Treatment of diabetic retinopathy with herbs for tonifying kidney and activating blood circulation: A review of pharmacological studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic retinopathy (DR) is a prevalent microvascular complication of diabetes. Chinese medicine believes that kidney deficiency and blood stasis are significant pathogenesis of DR. A characteristic therapeutic approach for this pathogenesis is the kidney-tonifying and blood-activating method. By literature retrieval from several databases, we methodically summarized the commonly used kidney-tonifying and blood-activating herbs for treating DR, including Lycii Fructus, Rehmanniane Radix Praeparata, and Corni Fructus with the function of nourishing kidney; Salvia Miltiorrhizae Radix et Rhizoma with the function of enhancing blood circulation; Rehmanniae Radix with the function of nourishing kidney yin; and Astragali Radix with the function of tonifying qi. It has been demonstrated that these Chinese herbs described above, by tonifying the kidney and activating blood circulation, significantly improve the course of DR. AIM OF THE STUDY: Through literature research, to gain a thorough comprehension of the pathogenesis of DR. Simultaneously, through the traditional application analysis, modern pharmacology research and network pharmacology analysis of kidney-tonifying and blood-activating herbs, to review the effectiveness and advantages of kidney-tonifying and blood-activating herbs in treating DR comprehensively. MATERIALS AND METHODS: PubMed, the China National Knowledge Infrastructure (CNKI), and Wanfang Data were used to filter the most popular herbs for tonifying kidney and activating blood in the treatment of DR. The search terms were "diabetic retinopathy" and "tonifying kidney and activating blood". Mostly from 2000 to 2023. Network pharmacology was applied to examine the key active components and forecast the mechanisms of kidney-tonifying and blood-activating herbs in the treatment of DR. RESULTS: Kidney deficiency and blood stasis are the pathogenesis of DR, and the pathogenesis is linked to oxidative stress, inflammation, hypoxia, and hyperglycemia. Scientific data and network pharmacology analysis have demonstrated the benefit of tonifying kidney and activating blood herbs in treating DR through several channels, multiple components, and multiple targets. CONCLUSIONS: This review first presents useful information for subsequent research into the material foundation and pharmacodynamics of herbs for tonifying kidney and activating blood, and offers fresh insights into the treatment of DR.

Exploring the mechanism of Bushen Huoxue prescription in the treatment of early diabetic retinal edema from the perspective of inner blood-retinal barrier injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic retinopathy (DR) is one of the most severe complications of diabetes mellitus, diabetes belongs to the category of "emaciation-thirst disease" in traditional Chinese medicine (TCM). Bushen Huoxue Prescription (BHP) is composed of traditional Chinese materia medica, which has therapeutic effects on DR and early diabetic retinal edema (EDRE). However, the therapeutic mechanism is unclear. AIM OF THE STUDY: Exploring the mechanism of BHP against EDRE. METHODS: Feeding Sprague Dawley (SD) rats a high-fat, high-sugar diet as well as providing intraperitoneal injections of streptozotocin (STZ) to promote inner blood-retinal barrier (iBRB) damage that can trigger EDRE, evaluating the therapeutic effect of BHP by the level of expressiveness of TJ proteins (ZO-1,Occludin) of the iBRB and the leakage of rhodamine B isothiocyanate (RITC) in the retina. The combination of network pharmacology and metabolomics was employed to study the mechanism of BHP in preventing of EDRE, then four proteins which were closely to the damage of iBRB were chosen for the validation by employing Western Blot (WB). RESULTS: Research of network pharmacology had shown that BHP had efficacy against EDRE by regulating targets such as AKT1, ALB, TNF, PPARG, etc, its potential pathways mainly involving signaling pathways such as HIF-1. In untargeted metabolomics analysis of serum, 15 differential metabolites were identified, with the metabolic pathways focusing on ketone body metabolism and synthesis, sphingolipid metabolism and phenylalanine metabolism. The conclusions of metabolomics and network pharmacology revealed that BHP can treat EDRE by alleviating hypoxia and oxidative stress and exerting protection of the iBRB. Finally, BHP's protection behavior of the iBRB was validated by WB experiments. CONCLUSION: Through integrating pharmacodynamics, network pharmacology and metabolomics, BHP was discovered to have a crucial function in EDRE therapy by preserving the integrity of iBRB. This comprehensive strategy also provided a reasonable way to reveal the multi-components, multi-targets, multi-pathways mechanism of TCM.

