Protective Effect of Naoxintong Capsule () Combined with Guhong Injection () on Rat Brain Microvascular Endothelial Cells during Cerebral Ischemia-Reperfusion Injury / 中国结合医学杂志

OBJECTIVE@#To investigate the synergistic effect of Naoxintong Capsule (NXTC, ) and Guhong Injection (GHI, ) on cerebral ischemia-reperfusion (I/R) injury.@*METHODS@#Forty-eight Sprague-Dawley rats were divided into 6 groups: control group, oxygen and glucose deprivation (OGD) group, nimodipine group (9.375 mg/kg), NXTC group (0.5 g/kg), GHI group (5 mL/kg) and NXTC+GHI group (0.5 g/kg NXTC+5 mL/kg GHI), after the onset of reperfusion and once per day for the following 7 days. Blood was collected 1 h after final administration, and the sera were collected. Cultured primary rat brain microvascular endothelial cells (rBMECs) were subjected to OGD to establish a cell injury model. Untreated rBMECs were used as blank control. The cell counting kit-8 assay was used to assess cell viability using the sera. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were assessed using an enzyme-linked immunosorbent assay. Apoptosis was evaluated after Hoechst33342 staining using fluorescence microscopy and flow cytometry. JC-1 staining was performed to assess changes in mitochondrial membrane potential.@*RESULTS@#Statistical analysis indicated that more than 95% of the cells were rBMECs. Compared with the OGD group, the cellular morphology of the all drug delivery groups improved. In particular, the combined drug group had the most significant effect. Compared with the OGD group, all drug intervention groups induced a decrease in the apoptotic rate of rBMECs, increased the SOD levels, and decreased the MDA levels (all P<0.01). Compared with the mono-therapy groups, the NXTC+GHI group exhibited a significant improvement in the number of apoptotic rBMECs (P<0.01). All drug intervention groups showed different degrees of increase in membrane potential, and the NXTC+GHI group was higher than the NXTC or GHI group (P<0.01).@*CONCLUSION@#The combinationa application of NXTC and GHI on cerebral I/R injury clearly resulted in protective benefits.

Mechanism of Guhong Injection against Hypoxia/Reoxygenation Injury of H9c2 Cardiomyocytes Based on PI3K/AKT Signaling Pathway / 中国药学杂志

OBJECTIVE: To explore the protectiveness of Guhong injection (GHI) on H9c2 cardiomyocytes injured by hypoxia/reoxygenation (H/R) based on PI3K/AKT signaling pathway. METHODS: H9c2 cardiomyocytes were cultured and the optimal GHI doses were screened by CCK-8. The cells were randomly subjected into 8 groups: control group, model group, low, medium and high GHI dose groups (30, 60 and 90 μL•mL-1 respectively), positive drug group (verapamil injection 7.5 μL•mL-1), GHI+LY294002 (PI3K inhibitor) group and LY294002 group. Cells were established H/R model with hypoxia for 4 h and reoxygenation for 16 h excepted the control group. The CK-MB, LDH, MDA content and SOD activities in each group were detected by Elisa, and the expression levels of p-PI3K, PI3K, p-AKT, AKT and GSK-3β in each group were detected by Western blot. RESULTS: Compared each drug-treated group with the model group, LDH and CK-MB contents were decreased (P<0.05), MDA content was decreased (P<0.01), and SOD activity was increased (P<0.01). Meanwhile, the expression levels of p-PI3K/PI3K, p-AKT/AKT, and GSK-3β protein were elevated (P<0.01). Compared with the model group, LDH, CK-MB, MDA and SOD activity in LY294002 group were no significant difference, while the expression levels of p-PI3K/PI3K, p-AKT/AKT and GSK-3β protein decreased (P<0.01). CONCLUSION: GHI could represent anti-oxidative and anti-apoptotic effect which reduced the damage of injured H9c2 myocardial cells caused by H/R. These effects might be related to the PI3K/AKT signaling pathway.

