxidation, and gluconeogenesis²¹. Aberrant SIRT3 expression has been implicated in the pathogenesis of insulin resistance, fatty liver, and T2DM^22,23. Evidence indicates that hepatic SIRT3 overexpression enhances gluconeogenic flux by modulating enzymes such as mitochondrial pyruvate carrier 1 (MPC1)²⁴, pyruvate carboxylase (PC)²⁵, pyruvate dehydrogenase (PDH)²⁶, phosphoenolpyruvate carboxykinase (PCK1)²⁷, and glucose-6-phosphatase (G6Pase)²⁸. Thus, SIRT3-mediated control of pyruvate metabolism represents a critical nexus linking mitochondrial function and hepatic glucose output, making SIRT3 an attractive target for therapeutic intervention in T2DM.

Given the established metabolic effects of bioactive constituents in GZRG, along with the central role of SIRT3 in hepatic glucose homeostasis, it was hypothesized that GZRG ameliorates hyperglycemia in T2DM through modulation of the SIRT3-mediated gluconeogenic pathway. To evaluate this hypothesis, the effects of GZRG on glucose and lipid metabolism were assessed in high-fat diet (HFD)-induced T2DM mice and free fatty acid (FFA)-induced insulin-resistant HepG2 cells. Further investigation examined the involvement of the SIRT3-MPC1-PC/PDH signaling axis in mediating the suppressive effect of GZRG on hepatic gluconeogenesis. This study aims to elucidate the molecular mechanisms underpinning the metabolic benefits of GZRG and to provide experimental evidence supporting its therapeutic potential as a multi-target modulator for T2DM management.

Protocol

The animal study protocol was approved by the Institutional Animal Care and Use Committee of Changchun University of Chinese Medicine (approval no. 2024396). All animal experiments were conducted following the national guidelines and the relevant national laws on the protection of animals.

Experimental drugs
GZRG (batch number 26210483) was provided by the Affiliated Hospital of Changchun University of Chinese Medicine (Jilin, China). The major bioactive constituents and quality control parameters of GZRG were characterized in a previous study¹⁸.

Reagents and antibodies
Commercial assay kits were used to quantify glucose, glycogen, non-esterified fatty acids (NEFAs), triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol. Enzyme-linked immunosorbent assay (ELISA) kits for glycosylated hemoglobin and insulin were also obtained commercially. ELISA kits for pyruvate dehydrogenase (PDH) and pyruvate carboxylase (PC) were also obtained commercially. For RNA and cDNA work, the RNA Tissue/Cell Kit, qPCR PreMix, and gDNA RT SuperMix were acquired commercially. Transfection was performed using a transfection reagent. Primary antibodies against SIRT3 (RRID: AB_2837621), phosphoenolpyruvate carboxykinase 1 (PCK1; RRID: AB_2838732), PDH-E2, glucose-6-phosphatase (G6Pase; RRID: AB_2845572), mitochondrial pyruvate carrier 1 (MPC1; RRID: AB_2836209), PC (RRID: AB_2836732), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; RRID: AB_2833041) were procured from a commercial vendor.

Mice and treatments
Male C57BL/6J mice (5-6 weeks old) were obtained from Liaoning Changsheng Biotechnology Co., Ltd. and maintained under standard conditions (22 ± 2 °C, 50%-60% humidity, 12 h light/dark cycle)with free access to food and water. Following a 1-week acclimatization period, six mice were randomly designated as the control (Con) group and fed a standard chow diet. The remaining mice received a high-fat diet (HFD) for 12 weeks to induce obesity and insulin resistance.

At week 13, mice exhibiting a fasting blood glucose (FBG) level ≥ 11.1 mmol/L were classified as successful type 2 diabetes mellitus (T2DM) models. These diabetic mice were randomly allocated into five experimental groups (n = 6 per group): the HFD model group, low-dose GZRG (L-GZRG, 3.2 g/kg/d), medium-dose GZRG (M-GZRG, 6.4 g/kg/d), high-dose GZRG (H-GZRG, 12.8 g/kg/d), and metformin (Met, 0.2 g/kg/d). GZRG and Met were administered daily by oral gavage for 8 weeks. The Con and HFD model groups received equivalent volumes of distilled water. Upon completion of the treatment period, blood and liver tissue samples were collected from all mice for subsequent analysis.

Intraperitoneal glucose tolerance test (IPGTT), pyruvate tolerance test (PTT), and insulin tolerance test (ITT)
Metabolic tolerance tests were performed between 6 and 8 weeks after the initiation of drug treatments. For the IPGTT and PTT, mice were fasted overnight (approximately 16 h). For the ITT, a shorter 4 h fasting period was implemented. Following fasting, mice received an intraperitoneal injection of either glucose (2 g/kg for IPGTT), sodium pyruvate (2 g/kg for PTT), or human regular insulin (1 U/kg for ITT). Blood glucose levels were measured from tail vein blood samples at 0 min, 30 min, 60 min, 90 min, and 120 min post-injection using a calibrated glucometer.

