Basic medical research on traditional Chinese medicine (TCM) ophthalmology has made great progress in recent years. This mainly includes the following aspects: the gradual increase in the number and quality of relevant documents; the increase in research institutions and research specialists engaged in TCM ophthalmology-related basic medical research; and the focus of this research, mainly concentrated on dominant diseases in TCM ophthalmology. However, there are also some challenges. These are mainly reflected in the following aspects: the difficulty of transforming scientific research achievement; the lack of rigorous experimental design and in-depth research on the mechanisms of Chinese medicine; the lack of optimal utilization and integration of resources for research; and the unbalanced development of scientific research levels and resources. To a certain extent, these have hindered the development of basic medical research on TCM ophthalmology. In the future, we need to integrate more superior resources into the dominant diseases and key areas of TCM ophthalmology, and persist in the in-depth research of them; and insist on collaborative innovation and successful cooperation in order to better serve clinical medicine. This is the fundamental starting point and emerging direction for basic medical research on TCM ophthalmology.
Objective: To determine the neuroprotective effects of salidroside (Sal) on murine retinal ganglion cell (RGC) apoptosisinduced by N-methyl-D-aspartate (NMDA). Methods: Thirty-six C57BL6J mice were randomized into a blank normal control group, a model control group, and four experimental group (n=6 each). The right eyes of the experimental groups received a 1.5 μl intravitreal injection 0.1, 0.4, 2, or 8 mmol/L Sal (n=6 mice for each dose) 48 hours before NMDA was injected, and phosphate-buffered saline (PBS, 1.5 μl) were injected intravitreally in the model control group 48 hours before NMDA was injected. Intravitreal NMDA injections (1.5 μl at 0.1 mmol/L), were then performed in the model control group and in the experimental groups. PBS (1.5 μl) was injected intravitreally in the blank control group. Retinas derived from the mice were flatmounted and stained with hematoxylin and eosin to identify the RGCs. The number of RGCs was compared between NMDA and PBS-injected eyes for all groups. The levels of caspase-3 and caspase-8 were analyzed by Western Blot. Data were analyzed using ANOVA.Results: Compared with the blank model group, NMDA induced a significant increase of RGC apoptosis in the model control group and in the experimental groups. Compared with the model control group, Sal significantly inhibited RGC apoptosis in a dose-dependent manner (F=212.0, P<0.001), and the protein expression of caspase-3 and caspase-8 was significantly down regulated (F=168.3, P<0.001). Conclusions: Sal inhibited NMDA-induced RGC apoptosis and provided some neuroprotective effect for glutamatemediated RGC excitotoxicity.
Objective: To observe and explore the protective effects of breviscapine (BVP) on tert-butyl hydroperoxide (tBHP)-induced primary retinal ganglion cell (PRGC) apoptosis. Methods: In this experimental study,PRGCs were isolated by immunopanning from retinas of 1-day-old mice and maintained in culture for 3 days. To induce apoptosis, the PRGCs were then exposed to 0, 50, 100, or 200 μmol/L tBHP (100 μmol/L was chosen to establish models) in the absence or presence of 0, 10, 20, or 50 μmol/L BVP (20 μmol/L was chosen for follow experiments). The PRGCs were identified by immunofluorescence. Expression of Bcl-2, caspase 3, and synaptophysin were assessed by western blotting. Cell apoptosis was detected by the terminal uridine nick-end labeling (TUNEL) assay. Data analysis was performed by t-tests. Results: Compared with the blank control group, the proliferation of PRGCs was significantly inhibited by tBHP (P<0.05).Cell apoptosis increased (P<0.01), and the expressions of Bcl-2 and synaptophysin proteins were significantly down-regulated (P<0.05, P<0.01 respectively). Compared with the tBHP alone, the survival of PRGCs was increased by co-incubation of tBHP with BVP (P<0.01). Cell apoptosis was reduced, along with expressions of Bax (P>0.05) and cleaved-caspase-3 (P<0.001). The protein expressions of Bcl-2 and synaptophysin increased (P<0.05). The protective effects of BVP on tBHP-induced PRGC apoptosis were achieved in a concentration dependent manner. Conclusions: BVP can inhibit tBHP-induced PRGC apoptosis.