Mechanism of Guhong Injection Against Cerebral Ischemia Based on Transcriptome Analysis / 中国实验方剂学杂志

Objective::Based on the protective effect of Guhong injection (GH) on cerebral ischemia, mechanism of GH against cerebral ischemia was identified using RNA-seq transcriptome and bioinformation analysis. Method::The model of middle cerebral artery occlusion (MCAO) was established through thread embolization. Sham group, model group, low-dose GH group (0.625 mL·kg-1·d-1), high-dose GH group (2.5 mL·kg-1·d-1), positive group (Ginaton, 8 mL·kg-1·d-1) were set up. Ludmila Belayev 12-point scoring method was applied to assess the protective effect of GH against MCAO-induced cerebral ischemia. And the differentially expressed genes after treatment with GH were identified by RNA-Seq technology. Enrichment analysis, cluster analysis and association analysis on disease targets of cerebral ischemia were carried out through such databases as DAVID, String and The Human Phenotype Ontology. Finally, the regulatory network was constructed by Cytoscape3.4.0. Result::Compared with the sham group, the neurological impairment was obvious in the model group (P<0.01), and the neurological impairment was alleviated in the GH group compared with the model group (P<0.05). RNA-Seq technology analysis showed that GH regulated genes involving such biological processes as cell apoptosis, inflammation, oxidative stress, toll-like signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway. Twenty disease targets and 64 MAPK signaling pathway genes were associated with differentially expressed genes after GH treatment, in which 23 genes were involved in apoptosis and inflammation. Conclusion::GH protected against cerebral ischemia in many ways, among which MAPK signaling pathway is an important way to exert its effect in inhibiting apoptosis and inflammation.

Pharmacokinetics of main activ ingredients of Guhong injection in cerebral ischemia rats / 中国中药杂志

To investigate the pharmacokinetic characteristics of active constituents of Guhong injection in rats with cerebral ischemia reperfusion injury. The middle cerebral artery occlusion (MCAO) model was established in our studies, and then all the rats received iv administration of Guhong injection (2.1 mL·kg⁻¹). The blood concentrations of aceglutamide and hydroxysafflor yellow A (HSYA) were determined by high performance liquid chromatography (HPLC) method at different time points. The concentration-time curves were drawn and pharmacokinetic data were obtained by DAS 3.2.6 software. The results showed that aceglutamide and HSYA showed good linear relationship within the ranges of 1.5-500 mg·L⁻¹ (R²=0.997 5) and 0.33-40 mg·L⁻¹ (R²=0.998 9) respectively. This quantitative method showed a high recovery rate, good precision and stability. The main pharmacokinetics parameters of t1/2α, t1/2β, CL₁, CL₂, AUC0-t, AUC0-∞, Vd1, and Vd2 were (0.139±0.007) and (0.155±0.017) h, (0.803±0.046) and (2.233±0.410) h, (0.016±0) and (0.149±0.018) L·h⁻¹·kg⁻¹, (0.015±0.001) and (0.446±0.016) L·h⁻¹·kg⁻¹, (133.335±3.844) and (9.298±0.179) mg·h·L⁻¹, (143.851±3.595) and (14.464±1.451) mg·h·L⁻¹, (0.009±0.001) and (0.223±0.007) L·kg⁻¹, (0.006±0.001) and (0.212±0.032) L·kg⁻¹, respectively. The results showed that the established HPLC method was highly specific, and could be used for the simultaneous detection of aceglutamide and HSYA of Guhong injection in MCAO rats, which was conducive to pharmacokinetic studies. Pharmacokinetic data and parameters could provide reference for continuous administration and interval administration of the drug.

Effect and Mechanism of Guhong Injection Promoting Fracture Healing in Rats / 中国药学杂志

