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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (10): 1140-1148.doi: 10.3736/jcim20121011

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Effects of Chinese herbal medicine Shenxiong Yujing Granule on regeneration of neural cells in rats with diabetes-associated cerebral ischemia

Hui Han1,2, Li-min Wu3, Ming-xiang Han2(), Wen-ming Yang1, Dao-jun Xie1, Zhao-hui Fang4, Dan-chen Chu1, Fang Fang1   

  1. 1. Department of Neurology, the First Affiliated Hospital, Anhui University of Traditional Chinese Medicine, Hefei 230031, Anhui Province, China
    2. Famous Experts in Traditional Chinese Medicine Workroom of State Administration of Traditional Chinese Medicine, the First Affiliated Hospital, Anhui University of Traditional Chinese Medicine, Hefei 230031, Anhui Province, China
    3. Medical Research Center, Anhui Provincial Hospital, Hefei 230001, Anhui Province, China
    4. Department of Endocrinology, the First Affiliated Hospital, Anhui University of Traditional Chinese Medicine, Hefei 230031, Anhui Province, China
  • Received:2012-03-30 Accepted:2012-04-20 Online:2012-10-20 Published:2018-10-15
  • Contact: Han Ming-xiang E-mail:hmx401125@sina.com

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Objective: To establish a rat model of diabetes-associated cerebral ischemia due to qi and yin deficiency and blood stasis, and to investigate the effects of Radix Ginseng, Rhizoma Chuanxiong, Rhizoma Polygonati Odoratiand Rhizoma Polygonati Sibirici Granule (Shenxiong Yujing Granule), which has the function of strengthening qi, nourishing yin, and activating blood, on proliferation, differentiation and survival of neural cells in rats with diabetes-associated cerebral ischemia. .

Methods: Rats were divided into sham-operation, diabetes plus ischemia reperfusion injury model, Shenxiong Yujing Granule and Radix Ginseng and Rhizoma Chuanxiong Granule (Shenxiong Granule) groups with 20 rats in each. The 5-bromo-2′-deoxyuridine (BrdU) incorporation assay and immunohistochemical method were used to investigate the proliferation, differentiation and survival of neural cells in dentate gyrus of rats with diabetes-associated cerebral ischemia.

Results: The number of newly proliferating cells in subgranular zone of dentate gyrus was increased in the model group, but there was no significant difference compared with 7 day treatment with Shenxiong Yujing Granule. Shenxiong Yujing Granule significantly increased the survival rate and promoted the differentiation of newly proliferating neurons after 21-day treatment (P<0.01). In addition, the beneficial effect of Shenxiong Yujing Granule was considerably greater than that of the Shenxiong Granule (P<0.01).

Conclusion: Shenxiong Yujing Granule can increase the survival rate and promote the differentiation of newly proliferating neurons in rats with diabetes-associated cerebral ischemia of dual deficiency of qi and yin and blood stasis obstructing the collaterals. The effect is greater than that of Shenxiong Granule.

Key words: drugs, Chinese herbal, diabetes mellitus, brain ischemia, nerve regeneration, neurons, disease models, animal, astrocytes, rats

Figure 1

Effects of Shenxiong Yujing Granule on neurocyte proliferation in dentate gyrus of hippocampus in rats with diabetes and ischemia reperfusion injury (immunohistochemical method, fluorescence microscopy) Shenxiong Yujing Granule is composed of Radix Ginseng, Rhizoma Chuanxiong, Rhizoma Polygonati Odorati and Rhizoma Polygonati Sibirici Granule and Shenxiong Granule is composed of Radix Ginseng and Rhizoma Chuanxiong. The sections were labeled with BrdU antibody (blue, nuclear staining). A to D showed BrdU-positive neurocytes on day 7. E to H showed BrdU-positive neurocytes on day 21. A and E: Sham-operation group; B and F, Diabetes plus ischemia reperfusion injury group; C and G: Shenxiong Yujing Granule group; D and H: Shenxiong Granule group. The black arrows in pictures point to the BrdU-labeled cells. Bar: 50 μm; BrdU: 5-bromo-2′-deoxyuridine."

Figure 2

Quantitative analysis of the effects of Shenxiong Yujing Granule on neurocyte survival and proliferation in dentate gyrus of hippocampus in rats with diabetes and ischemia reperfusion injuryShenxiong Yujing Granule is composed of Radix Ginseng, Rhizoma Chuanxiong, Rhizoma Polygonati Odorati and Rhizoma Polygonati Sibirici Granule and Shenxiong Granule is composed of Radix Ginseng and Rhizoma Chuanxiong. A: BrdU-positive neurocytes on the 7th and 21st days after cerebral ischemia; B: Ratio of BrdU-positive neurocytes on the 7th and 21st days, namely the ratio of new regenerative and surviving cells. **P<0.01, vs sham-operation group; △P<0.05, △△P<0.01, vs diabetes plus ischemia reperfusion injury group; ▲P<0.05, vs Shenxiong Granule group."

Figure 3

Effects of Shenxiong Yujing Granule on neurocyte and astrocyte differentiation in dentate gyrus of hippocampus in rats with diabetes and ischemia reperfusion injury on day 21 (immunofluorescence method, fluorescence microscopy) The sections were isolated from a rat in the Shenxiong Yujing Granule group and were double-labeled with NeuN (A, green, nuclear staining) and BrdU (B, red, nuclear staining) or with GFAP (D, green, cytoplasmic staining) and BrdU (E, red, (blue, nuclear staining)) antibody. Some BrdU-positive cells colocalized with NeuN-positive cells (C, yellow); some BrdU-positive cells colocalized with GFAP-positive cells (F). The white arrows in pictures point to NeuN- or GFAP-BrdU double-labeled cells. Bar: 20 μm. The quantitative analysis of neuronal differentiation of new-born cells was shown in G. The percentage is recorded as the number of double-positive cells divided by that of BrdU-positive cells (G). BrdU: 5-bromo-2′-deoxyuridine; NeuN: neuron-specific nuclear protein; GFAP: glial fibrillary acidic protein. **P<0.01, vs sham-operation group; △△P<0.01, vs diabetes plus ischemia reperfusion injury group; ▲▲P<0.01, vs Shenxiong Granule group."

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