Objective: To study the effectiveness of a traditional Chinese medicine (TCM) used for soothing the liver and unblocking the orifices to determine if it can protect the optic nerve from high intraocular pressure in glaucoma by observing retinal ganglion cell (RGCs) counts and the ultrastructure and morphological changes of the retina and optic nerve in rats; to develop an effective TCM prescription to protect the optic nerve from glaucoma. Methods: In this experimental study, the 4th formula TCM prescriptions for unblocking the orifices to improve vision is based on the therapeutic principles of soothing the liver and unblocking the orifices. In this research study, 90 SD rats were used as experimental animals. Chronic high intraocular pressure (IOP) was established in rat models by injecting carbomer solution into the anterior chamber of the right eyes. The models were first divided into 3 groups based on different IOP time frame windows (1 week, 2 weeks, 3 weeks). Then each IOP group was divided into a model group (non-treated, 5 rats), negative control group (5 rats), positive control group (treated with a neurotrophic solution, 5 rats), low-dose treatment group (treated with 10 gkg-1 d-1 4th formula for blocking orifices, 5 rats), middle-dose treatment group (20 gkg-1 d-1, 5 rats) and high-dose treatment group (40 gkg-1 d-1, 5 rats). The rats in the treatment groups were given TCM prescriptions by intragastric administration. RGC counts were observed by a CMIAS series digital medical image analysis system. The ultrastructure of the retina and optic nerve was observed by electron microscopy. The data were analyzed by one-way ANOVA and LSD methods. Results: 1. RGC counts gradually reduced along with the duration of high intraocular pressure (F=87.67, 29.69, 33.38, 38.03, 33.67, 23.36, all P<0.001). Comparing the high- and middle-dose treatment groups with the positive and negative control groups (IOP at 1 week, 2 weeks, 3 weeks), the survival of RGCs significantly increased with the oral administration of the 4th formula for unblocking the orifices to improve vision and the differences were statistically significant (P<0.001). 2. In the model and negative control groups, the retinal structures were disordered, the retinal membrane was thinning, the mitochondria showed cavitation degeneration and cellular atrophy and necrosis were observed. In contrast, in every treatment group, disorder of the retinal structures was alleviated, the thickness of the retinal membrane slightly increased, and cavitation degeneration and cell atrophy were relieved. 3. In model and negative control groups, the arrangement of optic nerve axons was disordered, the density of axons was reduced, microfilament was dissolved, the mitochondria showed cavitation degeneration, organelles were swelled and damaged and the myelin sheath had degenerated. In contrast, in every treatment group, the edema of the myelin sheath of the optic nerve and mitochondria were relieved and medullary cord degeneration was repaired. Conclusions: The 4th formula for unblocking orifices to improve vision can improve the survival microenvironment of RGCs in rat models with high intraocular pressure, protect undamaged cells, repair mildly damaged RGCs, delay or prevent the descending changes of some damaged cells and reduce the apoptosis of RGCs. Therefore, the 4th formula for unblocking orifices to improve vision has protective effects on optic nerve damage from glaucoma.
Objective: To measure recruitment of bone marrow-derived mesenchymal stem cells from peripheral blood to the retina of royal college of surgeon (RCS) rats and the expression of ciliary neurotrophic factor (CNTF). We also investigated the mechanism by which Bu Shen Yi Jing Fang (BSYJ) could treat retinal degenerative diseases. Methods: This was an experimental study. RCS rats were injected with 1×107 green fluorescent protein- (GFP-) labeled bone marrow-derived mesenchymal stem cells (GFP-MSCs). The injected rats were assigned to a distilled water group or a BSYJ group according to selection from a random numbertable, and 39 rats in each group. Each rat in the respective groups was gavaged with either distilled water or BSYJ solution one day after injection. Normal SD rats were treated routinely as the control group. At 7, 14, and 28 days after treatment, retina flat mounts were prepared to assess the recruitment of GFP-MSCs to the retina. Quantitative polymerase chain reaction (Q-PCR) and immunofluorescence double labeling were used to determine the effect of BSYJ on expression of ciliary neurotrophic factor (CNTF) in the retina. Apoptosis of retinal cells was measured by the TUNEL method. Data were analyzed using ANOVA. Results: At 7, 14, and 28 days after treatment, the numbers of GFP-MSCs in the distilled water group were less than in the BSYJ group, and the difference was significant at 28 days (t =2.71, P=0.001). At 7, 14, and 28 days after treatment, CNTF mRNA expression in the BSYJ group was significantly higher than in the distilled water group (F=5.763, P=0.003; F=3.165, P=0.042; F=4.839, P=0.004). At 7 and 28 days after treatment, the apoptosis rate of retinal cells in BSYJ group was significantly less than in the distilled water group (F=0.002, P=0.001; F=0.307, P=0.004). Conclusions: BSYJ improved recruitment of peripheral blood stem cells to the retina and increased CNTF expression. The recruitment of stem cells by BSYJ may delay progression of retinal cell apoptosis and degeneration in RCS rats.