OBJECTIVE: To observe the effect of guhong injection(GHI) on tibial fracture healing in rats and to explore the mechanism of the action of GHI. METHODS: One hundred and eighty male SD rats were randomly divided into 6 groups with 30 rats in each group: sham operation group, model group, positive drug group(compound ossotide injection, 5 mL•kg-1), low, medium and high dose of GHI groups(2.5, 5, 10 mL•kg-1). In addition to the sham operation group, the other groups established the rat model of tibial fracture. All were given once daily intraperitoneal injections and samples were taken at 1st, 2nd, 4th and 6th week. Blood biochemical analysis and Elisa kit detection were performed on blood samples. X-rays, biomechanical tests, immunohistochemistry and RT-PCR were performed on the tibial samples. RESULTS: ①After administration for one, two and four weeks, the levels of serum calcium(Ca) and phosphorus(P) in medium and high dose of GHI groups were higher than those in model group(P<0.05). ②X-ray showed that the outer callus growth and the disappearance of fracture line in all dose groups of GHI were faster than those in model group. ③Compared with model group, the maximum load and rigidity of medium and high dose of GHI groups were increased at each time point(P<0.05), and the trend of stress line graph were improved obviously. The content of alkaline phosphatase(ALP) in medium and high dose of GHI groups were higher than that in model group at each time point(P<0.05). Compared with model group, the serum levels of PDGF were increased in all dose groups of GHI(P<0.05 or P<0.01). After administration for one, two and four weeks, the serum BMP-2 in all dose groups of GHI were higher than those in model group(P<0.05 or P<0.01). ⑤Compared with model group, the expression of Runx2 mRNA were increased in medium and high dose of GHI groups, as well as Smad5 protein expression(P<0.05 or P<0.01). CONCLUSION: GHI could significantly improve the biomechanical properties of bone in fracture rats. The promotion of fracture healing might be through the upregulation of PDGF and BMP-2 expression in different stages of bone healing, and the regulation of BMP/Smad5/ Runx2 signaling may be one of the mechanisms of promoting fracture healing.

Effects of Guhong injection on ATPase activity and inflammatory response after cerebral ischemia/reperfusion injury in rats / 中国中西医结合急救杂志

Objective To investigate the influence of Guhong injection (GHI) on ATPase activity and inflammatory response after cerebral ischemia/reperfusion (I/R) injury in rats, and evaluate its protective effects on cerebral I/R injury. Methods Seventy-two male Sprague-Dawley (SD) rats were divided into sham group, I/R group, nimodipine group, and the low-dose (2.5 mL/kg, GHI-L), medium-dose (5.0 mL/kg, GHI-M), and high-dose (10.0 mL/kg, GHI-H) of GHI groups according to the random number table method, with 12 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by the intraluminal suture method to prepare the model of focal cerebral ischemia, and reperfusion was performed after 1.5 hours of occluding the middle cerebral artery; the sham group had the same operation process except inserting the nylon thread. The injection of drug in various drug-treated groups was carried out via a tail vein at 0, 12, 24 hours after the onset of reperfusion, while the sham group and I/R group received the same amount of normal saline. At 12 hours after last drug administration, the scores of neurological deficit symptoms were evaluated; the cerebral infarction was observed by triphenyl tetrazolium chloride (TTC) staining; the Na+-K+-ATPase and Ca2+-ATPase activities in the brain tissue were measured by phosphorus determination; the contents of interleukin-6 (IL-6), monocyte chemotactic factor-1 (MCP-1), nitric oxide (NO) in serum were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the sham group, the neurological function score was significantly decreased, the cerebral infarction was serious, the activities of ATPase was obviously decreased, and the levels of serum inflammatory factors were significantly increased in I/R group. Compared with the I/R group, the neurological function scores were significantly increased in GHI-L group, GHI-M group, GHI-H group and nimodipine group (9.03±0.63, 10.54±2.55, 12.33±1.87, 12.06±1.89 vs. 8.17±1.05, all P < 0.05), the volumes of cerebral infarction were obviously reduced [(18.51±1.80)%, (15.98±1.34)%, (8.61±1.16)%, (8.09±0.96)% vs. (26.52±2.07)%, all P < 0.01], the activities of ATPase were significantly increased [Na+-K+-ATPase (μmol·mg-1·h-1):5.10±0.30, 5.34±0.26, 6.19±0.17, 5.86±0.31 vs. 3.98±0.35, Ca2+-ATPase (μmol·mg-1·h-1): 3.68±0.44, 4.43±0.29, 5.03±0.27, 4.17±0.30 vs. 1.87±0.46, all P < 0.01], and the levels of serum inflammatory factors were decreased obviously [IL-6 (ng/L): 51.61±5.55, 43.88±4.05, 39.71±2.22, 41.28±2.66 vs. 60.11±6.61, MCP-1 (ng/L): 227.82±7.07, 201.58±13.10, 177.23±10.46, 126.80±8.49 vs. 296.01±12.85, NO (μmol/L): 54.48±3.23, 46.84±2.69, 41.15±2.80, 48.62±2.34 vs. 65.25±3.88, all P < 0.05]. Conclusions GHI not only can improve the energy metabolism of brain tissue in a dose-dependent manner, but also inhibit the inflammatory cascade of damage after cerebral I/R in rats, which might be its protective mechanism on cerebral ischemia injury.