Biochemical analysis and enzyme activity assays
Fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and insulin (INS) levels were determined using commercial assay kits according to the manufacturers' instructions. The Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) was calculated using the formula:

HOMA-IR = [FBG (mmol/L) x INS (µU/mL)] / 22.5

Circulating levels of NEFAs, TG, TC, LDL-C, HDL-C, and the enzymatic activities of PDH and PC in liver tissues were quantified using the respective commercial kits.

Histopathological analysis of liver tissues
Fresh liver tissues were fixed in 10% neutral-buffered formalin for 24 h, followed by standard processing for paraffin embedding. Sections (5 µm thick) were cut and stained with Hematoxylin and Eosin (H&E) for general morphological assessment. For lipid visualization, select liver specimens were embedded in Optimal Cutting Temperature (OCT) compound, snap-frozen, cryosectioned, and stained with Oil Red O.

Cell culture and treatment
The human hepatoma cell line HepG2 was obtained from the Type Culture Collection of the Chinese Academy of Sciences (China). Cells were cultured in Minimum Essential Medium (MEM) supplemented with 25 mM glucose, 10% fetal bovine serum (FBS), and 1% penicillin-streptomycin. Cultures were maintained at 37 °C in a humidified incubator with 5% CO₂.

To establish an in vitro model of insulin resistance (IR), cells at 70%-80% confluence were treated for 24 h with a free fatty acid (FFA) mixture comprising oleic acid and palmitic acid at a 2:1 molar ratio, based on a previously reported method with minor modifications²⁹. Following FFA induction, the culture medium was replaced with fresh MEM containing the same FFA mixture along with various concentrations of GZRG for an additional 24 h incubation.

Cell viability assay (CCK-8)
HepG2 cells in the logarithmic growth phase were harvested and seeded into 96-well plates at a density of 1 x 10⁴ cells per well. The peripheral wells were filled with 100 µL of sterile phosphate-buffered saline (PBS) to minimize evaporation. After overnight incubation to ensure adherence, cells were treated according to the experimental design. Following treatment, 10 µL of CCK-8 solution was added to each well, and the plates were incubated for 2 h at 37 °C. The absorbance at 450 nm was measured using a microplate reader. Cell viability was calculated as:

[(As - Ab) / (Ac - Ab)] x 100%

where As is the absorbance of the sample, Ac is the absorbance of the control (untreated cells), and Ab is the absorbance of the blank (wells without cells).

Plasmid transfection in HepG2 cells
The SIRT3 overexpression plasmid (OV-SIRT3) and three distinct SIRT3-targeting small interfering RNA plasmids (si-SIRT3-1#, si-SIRT3-2#, si-SIRT3-3#), along with a negative control siRNA (si-NC), were constructed by a vendor. Transfections were performed using a transfection reagent when cells reached 50%-60% confluence. The transfection complex was prepared under sterile conditions as follows: Solution A was generated by diluting 1 µg of plasmid DNA in 250 µL of reduced serum medium with gentle mixing. Solution B was prepared by diluting 2-3 µL of transfection reagent in 250 µL of reduced serum medium, followed by incubation at room temperature for 5 min without vortexing. The two solutions were combined, mixed gently by pipetting, and incubated at room temperature for 30 min to form DNA-lipid complexes. For transfection, 1 h prior to the procedure, the cell culture medium was aspirated, cells were washed with PBS, and 1 mL of reduced serum medium (serum-free and antibiotic-free) was added. Then, 500 µL of the transfection complex was added dropwise to each well, and the culture plate was gently swirled to ensure homogeneity. Cells were incubated at 37 °C for 4-6 h, after which the medium was replaced with complete medium to mitigate cytotoxicity. Subsequently, cells were cultured for an additional 24-48 h before further analysis. Transfection efficiency was assessed by quantitative PCR (qPCR). Total RNA was extracted, reverse-transcribed into cDNA, and amplified using SIRT3-specific primers with 18S rRNA as an internal control. Relative expression levels were determined via the ΔΔCt method, and the most effective siRNA construct was selected for subsequent experiments based on these results.

Oil red O staining for intracellular lipid accumulation
HepG2 cells in the logarithmic growth phase were seeded in 24-well plates at a density of 2 x 10⁵ cells per well and induced with a high-lipid medium containing 250 µM oleic acid and 250 µM palmitic acid for 48 h to establish the lipid accumulation model. After treatments, cells were washed with PBS and fixed with 10% formaldehyde for 10 min at room temperature. Following a rinse with distilled water and a brief immersion in 60% isopropanol, the cells were stained with a freshly prepared Oil Red O working solution (obtained by mixing components A1 and A2 in a 3:2 ratio) for 10-15 min. Excess dye was removed by washing with 60% isopropanol, followed by distilled water. Nuclei were counterstained with Mayer's hematoxylin (1.0 g/L) for 1-2 min. After a final wash, cells were mounted with glycerol gelatin, and images were captured using a light microscope at 200x magnification for qualitative assessment of lipid droplets.