Objective: To study the molecular mechanism of LongdanXiegan Tang for treating uveitis. Methods:This was a experimntal study. The Chinese herbal medicine pharmacology data and analysis platform were used to search and to screen the effective components of a compound injection of LongdanXiegan Tang, and to analyze its possible therapeutic targets based on the network topology. By searching a variety of known disease target databases, we can screen the therapeutic target of uveitis disease and construct the active ingredient-disease treatment target network with the target of effective component treatment and to screen out the key nodes. Using topological analysis of the key nodes involved in the signalpathways and enrichment analysis, and screening out the possible Chinese medicine compound Radix LongdanXiegan Tang for the treatment of uveitis, may help to identify the signal pathways. Results: After the main active ingredient for screening and target prediction of LongdanXiegan Tang were completed, the therapeutic target of uveitis was screened, and an enrichment analysis of the signal pathways was completed. LongdanXiegan Tang can reduce the production of inflammatory cytokines and infiltration and affect the immune status, regulating the occurrence and development of uveitis by mainly regulating the MAPK signaling pathway and promoting both the GnRH and estrogen signaling pathways. Conclusions: LongdanXiegan Tang can treat uveitis primarily through the MAPK, GnRH and estrogen signaling pathways, regulating the inflammatory cells and inflammatory cytokines and autoimmunity.
Objective: To research the regulating effects of Xiaomengling on the tumor necrosis factor α (TNF-α), Janus kinase 2/signal transducer and activator of transcription 5 (JAK2/STAT5) signaling pathway, and to study its mechanism in preventing and curing diabetic retinal microangiopathy. Methods: This was an experimental research study. Fifty male SD rats were chosen and acclimated for 5 days. Eight rats were randomly chosen as the blank group; diabetic models were established in the other 42 rats with streptozotozin (STZ). Forty rats were modeled successfully and were divided randomly into 4 groups: 10 rats were the model group and 30 rats were the treatment group. In the treatment group, 10 rats were given a 0.1 g/100 g dose of Xiaomengling (low-dose group), 10 were given a 0.2 g/100 g dose of Xiaomengling (medium-dose group), and 10 were given a 0.3 g/100 g dose of Xiaomengling (high-dose group). After 4 months of treatment, body mass and blood sugar change were observed, and changes in the retinal ultrastructure were observed under light microscopy and electron microscopy. The protein expression change in TNF-α, JAK2/STAT5 was observed by Western blot. Relevant data were analyzed with repeated measures analysis of variance. Results: TNF-α, JAK/STAT expression was visible in the retinas of the blank group. More expression was visible in the model group; expressions were reduced successively in the low-dose, medium-dose and high-dose groups. The comparison between groups was statistically significantly (F=4.61, 4.23, 3.28, P<0.05). By observing retinal structures of rats under light microscopy, rats in the blank group had normal retinal structures. Rats in the model group had deranged distribution in each layer, obvious retinal edema and telangiectasia, decreased number of ganglion cells, karyopyknosis and inner nuclear layer vacuolar degeneration. Rats in the high-dose treatment group had default arrangements in each retinal layer, slight edema, a decreased number of ganglion cells and inner nuclear layers. Under retinal electron microscopy, rats in the blank group had normal retinal structures. The middle parts of the retinal rod outer segments of rats in the model group were ruptured, dissolved and degenerated. The mitochondrial cristae disappeared, and had no orderly pattern, their cones were irregular in shape, chromatin was distributed unevenly in the nucleus, the retinal capillary basement membrane was thickening and the channel was narrow. The spaces between the retinal rod outer segments in the high-dose treatment group were expanded and were not regular enough in alignment. Some of the mitochondria were swollen and deformed, the nuclear chromosomes in the cone cells had no orderly pattern, and the thickness of the retinal capillary basement membrane was uneven. Conclusions: Xiaomengling has significant regulating effects on the TNF-α and JAK2/STAT5 signal transduction pathway of diabetic rats, and can effectively slow or prevent diabetic retinal microangiopathy.