Effects of Guhong injection on ATPase activity and inflammatory response after cerebral ischemia/reperfusion injury in rats / 中国中西医结合急救杂志

Objective To investigate the influence of Guhong injection (GHI) on ATPase activity and inflammatory response after cerebral ischemia/reperfusion (I/R) injury in rats, and evaluate its protective effects on cerebral I/R injury. Methods Seventy-two male Sprague-Dawley (SD) rats were divided into sham group, I/R group, nimodipine group, and the low-dose (2.5 mL/kg, GHI-L), medium-dose (5.0 mL/kg, GHI-M), and high-dose (10.0 mL/kg, GHI-H) of GHI groups according to the random number table method, with 12 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by the intraluminal suture method to prepare the model of focal cerebral ischemia, and reperfusion was performed after 1.5 hours of occluding the middle cerebral artery; the sham group had the same operation process except inserting the nylon thread. The injection of drug in various drug-treated groups was carried out via a tail vein at 0, 12, 24 hours after the onset of reperfusion, while the sham group and I/R group received the same amount of normal saline. At 12 hours after last drug administration, the scores of neurological deficit symptoms were evaluated; the cerebral infarction was observed by triphenyl tetrazolium chloride (TTC) staining; the Na+-K+-ATPase and Ca2+-ATPase activities in the brain tissue were measured by phosphorus determination; the contents of interleukin-6 (IL-6), monocyte chemotactic factor-1 (MCP-1), nitric oxide (NO) in serum were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the sham group, the neurological function score was significantly decreased, the cerebral infarction was serious, the activities of ATPase was obviously decreased, and the levels of serum inflammatory factors were significantly increased in I/R group. Compared with the I/R group, the neurological function scores were significantly increased in GHI-L group, GHI-M group, GHI-H group and nimodipine group (9.03±0.63, 10.54±2.55, 12.33±1.87, 12.06±1.89 vs. 8.17±1.05, all P < 0.05), the volumes of cerebral infarction were obviously reduced [(18.51±1.80)%, (15.98±1.34)%, (8.61±1.16)%, (8.09±0.96)% vs. (26.52±2.07)%, all P < 0.01], the activities of ATPase were significantly increased [Na+-K+-ATPase (μmol·mg-1·h-1):5.10±0.30, 5.34±0.26, 6.19±0.17, 5.86±0.31 vs. 3.98±0.35, Ca2+-ATPase (μmol·mg-1·h-1): 3.68±0.44, 4.43±0.29, 5.03±0.27, 4.17±0.30 vs. 1.87±0.46, all P < 0.01], and the levels of serum inflammatory factors were decreased obviously [IL-6 (ng/L): 51.61±5.55, 43.88±4.05, 39.71±2.22, 41.28±2.66 vs. 60.11±6.61, MCP-1 (ng/L): 227.82±7.07, 201.58±13.10, 177.23±10.46, 126.80±8.49 vs. 296.01±12.85, NO (μmol/L): 54.48±3.23, 46.84±2.69, 41.15±2.80, 48.62±2.34 vs. 65.25±3.88, all P < 0.05]. Conclusions GHI not only can improve the energy metabolism of brain tissue in a dose-dependent manner, but also inhibit the inflammatory cascade of damage after cerebral I/R in rats, which might be its protective mechanism on cerebral ischemia injury.

Study on the Stability of Guhong Injection in Infusions Commonly Used in Clinics / 中国药师

