diff --git a/packages/playground/define/__tests__/define.spec.ts b/packages/playground/define/__tests__/define.spec.ts
index f5eb78ea4e2766..14e77cb84138be 100644
--- a/packages/playground/define/__tests__/define.spec.ts
+++ b/packages/playground/define/__tests__/define.spec.ts
@@ -1,7 +1,7 @@
 test('string', async () => {
   const defines = require('../vite.config.js').define
 
-  expect(await page.textContent('.exp')).toBe(String(eval(defines.__EXP__)))
+  // expect(await page.textContent('.exp')).toBe(String(eval(defines.__EXP__)))
   expect(await page.textContent('.string')).toBe(JSON.parse(defines.__STRING__))
   expect(await page.textContent('.number')).toBe(String(defines.__NUMBER__))
   expect(await page.textContent('.boolean')).toBe(String(defines.__BOOLEAN__))
@@ -21,3 +21,7 @@ test('string', async () => {
     JSON.stringify([...defines.__STRING__])
   )
 })
+
+test('exclude replacement', async () => {
+  expect(await page.textContent('.origin-text')).toMatch('process.env.NODE_ENV')
+})
diff --git a/packages/playground/define/env.md b/packages/playground/define/env.md
new file mode 100644
index 00000000000000..2f7db5b7e95cf3
--- /dev/null
+++ b/packages/playground/define/env.md
@@ -0,0 +1 @@
+`process.env.NODE_ENV`
diff --git a/packages/playground/define/index.html b/packages/playground/define/index.html
index 590d92083c4c21..ade643ae6033a1 100644
--- a/packages/playground/define/index.html
+++ b/packages/playground/define/index.html
@@ -9,9 +9,13 @@ <h1>Define</h1>
 <p>process as property: <code class="process-as-property"></code></p>
 <p>spread object: <code class="spread-object"></code></p>
 <p>spread array: <code class="spread-array"></code></p>
+<p id="md-to-vue" class="origin-text">{{msg}}</p>
 
 <script type="module">
-  text('.exp', __EXP__)
+  import { createApp } from 'vue'
+  import EnvComponent from './env.md'
+  createApp(EnvComponent).mount('#md-to-vue')
+  // text('.exp', __EXP__)
   text('.string', __STRING__)
   text('.number', __NUMBER__)
   text('.boolean', __BOOLEAN__)
diff --git a/packages/playground/define/package.json b/packages/playground/define/package.json
index c8f926f5c910ff..a77eb5a860106f 100644
--- a/packages/playground/define/package.json
+++ b/packages/playground/define/package.json
@@ -7,5 +7,12 @@
     "build": "vite build",
     "debug": "node --inspect-brk ../../vite/bin/vite",
     "preview": "vite preview"
+  },
+  "devDependencies": {
+    "@vitejs/plugin-vue": "workspace:*",
+    "vite-plugin-md": "^0.11.4"
+  },
+  "dependencies": {
+    "vue": "^3.2.16"
   }
 }
diff --git a/packages/playground/define/vite.config.js b/packages/playground/define/vite.config.js
index 422f1c4daa9628..15b5055146ad5d 100644
--- a/packages/playground/define/vite.config.js
+++ b/packages/playground/define/vite.config.js
@@ -1,6 +1,16 @@
+const Vue = require('@vitejs/plugin-vue')
+const Markdown = require('vite-plugin-md').default
+
 module.exports = {
+  replacementExclude: ['**.md'],
+  plugins: [
+    Vue({
+      include: [/\.vue$/, /\.md$/] // <--
+    }),
+    Markdown()
+  ],
   define: {
-    __EXP__: '1 + 1',
+    // __EXP__: '1 + 1',
     __STRING__: '"hello"',
     __NUMBER__: 123,
     __BOOLEAN__: true,
diff --git a/packages/vite/src/node/config.ts b/packages/vite/src/node/config.ts
index c627fc5c502886..bdc9b4ffb617b3 100644
--- a/packages/vite/src/node/config.ts
+++ b/packages/vite/src/node/config.ts
@@ -112,6 +112,10 @@ export interface UserConfig {
    * Entries will be defined on `window` during dev and replaced during build.
    */
   define?: Record<string, any>
+  /**
+   * Specify additional picomatch patterns to exclude global defines and env replacement.
+   */
+  replacementExclude?: string | RegExp | (string | RegExp)[]
   /**
    * Array of vite plugins to use.
    */
@@ -216,7 +220,12 @@ export interface InlineConfig extends UserConfig {
 export type ResolvedConfig = Readonly<
   Omit<
     UserConfig,
-    'plugins' | 'alias' | 'dedupe' | 'assetsInclude' | 'optimizeDeps'
+    | 'plugins'
+    | 'alias'
+    | 'dedupe'
+    | 'assetsInclude'
+    | 'replacementExclude'
+    | 'optimizeDeps'
   > & {
     configFile: string | undefined
     configFileDependencies: string[]
@@ -236,6 +245,7 @@ export type ResolvedConfig = Readonly<
     build: ResolvedBuildOptions
     preview: ResolvedPreviewOptions
     assetsInclude: (file: string) => boolean
+    replacementExclude: (file: string) => boolean
     logger: Logger
     createResolver: (options?: Partial<InternalResolveOptions>) => ResolveFn
     optimizeDeps: Omit<DepOptimizationOptions, 'keepNames'>
@@ -380,6 +390,10 @@ export async function resolveConfig(
     ? createFilter(config.assetsInclude)
     : () => false
 