Exploring the quality markers and mechanism of Bushen Huoxue Prescription in prevention and treatment of diabetic retinopathy based on Chinmedomics strategy.

ETHNOPHARMACOLOGICAL RELEVANCE: Chinese herbal medicines have complex chemical composition; therefore, revealing the effective substances of Chinese herbal medicine becomes a prerequisite for scientific elucidation of the mechanism of action of Bushen Huoxue Prescription (BHP) against diabetic retinopathy (DR) and the development of new drugs. AIM OF THE STUDY: The Chinmedomics technique was used to evaluate the pharmacodynamic ingredients and mechanism of action of BHP against DR rats. MATERIALS AND METHODS: The overall physiological condition of the rats, including body weight, blood glucose, inflammatory factor levels, histological staining, and urine metabolic profile were examined to evaluate the model and its effects. The chemical composition of BHP in vivo and ex vivo was fully analyzed utilizing UPLC-Q-Exactive Orbitrap MS in conjunction with TCM serum pharmacochemistry. Finally, correlation analysis between biomarkers, and serum migration components was used to identify Quality markers (Q-markers) that were significantly associated with effectiveness. RESULTS: The UPLC-Q-Exactive Orbitrap MS platform was used to identify a total of 29 chemicals in serum, 17 of which were highly linked with effectiveness and can be potentially employed as pharmacodynamic substances for BHP against DR. In addition, 14 biomarkers related to galactose metabolism, starch and sucrose metabolism, pantothenate and CoA biosynthesis, glycine, serine, and threonine metabolism were identified. These pathways reveal that DR may be inextricably linked to levels of oxidative stress and inflammation in the organism. Finally, five active ingredients were identified as potential Q-markers of BHP against DR, namely ajugol, protocatechuic acid, tanshinone IIA, panaxatriol and puerarin. CONCLUSION: This study successfully clarified the efficacy and Q-markers of BHP through the Chinmedomics strategy, which is of great significance in determining the quality standards of BHP.

Effect of Chinese herbal compounds on ocular fundus signs and vision in conventional treated-persons with non-proliferative diabetic retinopathy: A systematic review and meta-analysis.

Background: Changes in fundus signs and loss of visual acuity are an important basis for screening and treating diabetic patients with retinopathy, and conventional Western medicine is moderately effective in treating diabetic retinopathy(DR),To systematically evaluate the effectiveness and safety of Chinese herbal compounds(CHCs) in the combined treatment of diabetic retinopathy. Method: Six electronic databases, including PubMed, were searched to screen eligible literature. Randomized controlled trials of non-proliferative diabetic retinopathy(NPDR) were included, in which the control group was treated with conventional Western-based drugs or retinal laser photocoagulation, and the intervention group was treated with CHCs in combination based on the control group.The Cochrane Risk of Bias Assessment Tool was used to evaluate the quality of the literature, and the RevMan 5.4 software was used for statistical analysis. Results: Compared with Conventional group alone,CHCs group was superior at improving clinical efficacy [RR=1.29, 95%CI=(1.23, 1.36),P<0.01] and best corrected visual acuity(BCVA) [MD=0.10,95%CI=(0.09,0.12),P<0.01],it was also superior at reducing the number of microangiomas [MD=-2.37, 95%CI=(-3.26, -1.49),P<0.01], microangioma volume [MD=-4.72, 95%CI=(-5.14, -4.29), P<0.01], hemorrhagic spots [MD=-2.05, 95%CI=(-2.51,-1.59), P<0.01], hemorrhagic area [MD=-0.76, 95%CI=(-1.06, -0.47), P<0.01], hard exudates [MD= -1.86, 95%CI=(-2.43, -1.28), P<0.01], cotton lint spots [MD= -0.93, 95%CI= (-1.31, -0.55), P<0.01], central macular thickness(CMT) [SMD=-1.52, 95%CI=(-1.85, -1.19),P<0.01], Chinese medicine evidence score [SMD=-1.33,95%CI=(-1.58, -1.08),P<0.01], fasting blood glucose (FBG) [SMD=-0.47, 95%CI=(-0.61,-0.33),P<0.01], 2h postprandial blood glucose(2hPBG) [SMD=-0.87, 95% CI=(-1.06, -0.67), P<0.01], glycosylated hemoglobin (HbAlc) [SMD=-0.76, 95%CI=(-1.16, -0.3),P<0.01], total cholesterol(TC) [SMD=-0.33,95%CI=(-0.51,-0.16),P<0.01],and CHCs group with less adverse events occurred [RR=0.46, 95%CI=(0.29, 0.74),P<0.01]. Conclusion: CHCs combined with conventional medicine for NPDR has better clinical efficacy and higher safety, but the above findings need further validation in more large sample, multicenter, and low-bias RCTs due to the limitation of the quality and quantity of included literature. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, identifier CRD42022342137.