Induction of insulin resistance and glucose quantification in HepG2 cells
HepG2 cells were cultured in 24-well plates at a seeding density of 2 x 10⁵ cells per well to establish an in vitro model of insulin resistance. After 24 h of treatment, the culture supernatant was collected for further analysis. For intracellular glucose measurement, cell suspensions were centrifuged at 1000 x g for 10 min at 4 °C. The supernatant was discarded, and the cell pellet was washed once or twice with an isotonic buffer, followed by centrifugation under the same conditions. The pellet was then homogenized in 0.2 to 0.3 mL of homogenization medium and subjected to ultrasonic disruption in an ice-water bath using a 300 W probe, with pulses of 3-5 s followed by 30 s intervals, repeated 3x-5x. The resulting homogenate was assayed directly without further centrifugation. Glucose levels in both the culture supernatant and the cell homogenate were measured using a standardized glucose assay kit according to the manufacturer's instructions to ensure accurate and reproducible assessment of metabolic responses.

Measurement of cellular triglyceride content
HepG2 cells were collected by centrifugation at 1000 x g for 10 min, and the supernatant was discarded. The cell pellet was washed 1x or 2x with an isotonic phosphate buffer (0.1 M, pH 7.0-7.4), followed by centrifugation under the same conditions. After removal of the supernatant, the pellet was resuspended and homogenized with 0.2-0.3 mL of absolute ethanol. The homogenate was subjected to ultrasonic disruption on ice (300 W, 3-5 s per burst, 30 s intervals, repeated 3-5 times). The resulting homogenate was used directly for subsequent analysis without further centrifugation. A 96-well plate was set up with blank, standard, and sample wells. After thorough mixing, the plate was incubated at 37 °C for 10 min, and the absorbance was measured at 500 nm using a microplate reader. The triglyceride content in the cell samples was calculated based on a standard curve derived from serial dilutions of glycerol standards.

Measurement of total cholesterol content
HepG2 cells (5 x 10⁶) were lysed in 1 mL of extraction buffer by ultrasonic disruption on ice. The homogenate was centrifuged at 10,000 x g for 10 min at 4 °C, and the supernatant was collected for immediate analysis. A cholesterol standard stock solution (50 µmol/mL) was prepared and serially diluted to generate a standard curve (0.3125 to 10 µmol/mL). A working chromogenic solution was prepared by mixing reagents one, two, and three provided in the cholesterol assay kit, and pre-warmed to 37 °C. Samples or standards were mixed with the working solution, incubated at 37 °C for 30 min, and the absorbance was measured at 500 nm. Cholesterol content was calculated based on the standard curve.

Immunofluorescence staining
Cells grown on coverslips were rinsed with ice-cold PBS and fixed with 4% paraformaldehyde for 15 min at room temperature. After PBS washes, cells were permeabilized with 0.1% Triton X-100 for 10 min and blocked with 5% BSA at 4 °C overnight. Cells were then incubated with a primary anti-SIRT3 antibody (1:500 dilution) at 4 °C overnight. Following PBS washes, cells were incubated with a fluorescence-conjugated secondary antibody (1:2000 dilution) for 1 h at room temperature in the dark. Nuclei were stained with DAPI (0.5 µg/mL) for 10 min. Coverslips were mounted onto glass slides using an anti-fade mounting medium and visualized under a fluorescence microscope.

RNA extraction and quantitative real-time PCR (qRT-PCR)
Total RNA was extracted from cells using the RNA Tissue/Cell Kit. Genomic DNA was removed, and cDNA was synthesized using the gDNA RT kit. Quantitative PCR was performed using the qPCR kit on a real-time PCR detection system. Gene-specific primer sequences are provided in Table 1.