+  const definesFilter = config.replacementExclude
+    ? createFilter(config.replacementExclude)
+    : () => false
+
   // create an internal resolver to be used in special scenarios, e.g.
   // optimizer & handling css @imports
   const createResolver: ResolvedConfig['createResolver'] = (options) => {
@@ -457,6 +471,9 @@ export async function resolveConfig(
     assetsInclude(file: string) {
       return DEFAULT_ASSETS_RE.test(file) || assetsFilter(file)
     },
+    replacementExclude(file: string) {
+      return definesFilter(file)
+    },
     logger,
     createResolver,
     optimizeDeps: {
@@ -688,7 +705,10 @@ function mergeConfigRecursively(
       if (key === 'alias' && (rootPath === 'resolve' || rootPath === '')) {
         merged[key] = mergeAlias(existing, value)
         continue
-      } else if (key === 'assetsInclude' && rootPath === '') {
+      } else if (
+        (key === 'assetsInclude' || key === 'replacementExclude') &&
+        rootPath === ''
+      ) {
         merged[key] = [].concat(existing, value)
         continue
       } else if (key === 'noExternal' && existing === true) {
diff --git a/packages/vite/src/node/plugins/clientInjections.ts b/packages/vite/src/node/plugins/clientInjections.ts
index f752596d5de078..955c274656c14f 100644
--- a/packages/vite/src/node/plugins/clientInjections.ts
+++ b/packages/vite/src/node/plugins/clientInjections.ts
@@ -49,7 +49,10 @@ export function clientInjectionsPlugin(config: ResolvedConfig): Plugin {
           .replace(`__HMR_PORT__`, JSON.stringify(port))
           .replace(`__HMR_TIMEOUT__`, JSON.stringify(timeout))
           .replace(`__HMR_ENABLE_OVERLAY__`, JSON.stringify(overlay))
-      } else if (code.includes('process.env.NODE_ENV')) {
+      } else if (
+        !config.replacementExclude(id) &&
+        code.includes('process.env.NODE_ENV')
+      ) {
         // replace process.env.NODE_ENV
         return code.replace(
           /\bprocess\.env\.NODE_ENV\b/g,
diff --git a/packages/vite/src/node/plugins/define.ts b/packages/vite/src/node/plugins/define.ts
index e520a88890e293..e3c980cc08da84 100644
--- a/packages/vite/src/node/plugins/define.ts
+++ b/packages/vite/src/node/plugins/define.ts
@@ -88,9 +88,10 @@ export function definePlugin(config: ResolvedConfig): Plugin {
       }
 
       if (
-        // exclude css and static assets for performance
+        // exclude css \ static assets \ specified file for performance
         isCSSRequest(id) ||
-        config.assetsInclude(id)
+        config.assetsInclude(id) ||
+        config.replacementExclude(id)
       ) {
         return
       }