Investigation on the Q-markers of Bushen Huoxue Prescriptions for DR treatment based on chemometric methods and spectrum-effect relationship.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic retinopathy (DR) is a kind of complex complication of late diabetes mellitus with high incidence and risk of blindness. Bushen Huoxue Prescription (BHP), which consists of Rehmanniae radix (RR), Salviae miltiorrhizae radix et rhizoma (SMRR), Ginseng radix et rhizome (GRR) and Puerariae lobatae radix (PLR), has an active effect on the treatment of DR. However, the quality markers (Q-markers) of BHP are not entirely clear. PURPOSE: This study aimed to screen the Q-markers of BHP for DR treatment based on the establishment of spectrum-effect relationship and verified experiment. MATERIALS AND METHODS: In this study, 12 BHP samples (S1-S12) for fingerprint analysis and pharmacological evaluation were prepared according to a four-factor and twelve-level uniform design. High performance liquid chromatography-ultraviolet detector-evaporative light scattering detector (HPLC-UV-ELSD) was employed to analyze the fingerprint on the basis of the characteristics of BHP components. The evaluation of sample similarity was carried out by similarity analysis (SA) and hierarchical cluster analysis (HCA). The pharmacological indicators, including expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) in the retina of Sprague Dawley (SD) rats induced by streptozotocin (STZ), were detected by enzyme-linked immunosorbent assay (ELISA). Besides, the spectrum-effect relationship between common peaks of fingerprints and the pharmacological results was investigated by partial least squares regression (PLSR) and canonical correlation analysis (CCA). The results of spectrum-effect relationship were verified by the expression of VEGF and HIF-1α on primary culture retinal Müller cells induced by hyperglycemia and hypoxia. RESULTS: In the HPLC-UV-ELSD fingerprint, 23 common peaks in UV and 14 common peaks in ELSD were identified. The pharmacological results indicated that the expression of VEGF and HIF-1α in the retina of SD rats was inhibited by 12 BHP samples to varying degrees compared with the model group. Based on SA and heatmap of HCA, S4 and S8 were clearly distinguished from other samples. The results of PLSR and CCA revealed that the contents of puerarin, daidzin, salvianolic acid B and ginsenoside Rb1 were inversely correlated with the expression of VEGF and HIF-1α. Hence, the four compounds may be the main active components to prevent and treat DR. The results of intervention on primary culture retinal Müller cells showed that puerarin, daidzin, salvianolic acid B, and ginsenoside Rb1 can significantly inhibit the expression of VEGF and HIF-1α. CONCLUSIONS: The spectrum-effect relationship of BHP was successfully established, and the Q-markers of BHP for the prevention and treatment of DR were preliminarily confirmed. It provides a feasible method for the research of quality control.

Efficiency of chitosan application against Phytophthora infestans and the activation of defence mechanisms in potato.

Late blight, caused by the oomycete Phytophthora infestans, is one of the most devastating diseases that results in huge losses of potato crops worldwide. Chitosan as a defence elicitor can induce plant innate immunity against pathogen infection, but the efficiency and specific defence mechanism of chitosan against late blight in potato have not been elaborated. In this study, we demonstrated that the application of chitosan significantly enhanced potato resistance and reduced P. infestans infection in potted potato and in the field. Large-scale transcriptomic analysis suggested that chitosan preferentially activated several important pathways related to the plant defence response. Notably, we revealed that chitosan triggered pattern-triggered immunity responses in potato. Chitosan could trigger pattern recognition receptors to initiate intracellular signalling, and gradually amplify the immune signal. qRT-PCR verification showed that chitosan induced the expression of defence-related genes in potato. Moreover, treatment with chitosan result in Induced Systemic Resistance (ISR) in potato, including an accumulation of plant hormone salicylic acid, increase in the level of phenylalanine ammonia lyase activity and a content decrease of malondialdehyde. These findings help elucidate chitosan-mediated activation of the immune system in potato and provide a potential ecofriendly strategy to control potato late blight in the field.