Objective:To investigate the stability of Guhong injection in infusions commonly used in clinics. Methods:The chan-ges in pH value, appearance and insoluble particles were observed, and the contents of aceglutamide, uridine, adenosine, guanosine, lilac glycosides, hydroxyl safflower yellow prime A and anhydrosafflor yellow B in Guhong injection were determined by HPLC-DAD in 8 h after the compatibility respectively at 5, 25 and 35℃ under the condition of avoiding light, indoor illumination and ultraviolet irra-diation, respectively. The contents of the related substances of acetyl glutamine were detected by HPLC. Results:In 8 h, the appear-ance, pH and insoluble particles of all compatibility solution had no significant changes. The content of aceglutamide in compatibility solution was decreased under the condition of ultraviolet irradiation (35 ℃), and the content of compositions in the other compatibility solution showed no significant changes. Under the condition of avoiding light and indoor illumination, the content of relative substances showed no significant changes before and after the compatibility, and met the relevant provisions. But under ultraviolet light, with tem-perature increasing and time prolonging, the content of the related substances was increased significantly. Conclusion: Guhong injec-tion is stable in 5 % glucose injection, 10 % glucose injection and sodium chloride injection in 4 hours, while the mixtures should a-void sunlight during the use.

Correlation study on in vivo pharmacokinetics and anti-oxidation of Guhong Injection in cerebral ischemia reperfusion injury model of rats / 中草药

Objective: To investigate the in vivo pharmacokinetic progress of hydroxysafflor yellow A (HSYA) from Guhong Injection in cerebal ischemia reperfusion (I/R) injury of rats and the correlation with its anti-oxidation effect. Methods: The equilibrium dialysis method was carried out to determine the plasma protein binding rates of HYSA and HSYA in Guhong Injection. Middle cerebral artery occlusion (MCAO) model rats were iv injected HYSA (4 mg/kg) or Guhong Injection (10 mL/kg). The HPLC method was adopted to determine the plasma concentration of HYSA at different time points to draw the drug-time curve. Meanwhile, glutathione peroxidase (GSH-Px) and lactate dehydrogenase (LDH) activities were determined to draw the time-effect curve. Furthermore, the relationship between pharmacokinetics and pharmacodynamics was analyzed. Results: At the concentration of 2.5, 10, and 25 mg/L, the p plasma rotein binding rates of HYSA were 77.96%, 73.54%, and 76.13%, whereas the plasma protein binding rates of HYSA from Guhong Injection were 68.21%, 58.22%, and 63.17%, respectively. A good linear relationship of HYSA was obtained in the range of 0.01-50 mg/L, the mean recoveries were (99.94 ± 2.82)%, (104.16 ± 1.41)%, and (99.74 ± 1.06)% for low, middle, and high concentration of the samples, respectively. Compared with HYSA group, Guhong Injection significantly increased the AUC of HYSA and decreased the MRT and Vz of HYSA. Furthermore, Guhong Injection increased the content of GSH-Px and decreased the content of LDH. The plasma concentration of HYSA is positively related to the GSH-Px activity and negatively related to the LDH activity. Conclusion: The results indicate that HYSA has the moderate plasma protein binding rate. Compared with HYSA group, the plasma protein binding rate in Guhong Injection group is reduced. Guhong Injection could increase the bioavailability of HYSA to enhance therapeutic efficacy and increase the distribution of HYSA in ischemia rats. Guhong Injection has better anti-oxidant effect, as well as more significant protective effect against cerebral I/R injury than HYSA.

Effects of Guhong Injection on Expression of Vascular Endothelial Growth Factor in Cortex after Cerebral Isch-emia-reperfusion in Rats / 中国康复理论与实践

Objective To study the effects of Guhong injection on the expression of vascular endothelial growth factor (VEGF) in the cortex 14 days after cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were divided into sham group (n=6), ischemia group (n=6), aceglutamide injection group (n=6), Honghua injection group (n=6) and Guhong injection group (n=6). The middle cerebral ar-teries of all the rats were occluded for 2 hours and reperfusion, except the sham group. Drugs were administered once a day 24 hours after re-perfusion. The expression of VEGF in cortex was detected with enzyme-linked immunosorbent assay (ELISA) 14 days after reperfusion. Re-sults The expression of VEGF decreased in the ischemia group compared with the sham group (P<0.001), and it increased both in the ace-glutamide and Guhong injection groups compared with the ischemia group (P<0.05). Conclusion Guhong injection can significantly in-crease the expression of VEGF in the cortex 14 days after ischemia-reperfusion, which may be one of the ways for neuro-protection.