Western blot analysis
For protein extraction from liver tissues, approximately 50-200 mg of tissue was weighed, minced in a petri dish, and transferred to a homogenizer. Pre-cooled lysis buffer (RIPA buffer) supplemented with protease and phosphatase inhibitors was added to prevent protein degradation. Homogenization was performed thoroughly on ice to ensure complete tissue disruption, and the homogenate was transferred to a centrifuge tube and incubated on ice for 30 min to facilitate protein solubilization. Subsequently, the homogenate was centrifuged at 4 °C at 12,000 x g for 10-30 min, and the supernatant was carefully collected as the total protein extract, avoiding contamination from the pellet. For cellular protein extraction, pre-cooled lysis buffer (RIPA buffer containing 1% Triton X-100 and protease inhibitors) was added to the cells, followed by incubation on ice for 15-30 min to achieve complete lysis. Cell disruption was ensured by repeated scraping with a cell scraper or vigorous pipetting. The lysate was transferred to a centrifuge tube and centrifuged at 4 °C at 14,000 x g for 30 min, and the supernatant was collected as the total protein extract. Protein concentration was quantified using a BCA protein assay kit, and aliquots were stored at -80 °C for future Western blot analysis or used directly for sample preparation. Protein samples were denatured in Laemmli buffer at 100 °C for 10 min, separated by SDS-PAGE, and transferred onto PVDF membranes. Membranes were blocked with 5% non-fat milk for 2 h at room temperature and subsequently incubated with specific primary antibodies overnight at 4 °C. After washing, membranes were incubated with HRP-conjugated secondary antibodies for 1 h at room temperature. Protein bands were visualized using an enhanced chemiluminescence (ECL) substrate. The chemiluminescent signals were captured, and band intensities were quantified by densitometric analysis using ImageJ software. The raw Western blots are provided in Supplementary File 1.

Statistical analysis
All data are expressed as the mean ± standard error of the mean (SEM). Statistical comparisons between two groups were performed using an unpaired Student's t-test. For comparisons among three or more groups, one-way analysis of variance (ANOVA) was applied, followed by Tukey's post hoc test for multiple comparisons. All statistical analyses were conducted using GraphPad Prism software (Version 9.5.0). A p-value of less than 0.05 was considered statistically significant.

Results

GZRG ameliorated insulin resistance and gluconeogenesis in HFD-induced T2DM mice
The HFD-induced type 2 diabetes mellitus (T2DM) mouse model, characterized by insulin resistance, hyperglycemia, and dyslipidemia, is widely employed to investigate disease pathogenesis and evaluate therapeutic interventions³⁰. In the present study, an HFD-induced T2DM model was established in C57BL/6J mice to assess the efficacy and underlying mechanisms of GZRG.

Compa...

Discussion

The present study demonstrates that GZRG exerts significant antihyperglycemic and insulin-sensitizing effects in both in vivo and in vitro models of T2DM, primarily through suppression of hepatic gluconeogenesis via modulation of the SIRT3-MPC1-PC/PDH axis. Experimental data indicate that GZRG administration attenuated fasting hyperglycemia, improved glucose and insulin tolerance, restored lipid homeostasis, and reduced hepatic steatosis in HFD-induced diabetic mice. In parallel, treatment with GZRG enh...

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
FastKing gDNA Dispelling RT SuperMix	Tiangen Biotech Co., Ltd. (China).	KR118-02
glucose-6-phosphatase (G6Pase) Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-D-12610
glyceraldehyde-3-phosphate dehydrogenase (GAPDH) Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-T-0004
Glycosylated hemoglobin ELISA kits	Jiangsu Meimian Industrial Co., Ltd. (China).	MM-0159M2
GZRG	Affiliated Hospital of Changchun University of Chinese Medicine (Jilin, China)	26210483
high-density lipoprotein (HDL) cholesterol assay kits	Nanjing Jiancheng Bioengineering Institute (China)	A112-1-1
insulin ELISA kits	Jiangsu Meimian Industrial Co., Ltd. (China).	MM-0579M2
Lipofectamine2000	Invitrogen, USA	11668-019
Low-density lipoprotein (LDL) cholesterol assay kits	Nanjing Jiancheng Bioengineering Institute (China)	A113-1-1
measure glucose assay kits	Nanjing Jiancheng Bioengineering Institute (China)	F006-1-1
mitochondrial pyruvate carrier 1 (MPC1)Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-D-3852
non-esterified fatty acids (NEFAs)assay kits	Nanjing Jiancheng Bioengineering Institute (China)	A042-2-1
PC Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-D-4364
PDH-E2 Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-D-14046
phosphoenolpyruvate carboxykinase 1 (PCK1)Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-D-6770
pyruvate carboxylase (PC) ELISA kits	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-160408H
Pyruvate dehydrogenase (PDH)ELISA kits	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-160300H
RNA Easy Fast Tissue/Cell Kit	Tiangen Biotech Co., Ltd. (China).	DP451
SIRT3 Antibody	Wuhan Lingjiesi Biotechnology Co., Ltd. (China)	LJS-A-5135
Talent SYBR Green qPCR PreMix	Tiangen Biotech Co., Ltd. (China).	FP209-02
total cholesterol (TC)assay kits	Nanjing Jiancheng Bioengineering Institute (China)	A110-1-1
triglycerides (TG)assay kits	Nanjing Jiancheng Bioengineering Institute (China)	A111-1-1