The effects of Bushen Yiqi Huoxue prescription and its disassembled prescriptions on a diabetic retinopathy model in Sprague Dawley rats.

BACKGROUND: Diabetic retinopathy (DR) is one of the most serious complications in the late stages of diabetes, with a complex mechanism. As a complication affecting local lesions, few studies have compared differences of cytokine expression in the serum and retina. Owing to the specific value of traditional Chinese medicine (TCM) to complex diseases, TCM research has recently boomed in the prevention and treatment of diabetes. Bushen Yiqi Huoxue (BYH) prescription is a Chinese herbal compound that has been independently developed by our research group and has been proved to have a positive effect on DR; however, its specific mechanism and compatibility rule remain to be further explored. OBJECTIVE: To construct a DR model of Sprague Dawley (SD) rats, simultaneously detect multiple factor expression in the serum and retina of rats, explore the effect of BYH prescription and its disassembled prescriptions on DR, and discuss the influence of various compatibility combinations. METHODS: BYH prescription was disassembled into two new compatibilities in the absence of Rehmanniae Radix (Yiqi Huoxue prescription, YH prescription) or Ginseng Radix et Rhizoma (Bushen Huoxue prescription, BH prescription). Male SD rats were induced using streptozotocin + high-fat and high-sugar diet to establish DR models and were divided into groups, then the intragastric administration and sampling. The body weight and fasting blood glucose of rats were continuously recorded during feeding; pathophysiological status observation of the retina by haematoxylin and eosin (HE) staining; advanced glycation end products (AGEs) and haemoglobin A1c (HbA1c) level detection in the rat serum by enzyme-linked immunosorbent assay; and the Luminex technique was used to detect the ICAM-1, IL-1ß, IL-6, TNF-α and vascular endothelial growth factor (VEGF) expression concentrations in the retinal tissue and serum. RESULTS: The results of blood glucose, body weight and HE staining proved that the model was successfully constructed, and the three combinations could reduce the retinal injury in DR rats. Serum AGEs and HbA1c levels of the model group increased compared with the control group (CG). Compared with the DR model group, only AGEs decreased in the BYH group, while the AGEs and HbA1c levels were significantly inhibited in the YH and BH groups, showing a significant correlation between the expression of AGEs and HbA1c in the serum of DR rats. In the serum of rats, IL-1ß, IL-6, TNF-α and VEGF concentrations in the DR model group increased, although no statistical difference was observed in the ICAM-1 data compared with the CG. Compared with the DR model group, the IL-1ß, IL-6 and TNF-α expression decreased in the BYH group. Moreover, the IL-6 and TNF-α expression decreased in the YH group and only the IL-6 expression decreased in the BH group. In the retina tissue, the model group had higher ICAM-1, IL-1ß, IL-6, TNF-α and VEGF levels than the CG. Compared with the DR model group, TNF-α in the BYH group rats decreased, and the ICAM-1, IL-6 and TNF-α concentrations decreased in the YH and BH groups. Furthermore, differences in the ICAM-1 and VEGF expression in the serum and retina existed. CONCLUSION: BYH compound and its disassembled prescriptions could improve the DR model rats induced with streptozotocin + high-fat and high-sugar diet, respectively, by inhibiting chronic blood glucose, AGEs, or inflammation response. The expression level and location of each factor are different, confirming that the effect of TCM prescriptions is not the simple addition of each single drug or its chemical components, but the rationality of its internal compatibility combination. Further, ICAM-1 and VEGF have exactly different expression levels, suggesting more attention to be paid by other researchers or doctors in future studies.

Urine metabolomics study of Bushen Huoxue Prescription on diabetic retinopathy rats by UPLC-Q-exactive Orbitrap-MS.

Diabetic retinopathy (DR) is a common microvascular complication of diabetes and remains one of the leading causes of blindness worldwide. Previous studies have shown that Bushen Huoxue Prescription (BP) possesses an effect on preventing and treating DR, but the mechanisms of action are not entirely clear. In order to clarify the mechanisms, the pharmacodynamical assessments of BP were investigated. Combining the pharmacodynamical studies, it was found that BP could inhibit high expression of VEGF and HIF-1α, indicating significant therapeutic effects on DR. In order to further investigate the mechanism of BP in treating DR, a urine metabolomics method based on ultrahigh-performance liquid chromatography coupled with Q-exactive quadrupole-electrostatic field Orbitrap mass spectrometry was established to observe the metabolic variations in DR rats and investigate the therapeutic effect of BP on DR. As a result, nine potential biomarkers associated with DR were found. The metabolic pathways related to these compounds were explored, and the results showed that these biomarkers were mainly associated with gut microbial metabolism, lipid metabolism and tryptophan metabolism. The results for the pharmacodynamics and metabolomics provided a theoretical basis for clarifying the mechanism of BP in the treatment of DR.