Effects of Guhong Injection on Motor Dysfunction in Cerebral Ischemia-reperfusion Rats / 中国康复理论与实践

Objective To evaluate the effects of Guhong Injection on motor dysfunction in rats after cerebral ischemia-reperfusion. Methods The middle cerebral arteries were occluded for 2 hours and re-perfused in Sprague-Dawley rats. They were divided in sham group, model group, Aceglutamide group, Safflowere group and Guhong group, which were intravenously administrated with normal saline, Ace-glutamide, Safflower or Guhong 24 hours after operation, and continued for 14 days. They were tested with the beam-walking test after treat-ment. Tyrosine hydroxylase (TH) immunohistochemical staining was used to investigate the viability of neurons in the substantia nigra. Re-sults The model group spent more time in the beam-walking test than that in the sham group (P<0.01), and it decreased in the Safflower group and Guhong group compared with that in the model group (P<0.05). The TH-positive neurons decreased in the model rat compared with that in the sham group (P<0.001), and increased in both Safflower and Guhong groups compared with that in the model group (P<0.01). Conclusion Guhong administration could significantly improve the motor dysfunction in rats after cerebral ischemia-reperfusion, which might be related to provent the neurons from injury in the substantia nigra.

Effects of Guhong Injection on Motor Dysfunction in Cerebral Ischemia-reperfusion Rats / 中国康复理论与实践

@# Objective To evaluate the effects of Guhong Injection on motor dysfunction in rats after cerebral ischemia- reperfusion. Methods The middle cerebral arteries were occluded for 2 hours and re-perfused in Sprague-Dawley rats. They were divided in sham group, model group, Aceglutamide group, Safflowere group and Guhong group, which were intravenously administrated with normal saline, Aceglutamide, Safflower or Guhong 24 hours after operation, and continued for 14 days. They were tested with the beam-walking test after treatment. Tyrosine hydroxylase (TH) immunohistochemical staining was used to investigate the viability of neurons in the substantia nigra. Results The model group spent more time in the beam-walking test than that in the sham group (P<0.01), and it decreased in the Safflower group and Guhong group compared with that in the model group (P<0.05). The TH-positive neurons decreased in the model rat compared with that in the sham group (P<0.001), and increased in both Safflower and Guhong groups compared with that in the model group (P<0.01). Conclusion Guhong administration could significantly improve the motor dysfunction in rats after cerebral ischemia- reperfusion, which might be related to provent the neurons from injury in the substantia nigra.

Effects of Guhong Injection on Expression of Vascular Endothelial Growth Factor in Cortex after Cerebral Ischemia-reperfusion in Rats / 中国康复理论与实践

@#Objective To study the effects of Guhong injection on the expression of vascular endothelial growth factor (VEGF) in the cortex 14 days after cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were divided into sham group (n=6), ischemia group (n=6), aceglutamide injection group (n=6), Honghua injection group (n=6) and Guhong injection group (n=6). The middle cerebral arteries of all the rats were occluded for 2 hours and reperfusion, except the sham group. Drugs were administered once a day 24 hours after reperfusion. The expression of VEGF in cortex was detected with enzyme-linked immunosorbent assay (ELISA) 14 days after reperfusion. Results The expression of VEGF decreased in the ischemia group compared with the sham group (P<0.001), and it increased both in the aceglutamide and Guhong injection groups compared with the ischemia group (P<0.05). Conclusion Guhong injection can significantly increase the expression of VEGF in the cortex 14 days after ischemia-reperfusion, which may be one of the ways for neuro-protection.

Determination of Total Polyphenols in Guhong Injection / 浙江中医药大学学报

Objective To establish determination method for total polyphenols in Guhong Injection, and provide reference for internal quality control of Guhong Injection.[Methods]Using the Folin-Ciocalteu colorimetric method, according to the consistent degree of the maximum absorption wavelengths of hydroxysafflor yel ow A and kaempferol relative to Guhong Injection, we selected the appropriate reference and color reaction condition to content determination.[Rsults]With hydroxysafflor yel ow A as the standard, the content of total polyphenols in 10 batches of samples were 9.94, 9.55, 9.75, 9.67, 9.84, 10.03, 9.81, 9.52, 9.88, 10.09mg·mL-1. The average recovery of total polyphenols was 98.11% and RSD was 1.68%(n=6). [Conclusion]The established Folin-Ciocalteu method is simple, accurate, sensitive, and is reliable for detecting total polyphenols in Guhong Injection.