Objective: To study the effect of Yin Xue deficiency and Yang Qi deficiency Syndrome on oxygen saturation of retinal vessels in eyes of patients with chronic nephropathy and diabetes. Methods: This case control study, which was based on traditional Chinese medicine (TCM) syndrome elements research, included 64 cases of type 2 diabetes, 83 cases of chronic nephropathy, and 103 normal subjects in Sichuan Provincial Hospital of Traditional Chinese Medicine from November 2014 to January 2016. Average blood oxygen saturation was measured in the retinal artery and vein, and the differences between them were calculated. Differences in retinal arterial and venous oxygen saturation levels associated with the TCM syndromes Yin Xue and Ying Qi deficiencies were analyzed using variance analysis and Kruskal-Wallis correlation. Results: Patients with Yin Xue deficiency and Yang Qi deficiency had similar oxygen saturation in the retinal artery (101.4% ± 10.2% and 98.3% ± 6.0% respectively, P > 0.05). In both groups, the oxygen saturation was significantly higher than in normal subjects (H=15.74, P < 0.001). The oxygen saturation in the retinal veins of patients with Yin Xue and Yang Qi deficiency were similar (62.2% ± 8.4% and 63.2% ± 5.7% respectively), and both similar to normal subjects (F=0.56, P=0.569). For Yin Xue deficiency patients, the arteriovenous difference in oxygen saturation, 39.2% ± 7.8%, was higher than for Yang Qi deficiency patients, 35.0% ± 5.8% (P < 0.001). Conclusions: In Yin Xue deficiency patients, the arteriovenous difference in blood oxygen saturation is higher than in Yang Qi deficiency patients. This suggests that patients with Yin Xue deficiency have a higher rate of oxygen consumption than Yang Qi patients.
Objective: We investigated the in vivo and in vitro effects of microRNA-182 (miR-182) , which modulates post-transcriptional regulation of gene expression, on corneal epithelial cell proliferation, migration, and wound healing. Methods: In this experimental research, mouse corneal epithelial wound healing of five mouse was initiated by a scrape injury. Realtime reverse transcriptase polymerase chain reaction (RT-PCR) was performed to detect the expression level of miR-182 in the mouse corneal epithelium during the wound healing process (48 hours post scraping). Lipofectamine RNAiMAX reagent was used to transfect human corneal epithelial cells (HCECs) in vitro with either miR-182 or negative control RNA oligo. Cell proliferation assay (MTS), cell colony-forming assay, and flow cytometry were used to evaluate the effects of miR-182 on cell proliferation and cell cycle. An in vitro wound-healing assay (the HCECs were wounded by placing a scratch along the surface of each culture) was carried out to evaluate the effect of miR-182 on cell migration. Data were analyzed using independent t test. Results: The in vivo expression of miR-182 in experimentally wounded corneal epithelia was significantly lower than in the non-wounded controls (t=147.6, 79.2, 136.8, 41.3, 89.8, all P<0.001). In vitro, the relative number of cells in the experimental group transfected with miR-182 was (81±5)%, which was significantly less than that in the control group transfected with negative control RNA oligo, 100% (t=6.6, P=0.003). Further, the number of clones in the experimental group was remarkably decreased compared to that in the control group. The percentage of transfected cells in G1 arrest was (49±7)%, which was significantly higher than that in the control group, (35±4)% (t=-3.0, P=0.041). In addition, the migration distance of HCECs in the transfected cells, 99±12 μm, was shorter than in the control cells, 213±14 μm (t=10.8, P=0.001). Conclusions: miR-182 negatively regulates the corneal epithelial wound healing process by inhibiting corneal epithelial cell proliferation and migration.
More and more patients with ametropia are wearing contact lenses (CLs) to correct refractive errors.According to statistics, globally there are 140 million CL wearers. At present, the evaluation of CL safety is mainly based on the material and the wearer's own factors. However, for different wearers working in different industries and environment, the safety hazards are often unknown. The environment of the CL wearer has becoming an indispensable factor to evaluate the safety of CL wear. However, there is currently no consensus on the safety of wearing CLs in different environments. Therefore, this review aimed to achieve a more comprehensive understanding of the impact of different environments on CL wearers to achieve the goal of wearing CLs safely.