[Effect of Bushen Huoxue Compound on Retinal Müller Cells in High Glucose or AGEs Conditions].

OBJECTIVE: To explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method. METHODS: The retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Compared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05). CONCLUSIONS: AGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

[Effect of drug-serum with Chinese drugs for nourishing shen and activating blood on the glutamate release in purified retinal ganglion cells cultured in high-glucose conditions].

OBJECTIVE: To investigate the impacts of steady high-glucose or fluctuated glucose conditions on glutamate (Glu) release in purified retinal ganglion cells (RGCs) cultured in vitro, and the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on it. METHODS: RGCs of neonatal SD rats were cultured by antibody combined two-step purified method in different conditions: the simulated normal condition, the steady high-glucose condition and the fluctuated glucose condition, and they were intervened with S-NSAB. Thereby, the experiment was carried out in 6 groups, i.e. the normal control group (A), the S-NSAB intervened group (B), the steady high-glucose cultured group (C), the steady high-glucose cultured and S-NSAB intervened group (D), the fluctuated glucose cultured group (E), and the fluctuated glucose cultured and S-NSAB intervened group (F). Content of Glu in the extracellular fluid was detected at 24, 48 and 72 h after intervention with a full-automatic biochemical analyzer. And the data obtained were statistically analyzed with SPSS 13.0 soft ware. RESULTS: Release of Glu at 24 h after intervention in Group E (256.33 +/- 25.73 mg/L) was obviously higher than that in Group A and Group C (134.22 +/- 9.14 mg/L and 141. 17 +/- 22.13 mg/L, P < 0.05); at 24 h and 72 h in Group B (124.50 +/- 10.30 mg/L and 30. 17 +/- 2.97 mg/L) was obviously lower than in Group A respectively (P < 0.05); in Group D at 24 h (127.50 +/- 16.94 mg/L), 48 h (26.17 +/- 3.99 mg/L) and 72 h (27.67 +/- 3.49 mg/L) were lower than in Group C; in Group F at 24 h (228.33 +/- 18.41 mg/L) and 72 h (28.00 +/- 2.41 mg/L) were lower than in Group E respectively at the corresponding time points. CONCLUSIONS: Fluctuated glucose condition could obviously increase the Glu release of RGCs, to cause extracellular large amount Glu accumulation, which induces the exciting neurotoxicity to RGCs and finally to aggravate the injury on cells. S-NSAB could reduce the Glu release to some extent in the steady-high or fluctuated glucose conditions, diminish the injury of RGCs from exciting neurotoxicity of Glu, and it might be one of the intervening pathways of Chinese drugs for NSAB in preventing and treating DRP.

[Effect of serum contained Chinese drugs for nourishing Shen and activating blood on activity of purified retinal ganglion cells cultured in high glucose medium].

OBJECTIVE: To investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on activity of purified retinal ganglion cells (RGCs) cultured in high glucose medium. METHODS: Purified RGCs of SD rats were cultured in stimulative stable high glucose (50 mmol/L) condition (SHG) and fluctuated glucose condition (FGC) separately, they were intervened with S-NSAB, and the lactate dehydrogenase (LDH) leakage was detected by spectrophotometer for estimating the activity of RGCs. RESULTS: LDH leakage (U/L) in SHG culture was 1 349.17 +/- 215.50 at 24 h, 1220.24 +/- 124.53 at 48 h and 1982.14 +/- 219.03 at 72 h, all significantly lower than that in normal control at the corresponding time points (1628.10 +/-122.10, 1484.13 +/- 127.55 and 2155.75 +/- 140.44, respectively, P<0.05), whereas it was obviously higher in FGC culture at 72 h (2299.60 +/- 88.35), showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points (P<0.05). The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h (1797.62 +/- 146.40) and in FGC at 48 h (1259.92 +/- 87.74) and 72 h (1940.40 +/- 155.47), the difference between pre- and post-intervention was significant (P<0.05). CONCLUSION: Fluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells; S-NSAB could improve these abnormalities in either SHG or FGC condition, which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.